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Prolactin (PRL) has long been known to be a hormone responsible for mammary gland development and lactation in females, whereas its role in males is still unclear. Thus, we investigated male mouse (m) PRL protein and mRNA expression in spermatozoa at various differentiation stages in the testes. Quantitative RT-PCR and in situ hybridization detected the expression of PRL not only in Leydig cells but also in germ cells, in particular in spermatogonia. The nucleotide sequence of testis PRL mRNA was the same as that in the pituitary. The mPRL was detected in Leydig cells and in round and elongated spermatids of the testes by immunohistochemistry. Immunoblotting detected 2 forms of mPRL in the testes, one form was 23-kDa PRL, and the other form was smaller than full-length PRL. Based on these results, we focused on N-terminal cleaved PRL to determine its involvement in spermatogenesis. Immunohistochemistry using two sets of antibodies, one that recognized full-length PRL and N-terminal cleaved PRL and another that recognized full-length PRL and C-terminal cleaved PRL, suggested that intact PRL was localized in the nucleus of round spermatids, while N-terminal cleaved PRL variants were localized in the Golgi apparatus of the sperrmatid nuclei of round spermatids, cytoplasms of elongated spermatids and in the spermatozoa tails. These findings suggest that PRL is ectopically expressed in the spermiogenesis and spermatogenesis and that cleaved PRL variants were localized in the Golgi apparatus of spermatids and in spermatozoa tails.

Study Design: A prospective study on the clinical outcomes in patients with tuberculous spondylitis treated by a 2-stage operation (posterior and anterior) using posterior spinal instrumentation., Objective: To evaluate the clinical outcomes of the 2-stage surgical treatment (first stage: placement of posterior instrumentation and second stage: anterior debridement and bone grafting) for tuberculous spondylitis., Summary of Background Data: There have been few reports describing the effects of 2-stage surgical treatment for tuberculous spondylitis., Methods: Ten patients (5 men and 5 women) with tuberculous spondylitis were treated by 2-stage operations. Age at the initial operation was 64.6+/-14.8 years (average+/-SD) (range: 47 to 83 y). The clinical outcomes were evaluated before and after the surgery in terms of hematologic examination, pain level, and neurologic status. Bone fusion and changes in sagittal alignment were examined radiographically., Results: All patients showed suppression of infection, bony fusion, relief of pain, and recovery of neurologic function. No significant changes were observed in kyphosis angle at the final follow-up. There were no incidences of severe complications or recurrence., Conclusions: Our results showed that posterior and anterior 2-stage surgical treatment for tuberculous spondylitis is a viable surgical option for cases in which conservative treatment has failed. However, the changes in sagittal alignment showed that this strategy provides limited kyphosis correction.

This study examined the prevalence of degenerative changes of knee menisci in aging and evaluated the diagnostic values of magnetic resonance (MR) imaging for assessing meniscal pathology in an older population. Eighty-five knees of asymptomatic volunteers over the age of 40 were scanned using MR imaging. Meniscal abnormalities were graded from 0 to 3 according to intrameniscal MR signals. The subjects were divided into two groups based on the presence or absence of radiographic osteoarthritis. Group I included 43 knees that had normal radiographs and group II consisted of 42 knees that had radiographic evidence of osteoarthritis. Degenerative changes in the menisci involved primarily the posterior segment of the medial meniscus in both groups. Signal changes in the other segments were of a significantly lesser grade than that in the posterior segment of the medial meniscus. The meniscal grade in each segment was significantly higher in group II than in group I. In group I, only two menisci (4.6% ) showed grade 3 signals, even in the posterior portion of the medial meniscus, compared to 21 (50.0%) in group II. Frequency of asymptomatic grade 3 was relatively low even in older subjects if there was no evidence of radiographic osteoarthritic changes. Abnormal MR signals are more likely to have clinical significance, in patients with radiographic changes on plain X-ray.

Introduction: A case of combined epiconus and cauda equina syndrome due to multilevel spinal canal stenosis of the thoracolumbar spine is reported., Methods: A 76-year-old man with multilevel spinal canal stenosis of the thoracolumbar spine (Th11-12, L2-S) who showed symptoms of epiconus syndrome was reported. First, we performed anterior decompression and fusion at the thoracolumbar junction (decompression: Th11-12, fusion: Th10-L2), which ameliorated his symptom partially. However, he presented cauda equina symptoms. Then, he underwent posterior spinal decompression (L3-5) and fusion (Th12-L5)., Results: After anterior decompression, several symptoms disappeared. However, motor and sensory disturbance below L4 and bladder-bowel disturbance remained. We then performed a secondary operation. At three years' follow-up, he was able to walk with the aid of a cane., Conclusions: Combined epiconus and cauda equina syndrome due to multilevel spinal canal stenosis was treated by combined two-stage anterior and posterior decompression. In this case, multilevel decompression via anterior and posterior approaches was necessary to relieve the symptoms.

Lactation-associated expression of GD1 alpha ganglioside in murine mammary glands was found to be due to the increasing specific activities of Gg4Cer alpha2,3- and GM1b alpha2,6-sialyltransferases in the glands from 12th day of gestation. The gene for GM1b alpha2,6-sialyltransferase, mST6GalNAcV, which was not detected in nonpregnant glands, appeared at 12th day of gestation and increased in the following gestational and lactation periods. At 3rd day of lactation, the amounts of lipid-bound sialic acid (LSA) in the mammary glands and milk of HR-1 mice were 99.3 +/- 8.5 microg per gram of dried tissue and 2.9 microg per ml, GD1 alpha comprising 64.0% and 80.5% of the total LSA, respectively, and GD1 alpha in milk was found to be preferentially distributed in the fat globule fraction. When the mammary epithelial cells at 15th day of gestation were cultured in prolactin- and epidermal growth factor (EGF)-containing media, the synthesis of fat globules and casein, together with the enhanced synthesis of GD1 alpha, were observed in the cells in prolactin medium, indicating that synthesis of GD1 alpha occurs in association with milk production as a prolactin-dependent event. Thus, GD1 alpha ganglioside, which is characteristically distributed in the cerebellar Purkinje cells of the murine brain, is supplied to neonates through the milk of the mother.

Phosphorylated prolactin (PPRL) is considered to be the most quantitatively important post-translationally modified form of prolactin (PRL) in rodents. We recently detected two different types of PPRL in the mouse pituitary gland; one was phosphorylated at serine and the other was phosphorylated at serine/threonine. Furthermore, we showed that there are obvious differences in the ratios between PPRLs and non-phosphorylated PRL in the pituitary gland based on age and sex and that estrogen influences PRL phosphorylation at serine in female mice. In the present study, we examined whether estradiol (E2) increases serine PPRL in the male pituitary gland in the same manner as in the female pituitary gland and examined whether PPRL is released into serum. We first determined the relative amounts of intrapituitary PPRLs in male mice under different pharmacological conditions that increased PRL secretion. The results indicated that treatment with E2 increases serine PPRL. We then performed two-dimensional electrophoresis and immunoblotting analysis after immunoprecipitation with anti-mouse PRL antibody using male and female sera under different pharmacological conditions that increased PRL secretion. The results of this experiment indicated that there were PRLs phosphorylated at serine and serine/threonine in the female serum but not in the male serum. The levels of PPRLs in sera were greatly increased with the E2 treatment for both male and female sera. Furthermore, we examined the effect of E2 on PPRL synthesis in cultured male pituitary glands. In this experiment, we observed increased serine PPRL synthesis and stronger immunohistochemical staining of PRL cells with E2 treatment. These findings suggested that serine PPRL synthesis and secretion were influenced by estrogen.

In this study, LAMP markers linked to shelf-life in melon (Cucumis melo L.) were developed by converting a cleaved amplified polymorphic sequences (CAPS) marker (C2). The CAPS-PCR fragments from the long-shelf-life melon (O-3) and short-shelf-life melon (Nat-2) were cloned and sequenced to construct LAMP primers. A single nucleotide polymorphism (SNP) was identified between O-3 and Nat-2. LAMP primers were designed to detect the SNP. In the LAMP reaction to detect long-shelf-life melon, the turbidity of the templates using O-3, F1, homozygous long-shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. In contrast, the turbidity of Nat-2 and homozygous short-shelf-life F2 lines did not increase even after 90 min. In the LAMP reaction to detect short-shelf-life melon, the turbidity of the templates using Nat-2, F1, homozygous short-shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. But the turbidity of O-3 and homozygous long-shelf-life F2 lines did not increase after 90 min. This attests to the high reliability and usefulness of LAMP for marker-assisted selection.

An immunocapture reverse transcription loop-mediated isothermal amplification (IC/RT-LAMP) was developed for the detection of tomato spotted wilt virus (TSWV) from chrysanthemum. This method enabled sensitive, reproducible and specific detection of TSWV from chrysanthemum plants. In the RT-LAMP method, TSWV genomic RNA could be amplified under isothermal (65 degrees C) conditions within 1 h. The resulting amplicons were detected by the measurement or observation of the turbidity of the reaction mixture without gel electrophoresis. IC/RT-LAMP was 100 times more sensitive than IC/RT-PCR.

The genomic DNA molecule of tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted geminivirus, was amplified from total DNA extracts of TYLCV-infected tomato (Lycopersicon esculentum) by the use of loop-mediated isothermal amplification (LAMP). The procedure was also used to amplify TYLCV DNA from total DNA extracts of individual whiteflies (Bemisia tabaci) that had fed on TYLCV-infected plants. One of the characteristics of the LAMP method is its ability to synthesize an extremely large amount of DNA. Accordingly, a large amount of by-product, pyrophosphate ion, is produced yielding a white precipitate of magnesium pyrophosphate in the reaction mixture. The presence or absence of this white precipitate allows easy detection of amplification of TYLCV genomic DNA without gel electrophoresis.

We describe a technique for repair of the patellar tendon rupture after total knee arthroplasty (TKA), using the Leeds-Keio (L-K) prosthetic ligament. Two patients were followed for a mean of 38 months. Knee extension power and improved function were obtained in both cases. Use of the L-K ligament is a treatment option for repair of patellar tendon rupture following TKA.

One hundred fifteen asymptomatic Japanese volunteers aged from 13 to 76 years were examined by magnetic resonance imaging (MRI) to evaluate age-related meniscal degeneration and to determine the prevalence of discoid menisci. Each meniscus was graded in the anterior and posterior portions according to intrameniscal MRI signals. Discoid meniscus was diagnosed if a bow-tie configuration was noted on three or more contiguous sagittal sections. Meniscal abnormalities on MRI became more prevalent with age in both men and women. The posterior horn of the medial meniscus showed a significantly higher prevalence of degeneration than other parts of the meniscus. Discoid menisci were noted in 15 subjects (13% prevalence), always representing the lateral meniscus. Subchondral changes were observed in 13 subjects more than 40 years old, mostly women, and were located in the medial compartment. These abnormalities were not correlated with severity of degeneration in the posterior portion of the medial meniscus. This study demonstrates considerable prevalence of meniscal abnormalities in asymptomatic Japanese subjects.

Achilles tendinous collagen fibrils insert into the calcaneus by first passing through a zone that is defined histologically as fibrocartilaginous. This zone consists of four regions: tendon proper, non-mineralized and mineralized fibrocartilage and bone. The function of this zone has not yet been clearly defined. To gain more insight into the role of this fibrocartilaginous zone, collagens present in the zone of the Achilles tendon-calcaneus interface were isolated and characterized. Types II, IX and X collagens were identified in the pepsin digests of the tissue harvested from the bovine Achilles tendon-calcaneus interface. Western blotting using specific antisera to types II, IX and X collagens confirmed the identity of these collagens. Immunofluorescence localization placed type X collagen predominantly in the mineralized zone of the tendon-calcaneus junction, while type IX collagen was distributed throughout the the insertion site. The presence of the cartilage-specific collagens at the Achilles tendon-calcaneus-interface suggests that this zone is cartilaginous in nature. The presence of type X collagen at this junction is not clear, but our present findings go along with the previous report which showed that type X collagen is present in the mineralized zone of the medial collateral ligament femoral insertion site. These data suggest that type X collagen may be a resident of mineralized fibrocartilaginous zones of tendon or ligament-bone junctions and may participate in anchoring ligament or tendon to bone.

Experimental middle ear barotrauma was studied morphologically. White guinea pigs were placed in an experimental hyperbaric chamber, and middle ear barotrauma was created by increasing the pressure in the hyperbaric chamber from 1 atmosphere absolute (ATA) to 2 ATA using pure oxygen, maintaining the pressure at 2 ATA for 10 minutes, then again reducing the pressure to 1 ATA. Selected experimental animals were decapitated immediately after, one day after, one week after, or weeks after pressure loading, and their middle ears were examined by a light microscope (LM), a scanning electron microscope (SEM) and a transmission electron microscope (TEM). Hemorrhaging in the tympanic cavity immediately after pressure exposure was apparent even macroscopically. LM also revealed evidence of submucous hemorrhage. Submucous edema was seen in the "one week after" cases. SEM showed a minor loss of cilia in some ciliated cells just after the experiment. In nonciliated cells, the terminal web was somewhat indistinct in the "one week after" cases. TEM also indicated a minor loss of cilia in some ciliated cells in "one day after" cases as well as apparent vacuoles within the cells. These findings suggest that although trauma during compression is more marked than during decompression, recovery from this damage progressed with time.

A 46-year-old man was admitted to our clinic because of acute heart failure. Six years before admission he was pointed out cardiomegary and hematuria. One year later, he was diagnosed as having jugular foramen syndrome. On admission, he had a fever and dyspnea. Pansystolic blowing murmur was audible at the apex. The chest ratio on his chest X-ray was 52.5%. An electrocardiogram showed left ventricular hypertrophy. An echocardiogram showed marked dilatation and severe dysfunction of left ventricle. Radionuclide scanning with technetium 99 m pyrophosphate identified inflammatory change in the apex. Myocardial biopsy showed fibrotic degeneration and IgG deposits in myocardium. Blood examination showed anemia, lymphopenia. positive anti-nuclear antibody (1000 times, shaggy pattern), positive anti ds-DNA antibody and hypocomplementemia. Furthermore, proteinuria was pointed out. Renal biopsy showed focal segmental glomerulonephritis with active necrotizing lesion (type III nephritis). Lupus myocarditis and nephritis was diagnosed. After prednisolone (80 mg/day) was administered. left ventricular function and hypocomplementemia improved. The ACE inhibitor was also used for proteinuria. In spite of a little amount of blood transfusion, he showed hepatic hemosiderosis. We suspect that the cause of hemosiderosis was related chronic inflammation of active lupus. It was treated with Erythropoietin.

Bleeding related to systemic heparinization has been considered one of the major complications associated with extracorporeal membrane oxygenation (ECMO). Development of the heparin-bonded system will be essential in reducing hemorrhagic complications, but has not yet been clinically proven. The authors chose an alternative approach of making a difference in the activated clotting time (ACT) values between the patient and the ECMO circuit, and decreased only the patient's ACT value, while keeping the value of the ECMO circuit at an ideal level. For this purpose, we have used a very short-life anticoagulant, Nafamostat Mesilate (FUT), while decreasing the dose of heparin. FUT is a synthetic protease inhibitor that has been found to inhibit various kinds of enzyme activities for coagulation. Twelve newborns who had some hemorrhagic complications at various sites before or during ECMO, were selected to receive FUT. The heparin dose was decreased after FUT administration into the drainage route. FUT and heparin doses were regulated to maintain the ACT value at the reinfusion route at 190 to 220 seconds. ACT values at the drainage and the reinfusion routes were simultaneously measured. The average time on FUT was 100.3 +/- 86.3 (SD) hours. The average dose of FUT was 0.48 +/- 0.22 mg/kg/h, and that of heparin was 21.0 +/- 7.5 U/kg/h. The average ACT value at the reinfusion route was 205.7 +/- 14.0 seconds compared with that at the drainage route of 178.5 +/- 11.8. The difference was statistically significant (P < .001). The average difference in ACT values between both routes was 27.1 +/- 7.9 seconds. The bleeding was well controlled by FUT administration in 8 of 12 cases. This report may represent the first clinical use of FUT in neonatal ECMO, and serve as a preliminary study.

Background: Autoimmune mechanisms have been implicated in the pathogenesis of chronic ongoing myocarditis. To investigate this relation, we used an A/J mouse model inoculated with coxsackievirus B3 and determined whether myocarditis would be transferred to normal hearts that were heterotopically transplanted., Methods and Results: Inbred 3-week-old A/J mice were inoculated intraperitoneally with coxsackievirus B3 (Nancy strain; 2 x 10(4) plaque-forming units) and housed for > 60 days. The presence of the viral genome in the myocardium was determined by the polymerase chain reaction with primers specific for the 5' end of the coxsackievirus B3 genome performed at 40, 50, or 60 days after inoculation. Normal A/J mouse hearts were transplanted into the same strain of mice without myocarditis (group A) and into mice with chronic ongoing myocarditis (group B). The hearts were evaluated histologically 2 weeks after transplantation. Conventional histological examination of infiltrated T cells and macrophages was performed, and the expression of intercellular adhesion molecule-1, major histocompatibility complex (MHC) class I antigen, and MHC class II antigen was evaluated by immunoenzymatic staining. The concentrations of interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor (TNF-alpha) in the grafts were measured with an ELISA. The viral RNA genomes were not detected in the mice with chronic ongoing myocarditis, but their transplanted hearts did show myocarditis. In the hearts with induced myocarditis, infiltrated mononuclear cells consisted of CD4+ T cells, CD8+ T cells (CD4+ cell number > CD8+ cell number), and macrophages. Intercellular adhesion molecule-1, MHC class I antigen, and MHC class II antigen were expressed in the vascular endothelial cells and myocardial cells in and around the infiltrated lesions. The concentrations of IL-1 alpha and TNF-alpha in group B were significantly higher than those in group A (group A versus group B: IL-1 alpha, 125 +/- 35 versus 180 +/- 34 pg/mL; TNF-alpha, 45 +/- 15 versus 96 +/- 40 pg/mL; P < .05)., Conclusions: Results suggest that an autoimmune response may play a key role in the progression of chronic ongoing myocarditis.

To clarify the pathogenetic mechanism of chronic ongoing myocarditis, we produced Coxsackievirus B3-induced myocarditis in A/J mice and immunopathologically examined the microcirculation in the chronic phase of myocarditis. Forty-two 3-week-old A/J mice were inoculated intraperitoneally with Coxsackievirus B3 (Nancy strain) 2 x 10(4) PFU (plaque-forming units) and sacrificed 7, 14, 21, 50, 90, or 120 days later. To evaluate myocardial microcirculation, 18 of the hearts were perfused from the thoracic aorta with warm 2% gelatin/carbon solution. The remaining hearts were quickly frozen for immunologic analysis with an enzyme immunostaining assay using monoclonal antibodies against CD4, CD8, macrophages, intercellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex class I or II. The presence of viral RNA genome in the myocardium at 40, 50, or 60 days after inoculation was evaluated using the polymerase chain reaction. The lesions in chronic ongoing myocarditis consisted of myocardial damage, myocardial calcification, interstitial fibrosis, and infiltration of mononuclear cells. These infiltrated lymphocytes were predominantly CD4+ T cells. Furthermore, microvascular abnormalities, including dilatation, tortuosity, constriction, and abrupt termination, were observed around the lesions. There was marked infiltration by mononuclear cells around the microvessels. ICAM-1 was strongly expressed in the endothelial cells of the vessels. Coxsackie B3 viral genome was not detected in the myocardium of mice with chronic ongoing myocarditis in each stage examined. These results suggest that an autoimmune mechanism is involved in the persistent inflammation seen in chronic ongoing myocarditis.

In order to investigate the mechanism of inner ear barotrauma, guinea pigs, with bilateral eustachian tube occlusion, were subjected to decompression and compression between 760 and 460 mmHg in a hypobaric pressure chamber. We divided the guinea pigs into two groups, A and B. Group A showed normal eustachian tubes, and group B showed bilaterally occluded eustachian tubes. Group B animals were divided into three types according to the rates of compression and decompression. After pressure loading, morphological changes in the hair cells of the organ of Corti were studied by means of scanning electron microscopy. There was no damage to hair cells in the setting of normal eustachian tube function, as in group A. On the other hand, mild to severe hair cell damage was observed with rapid decompression in group B. This observation suggests that relative positive pressure in the middle ear cavity is an important factor in inner ear barotrauma. The mechanism of hair cell damage due to inner ear barotrauma is presumed to be distortion of the organ of Corti caused by a difference in pressure between perilymph and endolymph resulting in injury to the stereocilica.

Morphological vestibular changes caused by barotrauma were studied in guinea pigs. Animals were exposed to rapid decompression from 2 absolute atmospheric pressures (ATA) to 1 ATA, which causes inner ear barotrauma in the guinea pig. During decompression, spontaneous nystagmus was recorded, which consisted of irritative symptoms initially, followed by paralytic nystagmus. After pressure loading and observation to confirm the absence of Preyer's reflex with vertigo, the animals were tested for caloric nystagmus using ice water and then sacrificed at varying intervals. Then, morphological changes in vestibular organs and the organ of Corti were studied. Half of the experimental animals showed canal paresis on caloric testing. Damage to the organ of Corti was severe while that to vestibular organs was very slight. Damage to the sensory cells of the vestibular organs was not clear on light microscopy, despite a partial collapse of labyrinthine membranes. Under scanning electron microscopy, local damage was observed in a portion of the crista ampullaris of the semicircular canals. In this area, incomplete or complete disappearance of kinocilia and stereocilia, similar to that seen after rotatostimulation, was observed. However, no damage to sensory hairs was seen in the utricles and saccules. The observed vestibular organ damage, resulting from inner ear barotrauma, suggested effects on endolymphatic flow.

The question of whether the etiology of DCM is immune or autoimmune has been increasingly discussed. Abnormal findings on humoral immunity in DCM were investigated, especially those regarding anti-heart antibodies (AHA), IgG subclasses and soluble interleukin-2 receptor (sIL-2R). The heterophile type AHA was detected in 64.7% of cases by the indirect immunofluorescence technique (IF) with rat heart, by indirect IF with human heart AHA in 57.8% of cases, and by thin-layer chromatogram with human glycolipids AHA in 44% of cases. Also, 57.1% of the specimens were found to bind IgG on perimyocytes by direct IF with biopsy specimens taken from patients with DCM. The epitope of an antigen which reacted with the heterophile type AHA is a Gal alpha 1-3Gal structure. 200 Kd, 70 Kd and 40 Kd antigens were reacted with AHA detected by indirect IF with human heart. The possible mechanisms of AHA in the pathogenesis could be either complement dependent cytotoxicity or interference to cardiac metabolism. The concentration of sIL-2R and IgG3 in sera from patients with DCM were elevated. These results suggest that immunological abnormalities occur continuously in DCM.

To identify genetic factors in the immune system which control the susceptibility to dilated cardiomyopathy (DCM), HLA class II DNA typing was performed in 61 Japanese patients, using PCR/SSO probe analyses. The frequencies of HLA-DQB1*0503 (15% vs 5%; RR = 3.06, chi 2 = 7.19) and DQB1*0604 (21% vs 10%; RR = 2.41, chi 2 = 6.20) were significantly increased and that of HLA-DQB1*0502 (RR = 1.74) was slightly increased in the DCM patients. The frequency of DQB1*0303 (16% vs 31%; RR = 0.44, chi 2 = 5.16) was significantly decreased in the patients. The increased HLA-DQB1 alleles have a histidine residue in common at the 30th codon for the HLA-DQ beta chain. Among the genetic markers studied by Southern blot analyses, IGLV (immunoglobulin lambda light chain, pV3.3) showed a strong association with DCM, i.e. A2/A2 genotype was found in 37.7% of patients whereas it was observed in only 18.9% of the control subjects (RR = 2.6, chi 2 = 7.77). The frequency of this genotype was higher in patients under age 45 years at the time of diagnosis (45.5%, RR = 3.6, chi 2 = 10.02). These results suggest that HLA and immunoglobulin genes are closely linked to susceptibility to DCM.

Recently, ceramic prostheses for ossicular replacement have become very popular in middle ear surgery. These prostheses have a good affinity with the surrounding tissue. But, it is difficult to identify the position of these prostheses postoperatively. We used magnified tomography in order to identify the implanted ceramic prostheses, and to determine whether these prostheses were in the appropriate position. In the case of patients without stapes, we usually use Apaceram type T, which restores ossicular continuity to the inner ear. When the shaft of the ceramic prosthesis is placed on the oval window niche, the overhanging Fallopian canal obstructs the tip of the prosthetic shaft to obtain the appropriate place in the oval window. The angle of the shaft should be slightly tilted inferiorly against the medial wall of the middle ear cavity.

A 25 year old man was admitted to our hospital on June 1, 1989 because of general fatigue and pleural effusion. He had noticed purpuras and nodules on his bilateral lower legs in July 1988. He was admitted to Nihon University Hospital and was diagnosed as allergic granulomatous angiitis. Methylprednisolone was administered. In March, 1989, a nodular shadow was detected in his lung CT films. From the findings of TBLB a granulomatous lesion was suspected. However, his clinical course was rather stable so he was discharged from the hospital. Two months later, he was suffered from fever and general fatigue. On his chest X ray film pleural effusion was detected in the lower part of his left lung. The nodular shadow was increasing gradually. Laboratory findings on his admission showed leukocytosis, thrombocytosis, elevated CRP and a high titer of RA factor. He was diagnosed as hypersensitivity angiitis in a broad sense based on his biopsy findings of the skin lesion. Furthermore, the diagnosis of an early stage of Wegener's granulomatosis (WG) or limited type of WG was also made from the clinical course of his lung lesion. The combination therapy with cyclophosphamide and methylprednisolone was started. It was so effective that the nodule of his left lung was almost disappeared. It seems that this case is one of the polyangiitis overlap syndrome proposed by Fauci.

To study the effects of a persistent perilymph fistula on the cochlea, a small cannula was inserted into the scala tympani of the basal turn of cochlea in guinea pigs. A month later, cochlear morphology and blood flow were studied using either histological evaluation or the microsphere surface preparation technique. Some animals showed no cochlear morphologic changes or no cochlear blood-flow reduction, even if tubal patency was maintained and perilymph leakage lasted for 1 month. This suggests that a prolonged perilymph fistula, per se, causes no permanent cochlear damage. However, in some animals, hair cell damage and cochlear blood-flow disorders were observed. These observations and the causes of hearing loss in clinical cases of perilymph fistula were studied.

In order to investigate inner ear barotrauma, guinea pigs were subjected to rapid decompression between 2 absolute pressure (ATA) and 1 ATA in a chamber. After pressure loading and observation for absence of Preyer's reflex, they were sacrificed immediately, 1 day, 1 week and 1 month later, respectively. Then, morphological changes of the organ of Corti and stria vascularis were studied under TEM and SEM. The immediate features noted were fracture of stereocilia with minimal intracellular changes. One day later, there was marked degeneration of outer hair cells and expansion of supporting cells. The damage to stereocilia clearly preceded morphological alterations within hair cell bodies and cannot be interpreted as arising secondary to hair cell degeneration. Most of outer hair cells eventually disappeared and were replaced by supporting cells. Inner hair cells degenerated slowly; 1 month later, some of them remained almost intact, despite disappearance of stereocilia. The continuity of reticular lamina was maintained not only immediately but also through the period of hair cell degeneration, thus preventing any leakage of endolymph into the organ of Corti. There was reversible dendritic swelling of inner hair cells immediately following the trauma. No changes of stria vascularis were observed over passage of time. The mechanism of hair cell damage due to inner ear barotrauma is presumed to be a deformity of the organ of Corti caused by pressure discrepancy between perilymph and endolymph resulting in an injury to stereocilia.

Relationship of circulating immune complexes to some inner ear disorders has been suggested recently. Thus, bovine serum albumin (BSA) nephritis, a model of chronic serum sickness, was made and morphological, immunological studies of cochlea were performed using light microscopy, transmission electron microscopy and immunofluorescent antibody methods. Group 1 consisted of animal sensitized with daily BSA injections for 90 days; in group 2, BSA was administered for 70 days, daily, and after a three-weeks interval, injections were continued for another 21 days; group 3 consisted of animals sensitized with daily BSA injections for 100 days followed by BSA injection into the facial nerve via the stylo-mastoid foramen. The animals (group 3) were sacrificed on day 1 and 4 following facial nerve sensitization for examination of cochlear morphological changes. No morphological changes were observed (group 1). Protrusion of marginal cells into endolymphatic space was seen, because of swelling of endoplasmic reticulum of the cells of stria vascularis's (group 2). Slight hydrops was present mainly in the basal turn (group 3). No IgG deposits were found in all groups. It appears that the changes in group 2 were caused by rather chronic nephritis than immune reaction. We conclude that damage induced in the cochlea is negligible compared to the extensive lesions observed in the kidney in chronic serum sickness.

Cannula occlusion is a life-threatening postoperative complication of tracheostomy. Current management largely relies on nursing care for prevention of fatalities because no proven mechanical, machine-based support monitoring exists. The objective of this paper was to address the problem of monitoring the state of cannula patency, based on analysis of airflow acoustic spectral patterns in tracheostomized adult patients in the patent and partially occluded cannula. Tracheal airflow sounds were picked up via a condenser microphone air-coupled to the skin just below the tracheal stoma. Signal output from Mic was amplified, high-pass filtered, digital tape-recorded, and analyzed on a mainframe computer. Although airflow frequencies for patient cannulae were predominantly low-pitched (0.1 to 0.3 kHz), occluded tubes had discrete high-pitched spectral peaks (1.3 to 1.6 kHz). These results suggest that frequency analysis of airflow sounds can identify a change in the status of cannula patency.

A case of cardiac sarcoidosis with refractory ventricular tachycardia is presented. This case was followed up for 6 years and was autopsied. The cephalad portion of the interventricular septum and the right ventricular outflow tract were markedly thinned, and sarcoid granulomas were also found in the lung.

The production of myocardial lesions and circulating anti-heart antibodies has been studied in rats injected with homologous heart extracts emulsified with the Freund's complete adjuvant and the DTP vaccine. Six of 7 rats showed lesions of focal myocarditis, while only in one of them circulating anti-heart antibodies were detected. There was no correlation between the presence of circulating anti-heart antibodies and that of myocardial lesions. These findings suggested that the circulating anti-heart antibodies may play little or no role in the pathogenesis of this experimental myocarditis.

In our previous study, a high rate of "heterophile antibody" was found in idiopathic cardiomyopathy. In the present investigation the cytotoxic activity of this antibody was examined in cultured myocardial cells. Trypsin treated cells cultured for 3 days were used in the two-step method of the complement dependent cytotoxicity test. The trypan blue dye-exclusion method was utilized to determine the percentage of surviving cells, and the cytotoxic effect was expressed by the cytotoxic index (CI). With fresh rabbit serum as complement, the mean CI value of the heterophile antibody positive sera was 20.7, which was in contrast to the value of anti-heart antibodies negative sera (p less than 0.01). As the cytotoxic effect to the heterophile antibody was absent without the complement, it may be complement dependent. Thus, our results suggest that the heterophile antibody may play a role in the pathogenesis or be an incremental factor of idiopathic cardiomyopathy.

Circulating anti-heart antibodies (C-AHAb) were examined in the sera of 132 patients with heart diseases and of 65 normal healthy controls using an indirect immunofluorescent technique. The patient groups consisted of 31 with dilated cardiomyopathy, 22 with hypertrophic cardiomyopathy, 38 with myocardial infarction, 15 with angina pectoris and 26 with rheumatic valvular disease. The indirect immunofluorescent staining patterns were classified into 4 types: "heterophile", "intracellular", "intercalated disc" and "nuclear" patterns. The positive incidence of C-AHAb was 81% in dilated cardiomyopathy, 77% in hypertrophic cardiomyopathy, 65% in rheumatic valvular disease, 58% in myocardial infarction, 40% in angina pectoris and 39% in normal healthy controls. A high rate of "heterophile" antibodies was found in the idiopathic cardiomyopathy group: 65% in dilated cardiomyopathy and 55% in hypertrophic cardiomyopathy. The intracellular staining pattern was present in 42% of rheumatic valvular disease. These findings suggest that positive C-AHAb and staining patterns may be useful in the assessment of the pathogenesis of certain types of heart diseases.

To determine the immune disturbances involved in the pathogenesis of dilated cardiomyopathy (DCM), the conditions in humoral and cellular immunity of DCM patients were studied. To examine the status in humoral immunity in DCM patients, circulating anti-heart antibodies (AHAbs) were analyzed. Circulating-AHAbs were classified into 4 types "heterophile-like", "intracellular", "intercalated disc" and "nuclear" patterns by indirect immunofluorescence technique, and a high rate of heterophile-like antibody was found in DCM. The cytotoxicity of this antibody was examined in cultured myocardial cells using the two-step method of the complement dependent cytotoxic test. The mean cytotoxic index (CI) value of the heterophile-like antibody positive sera was 22.3, which showed an apparent cytotoxicity against the cultured cells and it may be complement dependent. In addition, the frequency of lymphocyte subsets using monoclonal antibodies (OKT4, OKT8, Leu7, Leu11) and natural killer (NK) activity were evaluated to determine whether DCM patients had an imbalance in cellular immune reactions, which support the hypothesis of an immune disturbance as the pathological mechanism of DCM. The peripheral lymphocyte counts were significantly lower in patients with DCM (1737 +/- 874/mm3) than in normal controls (NC, 2088 +/- 556/mm3) and in patients with ischemic heart diseases (IHD, 2395 +/- 469/mm3, both p less than 0.01). The percentage of T, B, OKT4 and OKT8 positive cells was not statistically different among DCM, IHD, and NC groups, whereas the percentage of T gamma cells was significantly reduced in DCM patients (6.5 +/- 5.0%, p less than 0.05). NK functional activity as tested in DCM patients was frequently deficient (24.1 +/- 16.7% in DCM, 36.7 +/- 12.2% in NC). After 4-hr incubation with recombinant interleukin2 (rIL2), rIL2 induced the enhancement of NK activity in 3 out of 4 DCM patients with low NK activity, although, there was a non-responder to rIL2 in this group. These results suggested that DCM patients have a low IL2 production and/or less numbers of mature cells with NK cell function. Therefore, an imbalance in humoral and cellular immune reactions may cause insidious myocardial damage and subsequently lead to development of DCM.

In patients with chronic obstructive pulmonary disease, quantitative analysis of serum IgG, IgA and IgM were carried out by use of the radial immunodiffusion method. The concentration of IgE in sera was also determined by the radioactive radial immunodiffusion method. The mean value of serum IgG level in the group of cor pulmonale was higher than that in groups of emphysema, asthma and normal subjects. The mean value of serum IgA level was higher in two groups of cor pulmonale and emphysema than in the group of asthma and of normal subjects. There was no difference of serum IgM levels between these four groups. The mean value of serum IgE in the group of allergic asthma was higher than that in the group of non-allergic asthma or normal subjects and an elevated serum IgE level was also found in patients with cor pulmonale. Findings of this study suggested that the raised level of serum IgG in patients with chronic cor pulmonale might reflect production of antiheart antibodies against cardiac tissue.

The present study has indicated the presence of 3 heart specific antigens, using homologous and heterologous antibodies produced in immunized rabbits. Two of these antigenic proteins exhibit restricted organ specificity for heart and the other one shares by heart and kidney. One of these 2 heart specific antigens reacts to both homologous and heterologous anti-heart sera absorbed with kidney, while the other reacts to only heterologous anti-heart sera. The former has an electrophoretic mobility corresponding to that of serum beta-globulin, and is found to have a molecular weight of about 175,000. The latter has the same electrophoretic mobility as that of alpha2-globulin and is found to have a molecular weight of about 50,000.

The production of myocardial lesions and circulating anti-heart antibodies has been studied in the present experiment. Twenty-one of Wistar rats were divided into 3 groups: 9 rats immunized with heterologous heart extracts, Freund's complete adjuvant and diphtheria-tetanus-pertussis (DTP) vaccine, 8 rats injected phosphate buffered saline adjuvant mixture with DTP vaccine and 4 rats untreated as control. Injection of heterologous heart extracts emulsified with Freund's complete adjuvant and DTP vaccine developed focal myocardial lesions in all of the subjected rats. The lesions were characterized by focal and diffuse interstitial cellular infiltration with myocardial necrosis. The circulating antibodies reacted to rat heart extracts were detected in 77.8% of the subjects. The antigen, reacted with the antibodies, located intracellular spaces in myocardium. The severity of cardiac lesions were not functional to the titer of circulating anti-heart antibodies may play little role in the pathogenesis of myocardial damage at least in this experimental myocarditis.

An acquired defect or damage of a subpopulation of suppressor T lymphocytes is reported in connection with autoimmune diseases. In the present study, the role of immunity was examined in 7 patients with dilated cardiomyopathy (DCM). The frequency of lymphocyte subsets using monoclonal antibodies and natural killer (NK) cell activity was evaluated to determine whether DCM patients had lymphocyte abnormalities that would support the hypothesis that the pathological mechanism of DCM is an immune disturbance. The peripheral lymphocyte counts were significantly lower in patients with DCM and higher in patients with ischemic heart disease (IHD) than in normal controls (NC) (p less than 0.01). The percentage of T cells, B cells, OKT4 and OKT8 positive cells was not statistically different among the three groups studied here, whereas the percentage of T gamma cells was significantly reduced in DCM patients (p less than 0.05). NK cell functional activity as tested in DCM and IHD patients was frequently deficient (22.1 +/- 19.3% in DCM, 13.8 +/- 3.0% in IHD, 37.4 +/- 12.7% in NC). Our results suggest that an imbalance in cellular immune reactions partly explain the pathogenesis of DCM.

To determine whether bunazosin (alpha 1-adrenoceptor blocking agent) can alter the hemodynamic profile of chronic heart failure secondary to myocardial infarction, the drug was administered to Wistar rats 4 weeks after coronary ligation, and continued for 4 weeks. In rats without bunazosin treatment, the left ventricular end-diastolic pressure (LVEDP) and the total vascular resistance index (TVRI) increased as a function of infarct size, while the cardiac index (CI) decreased. But in infarcted rats with bunazosin treatment, the mean aortic pressure and TVRI were reduced, the LVEDP was modestly lessened, and the CI was maintained. The greatest increase in CI after the treatment occurred in rats with infarcts of small and moderate size. Thus, long-term therapy with bunazosin improved LV dysfunction, relative to the size of infarction. This study suggests the beneficial effects of bunazosin therapy in patients with chronic heart failure.

Mouse monoclonal antibody CC3C195, which detects elevated levels of its antigen in sera from many patients with colon and pancreatic cancer, binds with high affinity to the sialylated human Lea blood group antigen NeuAc alpha 2-3Gal beta 1-3 [Fuc alpha 1-4]GlcNac . . . and with lower affinity to the Lea blood group antigen itself.