Multifocal granulomatous hepatitis caused by Actinobacillus pleuropneumoniae serotype 2 in slaughter pigs.

In a survey of 66 894 slaughter pigs, 11 animals from three farms were found to have multifocal granulomatous lesions in the liver, caused by Actinobacillus pleuropneumoniae serotype 2. The lesions consisted of epithelioid cells and multinucleated giant cells, with asteroid bodies and discernible gram-negative bacteria. Lymph nodes and spleen were occasionally affected. The results suggested that haematogenous spread had occurred from pre-existing pulmonary infections.

Probe of bending motion following the 1s(-1)pi* excitation of N2O.

The doubly degenerate core-excited Pi state of N2O splits into two due to the static Renner-Teller effect. The lower state, A1, has a bent stable geometry and the molecule excited to this state starts to deform itself toward this bent geometry. To probe the effect of the potential energy surfaces of the core-excited A1 states on the nuclear motion, we measure the momenta of the three atomic ions in coincidence by means of the ion momentum imaging technique. We find that the potential energy surface affects the molecular deformation significantly. N2O in the terminal N 1s(-1)3piA1 excited state is observed to be bent more than that in the central N 1s(-1)3piA1 excited state. This means that N2O in the terminal N 1s(-1)3piA1 excited state bends faster than that in the central N 1s(-1)3piA1 excited state. When the excitation energy is decreased within the 1s(-1)3pi resonances, the nuclear motion in the A1 states becomes faster. This is interpreted by the notion that the excitation occurs onto the steeper slope part of the potential energy surface of the excited state for the lower excitation energy. The branching ratio of the A1 excitation increases with the decrease in the excitation energy.

Effects of trematode infection on metabolism and activity in a freshwater snail, Semisulcospira libertina.

Changes in the metabolism and activity of the freshwater snail Semisulcospira libertina infected with larval trematodes were studied experimentally. In snails up to 11 mm in shell width, crawling distance, feeding frequency, and the proportion of individuals located on vertical walls did not differ among snails infected with mature or immature cercariae, or uninfected snails (p > 0.05). In snails larger than 11 mm, individuals infected with mature cercariae tended to feed more frequently during the light period (p = 0.0081), but the distance they crawled and the proportion of individuals located on vertical walls did not differ, regardless of infection (p > 0.05). Infection with mature cercariae significantly increased the oxygen consumption rate (p = 0.016), which was measured only in the large size.

Expression and function of galectin-3, a beta-galactoside-binding protein in activated T lymphocytes.

A soluble beta-galactoside-binding lectin, galectin-3 has been shown to be involved in cell adhesion and activation of immune cells. Although galectin-3 is known to be expressed in various types of cells, it has not been shown whether galectin-3 is expressed in T lymphocytes. We present evidence here that galectin-3 is expressed in activated murine T lymphocytes including CD4+ and CD8+ T cells but not in resting T cells. Galectin-3 expression was induced by anti-CD3 mAb or mitogen and enhanced by common gamma-chain signaling cytokines, IL-2, IL-4, and IL-7, in activated T lymphocytes, whereas the inflammatory cytokines including TNF-alpha and IFN-gamma did not. Galectin-3 expression and proliferation were down-regulated by withdrawal of IL-2 and gamma irradiation. Antisense but not sense phosphorothioated oligonucleotides for galectin-3 inhibited galectin-3 expression and blocked proliferation of T cells significantly. This study suggests that up-regulation of galectin-3 plays an important role in proliferation of activated T lymphocytes.

Mechanical strength of intramedullary pinning and transfragmental Kirschner wire fixation for Colles' fractures.

Four methods of distal radius fracture fixation were tested in this experimental biomechanical study of an unstable Colles' fracture model. Sixty artificial radial bones and seven pairs of cadaveric radii were used. Seven additional pairs of paired cadaveric radial epiphyses were used in separate perforation and cut-out tests. Tests with the artificial bones showed that the rigidity of the four tested fixation methods was comparable, except for proximal intramedullary pinning. The rigidity of the NODE fixation in the cadaveric radii was greater than that of transfragmental pinning. The load at failure was significantly greater for the NODE system than for the Kirschner wire models. Rigidity and failure loads showed a positive correlation with bone mineral density. The current study indicates the NODE system was the strongest of the fixation methods.

Role of standardized patients in medical education.

Standardized patients are volunteers from the community who participate actively in the teaching and evaluation of our future physicians. The SP program augments medical student learning in both clinical and basic sciences by providing opportunities to practice on patients who are available and willing to be examined. SPs allow "teachable moments" to be created, rather than waited for. SP-based examinations are the most reliable and standardized methods for performance-based assessment of bedside clinical skills. The problem-based-learning (PBL) approach to medical education at JABSOM is designed to foster self-directed learning, interpersonal skills and early hands-on clinical experiences. JABSOM SP experiences are integrated within the PBL curriculum. The CCS and SPs are now vital components of the JABSOM educational model.

Hemodialysis-related subacromial lesion: diagnostic imaging and minimally invasive treatment.

BACKGROUND: In an attempt to refine the diagnosis and surgical treatment system for spontaneous shoulder pain in supine position (SPSP) in long-term hemodialysis (LTHD) patients we reviewed shoulders of patients during a 4-year period. PATIENTS AND METHODS: Clinical findings, imaging findings and operative results (average duration of follow-up was 39 months) on both shoulders of 110 patients (Shigei Hospital, Okayama, Japan) were analyzed to identify the cause of SPSP in order to accurately differentiate it from other shoulder pains, and to select the proper treatment method. RESULTS: As the hemodialysis period lengthened, increase in the incidence and severity of bone and soft tissue changes (radiolucency, joint destruction, increase of thickness of subacromial bursa and rotator cuff, and synovial proliferation) in the shoulder induced by dialysis-related amyloidosis were noted in roentgenography, ultrasonography and magnetic resonance imaging. SPSP occurred in the LTHD patients. An increase in subacromial bursa and rotator cuff thickness correlated with SPSP. A decrease in subacromial space correlated with change in position and SPSP. An increase of subacromial pressure was judged to be the cause of SPSP, and subacromial decompression by coraco-acromial ligament release was effective for relieving SPSP. When massive synovial proliferation or invasion of granulation tissues was observed in the glenohumeral joint, arthroscopic debridement was necessary. CONCLUSION: SPSP is a sensitive indicator of a distinct subacromial lesion frequently found in LTHD patients. These lesions can be treated by minimally invasive endoscopic coraco-acromial ligament release, resulting in marked pain relief.

Local secretion of IFN-gamma induces an antitumor response: comparison between T cells plus IL-2 and IFN-gamma transfected tumor cells.

Previously we described that the adoptive transfer of tumor-infiltrating lymphocytes (TIL) + interleukin-2 (IL-2) leads to eradication of established methylcholanthrene (MCA)-105 fibrosarcoma pulmonary metastases in a congenic murine model. The in vivo efficacy of TIL was associated with their ability to secrete interferon-gamma (IFN-gamma), and to a lesser extent granulocyte-macrophage colony-stimulating factor. The local secretion of these cytokines resulted in recruitment of naive host immune cells to the tumor and eventually in a successful host antitumor immune response. In the present study, to further evaluate the role of IFN-gamma in the induction of a host antitumor immune response, we compared the treatment efficacy of adoptively transferred T cells and IFN-gamma gene transfected tumor cells (MCA-105/IFN-gamma) as delivery systems of IFN-gamma. Treatment with TIL-IL-2 or irradiated MCA-105/IFN-gamma induced a similar reduction in pulmonary metastases of MCA-105 tumor. In contrast, irradiated wild-type MCA-105 or TIL from IFN-gamma gene knockout mice did not cause tumor eradication. MCA-105 tumor-bearing mice treated with MCA-205/IFN-gamma showed a partial reduction in the number of pulmonary metastases. Histologically, lungs of successfully treated mice showed that initially activated macrophages expressing inducible nitric oxide synthase (iNOS) and dendritic cells infiltrated the tumor bed. Subsequently, CD4+ and CD8+ T cells infiltrated tumors. The therapeutic efficacy of IFN-gamma transfected tumor cells was eliminated when either CD4+ T cells or CD8+ T cells were depleted. These results suggest that local secretion of IFN-gamma induces a tumor-specific host antitumor immune response mediated through activated macrophages, dendritic cells, and tumor-specific T cells. This may be a common component of successful immunotherapy.

Ultrasonographic analysis of shoulder rotator cuff tears.

Seventy-nine shoulders suspected of rotator cuff tears were examined by ultrasonography (US) and forty-three received surgery. Long and short axis scans were performed and findings of each were separately classified according to a five-grade system, and the results were correlated with the actual extent of tear observed during surgery. Internal echogenicity and subacromial impingement were analyzed before and after surgery. A accuracy of US in detecting rotator cuff tears was analyzed. In addition, the correlation between cuff shape observed by US before surgery and actual shape observed during surgery was assessed. It was noted that cuff thinning and abnormalities in shape did not recover to normal after surgery. However, in the cases of discontinuities observed by US before surgery, US findings indicated that the torn cuff was anchored to the greater tuberosity and functional during active motion. Although post-operative US findings were not normal, clinical results were good in most cases. Sensitivity of US for detecting rotator cuff tear was 100% and specificity 94%. US is non-invasive, cost effective and allows the physician to examine the joint while it is in motion. Therefore, at this time, we use US as a screening method for detecting rotator cuff tears. Furthermore, US allows us to check for re-tears while the joint is in motion, which is essential for accurate diagnosis.

Adequacy and long-term prognosis of endoscopic carpal tunnel release.

Forty-one hands of 37 patients with idiopathic carpal tunnel syndrome treated by endoscopic carpal tunnel release (ECTR) were followed up for more than one year after surgery. Surgical results were evaluated using Kelly's criteria, the Semmes-Weinstein test, the static and moving 2-point discrimination tests, tip-pinch strength, and motor and sensory nerve conduction studies. Clinical results, according to Kelly's criteria three months after surgery, were excellent or good in 36 hands, and fair or poor in five hands. No recovery was evident at six months and 12 months after surgery in fair and poor hands. Based on these findings, we conclude that a neurolysis of the median nerve and release of constriction of the thenar muscle branch should be performed using the conventional open technique for patients with poor results three months after ECTR if the patients are dissatisfied with ECTR results.

Tumor-specific cytokine release by donor T cells induces an effective host anti-tumor response through recruitment of host naive antigen presenting cells.

We recently reported that tumor eradication induced by immunotherapy (IT) in a congenic mouse model using tumor infiltrating lymphocytes (TIL) + recombinant interleukin-2 (rIL-2) is dependent on recruitment of naive host immune cells at the tumor sites. The recruitment of host immune cells was induced mainly through a local secretion of interferon-gamma (IFN-gamma) produced by donor T cells. We now further investigated how a non-specific inflammatory response progresses to a host T-cell-mediated tumor-specific response. In cross-over experiments using MCA-105 and MCA-205 sarcoma tumors, pulmonary metastatic disease was eradicated only in mice treated with tumor-matched TIL + rIL-2. In vitro, TIL stimulated with the tumor of origin secreted relatively high levels of IFN-gamma and granulocyte-macrophage colony stimulating factor (GM-CSF) compared to TIL stimulated with mismatched tumor cells. In lungs of tumor-bearing mice treated with matched TIL + rIL-2, significant increases in the percentages of IFN-gamma, GM-CSF and tumor necrosis factor-alpha (TNF-alpha) positive cells were detected, as well as of macrophages, natural killer (NK) cells and dendritic cells. Depletion of macrophages or NK cells did not inhibit the efficacy. In contrast, depletion of dendritic cells partially inhibited the efficacy of the treatment. Combined depletion of dendritic cells and macrophages abrogated more than 80% of the efficacy. Our data suggest that successful IT may require 3 steps: (1) release of inflammatory cytokines by donor TIL after restimulation by tumor cells; (2) infiltration of host immune cells in response to local cytokine production; and (3) activation of tumor-specific host immune cells by dendritic cells and to a lesser extent by macrophages.

Successful adoptive cellular immunotherapy is dependent on induction of a host immune response triggered by cytokine (IFN-gamma and granulocyte/macrophage colony-stimulating factor) producing donor tumor-infiltrating lymphocytes.

Adoptive immunotherapy with tumor-infiltrating lymphocytes (TIL) and systemic low dose rIL-2 effectively eradicates pulmonary metastases of the murine MCA-105 sarcoma. We described earlier that host CD8+ T cells are critical for tumor eradication and that successful treatment is associated with production of high levels of IFN-gamma and granulocyte/macrophage (GM)-CSF by donor TIL in vitro. Here, we propose the mechanism through which adoptively transferred Thy-1.1+ TIL induce a host antitumor response in congenic Thy-1.2+ tumor-bearing mice. Donor Thy-1.1+ TIL were detected at the tumor site 12 h after transfer. These Thy-1.1+ cells produced IFN-gamma and GM-CSF in situ. The percentage of Thy-1.1+ TIL at the tumor site increased up to 16.4 +/- 4.9% 24 h after transfer but decreased to undetectable levels thereafter. In contrast, the percentages of host cells producing IFN-gamma and GM-CSF continued to increase at the tumor site. These increases were significantly higher in TIL + rIL-2-treated mice compared with untreated mice and rIL-2-treated mice 48 h after TIL transfer. The appearance of IFN-gamma+ and GM-CSF+ cells was followed by a large influx of host CD4+, CD8+, and Thy-1.2+ TIL and eventually by tumor eradication. This response was tumor specific since TIL obtained from MCA-205 did not induce high levels of IFN-gamma and GM-CSF and did not induce tumor eradication of MCA-105 tumor. Coinjection of Thy-1.1+ TIL and anti-IFN-gamma or anti-GM-CSF mAb significantly inhibited antitumor efficacy of the TIL + rIL-2 treatment. We conclude that successful adoptive immunotherapy in this model is mediated through cytokine production by adoptively transferred TIL that induce a host T cell-dependent antitumor response.

Bilateral adrenal neuroblastoma.

Neuroblastoma has been recognized as the most common solid tumor of infancy and childhood. The occurrence of bilateral adrenal neuroblastoma, however, is extremely rare and only a small number of cases have been previously reported. The authors herein report the clinical, histopathological and molecular biological features of two bilateral adrenal cases out of 125 neuroblastoma patients treated at Kyushu University Hospital over a 35-year period. The clinical and histopathological data of the two cases of bilateral adrenal neuroblastoma were reviewed. In Case 1, which had multiple liver metastases, a post-mortem examination revealed bilateral adrenal involvement. In Case 2, which had been detected by mass screening, CT showed masses in both adrenal glands. No big differences in size were recognized between the tumors in either of the cases. A histopathological examination revealed rosette fibrillary type neuroblastomas in both cases. The DNA of these tumor samples stored at -80 degrees C was extracted and the number of copies of the N-myc gene was determined by a Southern blot analysis. Fourteen copies of the gene were detected in Case 1, whereas neither of the tumors in Case 2 showed any amplification. The clinical outcome, histopathological findings and N-myc gene analysis of two cases might support the variety of biological features of this rare group of neuroblastoma.

Percutaneous release for trigger finger in idiopathic and hemodialysis patients.

Sixty-seven trigger fingers of 58 idiopathic and hemodialysis patients were treated by percutaneous A1-pulley release technique. Severity of triggering was classified into five grades for treatment selection and prediction of possible results. Results were excellent in 41 fingers, good in 9, fair in 7, and poor in 10, requiring additional treatment. The results of the lower grades were better, and those of the higher grades were poor. Excellent or good results appeared to depend on the proper selection of the patients according to the grading system and confirmation of triggering disappearance just after the release. There were neither infections nor neuro-vascular deficits after treatment. Compared to conventional open release, this treatment was found to be more useful from the standpoints of ease and safety of the technique, and the patients' quick return to normal life.

Human pancreatic cancer cells (MPanc-96) recognized by autologous tumor-infiltrating lymphocytes after in vitro as well as in vivo tumor expansion.

A human tumor line designated MPanc-96 has been established from a poorly differentiated primary pancreatic adenocarcinoma. MPanc-96 has a doubling time of 27 hr and grows as a confluent monolayer in various culture media. Cytogenetic analysis of in vitro-cultured tumor cells revealed a large number of clonal chromosomal aberrations, confirming their neoplastic origin. MPanc-96 grows in SCID mice when injected s.c. Xenografts established from the tumor line had a similar histology as the primary tumor. Tumor-infiltrating lymphocytes (TILs) were isolated from the primary tumor, and cytotoxic T lymphocytes (CTLs) were generated after activation on immobilized anti-CD3 monoclonal antibody (MAb) for 48 hr, expansion in low-dose IL-2 and repeated stimulation with irradiated MPanc-96 tumor cells. The generated CTLs lysed fresh autologous tumor cells as well as in vitro and in vivo expanded tumor cells from passages 9-53, suggesting that one or more tumor-associated antigens (TAAs) are stably expressed. CTLs lysed tumor cells in an HLA-class I-restricted fashion but showed no significant cytotoxicity against autologous fibroblasts, several allogeneic pancreatic cancer cell lines or K562. Our findings may be significant for the design of an animal model for studying the mechanisms of immunotherapy in human pancreatic cancer or for the identification of TAAs in pancreatic cancer.

Guyon's canal syndrome. A different clinical presentation caused by an atypical fibrous band.

An atypical case of Guyon's canal syndrome is reported. A 40-year-old woman with hypoaesthesia and claw-finger deformity of the little finger, hypothenar muscle atrophy but no first dorsal interosseous muscle atrophy, underwent release of a fibrous band to decompress the ulnar nerve at the deep branch just proximal to the branch to the abductor digiti minimi muscle. The claw-finger deformity recovered 1 week after surgery.

Improved in vivo efficacy of tumor-infiltrating lymphocytes after restimulation with irradiated tumor cells in vitro.

BACKGROUND: We investigated different culture conditions for tumor-infiltrating lymphocytes (TILs) with regard to proliferation, phenotypic changes, in vitro cytotoxicity, and in vivo therapeutic efficacy. METHODS: After enzymatic digestion of the murine fibrosarcoma, MCA-105, TIL cultures were initiated as pure lymphocyte (groups 1 and 2) or mixed lymphocyte/tumor suspensions (groups 3 and 4). Group I TILs were grown in culture medium containing 100 IU/ml recombinant interleukin-2 (rIL-2). Group 2 TILs were stimulated with solid-phase anti-CD3 monoclonal antibody (mAb) for 48 h and cultured in rIL-2 (100 IU/ml)-containing medium. Group 3, which consisted initially of a surplus of tumor cells, received the same treatment as group 2. Group 4 was also activated with anti-CD3 mAb and rIL-2 but was additionally restimulated weekly with irradiated tumor cells (TILs to tumor, 20:1). RESULTS: Groups 1 and 2 showed up to twofold higher increases in TIL numbers compared with groups 3 and 4 by the end of culture week 5. Although the original lymphocyte/tumor cell suspension consisted of 12.0 +/- 3.8% CD4+ T cells and 5.3 +/- 3.3% CD8+ T cells, all four TIL cultures showed approximately 80% CD8+ TILs and no CD4+ TILs by the end of culture week 4. In vitro cytotoxicity did not correlate with in vivo efficacy of the examined TIL cultures. By using the MCA-105 pulmonary metastases model in C57BL/6 mice, only suboptimal doses of TILs (2 x 10(6)) from group 4, which had been restimulated weekly with irradiated tumor, showed significant tumor eradication compared with all other treatment groups (p < 0.01). CONCLUSIONS: We conclude that in vitro tumor restimulation of TILs improves in vivo efficacy, most likely through the education of tumor-reactive T cells.

Shoulder pain in long-term haemodialysis patients. A clinical study of 166 patients.

We reviewed 166 adult patients on long-term haemodialysis, dividing them into three groups according to the presence and type of shoulder pain. The 24 patients in group A, with spontaneous pain related to a supine posture, had been under haemodialysis for significantly longer than the others, and had a much higher incidence of carpal tunnel syndrome. Open or arthroscopic resection of the coracoacromial ligament in 21 shoulders relieved pain during haemodialysis and night pain, and histological examination showed amyloid deposits and inflammatory-cell infiltration in the subacromial bursa in almost all cases, and in the tenosynovium of the bicipital groove in some. We conclude that one type of shoulder pain experienced by patients on long-term haemodialysis is caused by the subacromial impingement of amyloid deposits. This should be distinguished from other types of shoulder pain, because it can be relieved by resection of the coracoacromial ligament.

The prognostic importance of nuclear morphometry and the MIB-1 index in rhabdomyosarcoma [corrected].

Both nuclear morphometry and an evaluation of the cell proliferative activity have been reported to be useful tools in predicting prognosis in malignant tumors. There have been few reports, however, on rhabdomyosarcoma regarding these evaluative techniques. We performed nuclear morphometry on 51 tumor specimens obtained either by biopsy or by resection of primary and untreated rhabdomyosarcomas, and we then evaluated MIB-1 staining on 25 of the 51 specimens. The morphometric analysis was semiautomatically performed on hematoxylin- and eosin-stained sections by using a personal computer. The areas, perimeters, and lengths of the major and minor axes of the best fitting ellipse of the nuclei were all measured. In addition, the Form Ell (nuclear ellipsoidity score: minor axis/major axis) was calculated. Using the monoclonal MIB-1 antibody, which detects Ki-67 antigen in formalin-fixed, paraffin-embedded tissue, immunohistochemical studies were performed. The values of the major and minor axes and the Form Ell showed significant differences between each histologic subtype whereas the values of nuclear areas did not. We defined the nucleus whose Form ELL was smaller than 0.25 as spindle-shaped nuclei and then divided all cases into two groups on the basis of the ratio of the spindle-shaped nuclei to the total number of nuclei of the tumor cells. The group with the higher ratio showed a significantly better prognosis (P < 0.05). The same trends were also found in the specimens with an embryonal subtype. Nuclear morphometry was a useful tool in predicting prognosis. In particular, the appearance of spindle tumor cells in rhabdomyosarcoma correlated with a better prognosis.

Yolk sac tumor in a case of testicular feminization syndrome.

A 17 month old who had been diagnosed as having testicular feminization syndrome (noted during inguinal herniorrhaphy) was operated on because of an abdominal mass that had a high serum level of alpha-fetoprotein. Histologically, the lesion was a yolk sac tumor. The alpha-fetoprotein level normalized within 2 months of the surgery, through the administration of adjunctive chemotherapy containing cisplatinum. The patient is disease-free 4 years postoperatively. When performing inguinal herniorrhaphy in a girl, the surgeon should be prepared to deal with testicular feminization syndrome. If gonadal neoplasm is deniable at the time of diagnosis, careful follow-up examinations are needed until completion of the development of secondary sex characteristics.