Lymphotoxin production by regional lymph node lymphocytes in patients with uterine cervical cancer

The cytotoxin production by regional lymph node cells was examined in 25 patients with uterine cervical cancer and 10 patients with uterine myoma. The patients in stage I had significantly increased spontaneous release of cytotoxins compared with that in stages II, III, and IV. The spontaneous release in stages III and IV was markedly reduced. There was no difference in the release of cytotoxins from peripheral blood lymphocytes between cancer patients and patients with myoma or healthy controls. The cytotoxin production by lymph node cells was increased in stage III by stimulating with formalin-fixed QG-K cells derived from uterine cervical cancer, but not in stages I and II. Almost all of the cytotoxic activity of cytotoxin was abrogated by antilymphotoxin antibody. However, the cytotoxin activity was partially inhibited by anti-tumor necrosis factor antibody. These results suggest that cytotoxins released from the regional lymph node cells of uterine cancer patients are derived from, most of all, lymphotoxin.     

Effects of separate application of three growth factors (TGF-beta1, EGF, and PDGF-BB) on mechanical properties of the in situ frozen-thawed anterior cruciate ligament

OBJECTIVE: To clarify effects of a separate application of TGF-beta1, EGF, and PDGF-BB on the material properties of the in situ frozen-thawed anterior cruciate ligament. DESIGN: Twenty-eight rabbits were divided into four groups after undergoing the in situ freeze-thaw treatment in the right anterior cruciate ligament. In 3 of the 4 groups, 4 ng TGF-beta1, 20 ng EGF, and 4 microg PDGF-BB was applied to the frozen anterior cruciate ligament, respectively. In the remaining sham treatment group, only fibrin sealant as a vehicle was applied. Each animal was sacrificed at 12 weeks after surgery. BACKGROUND: If the role of growth factors in ligament healing and remodeling is understood, better therapies can be designed for ligament trauma. METHODS: The freeze-thaw treatment was performed three times using the originally developed cryo-probe. The cross-sectional area of the anterior cruciate ligament was measured by the optical non-contact method. After preconditioning, each specimen was stretched to failure. The ligament strain was determined with a video dimension analyzer. RESULTS: The tensile strength and the tangent modulus of the anterior cruciate ligament in the TGF-beta1 group was significantly higher than in the sham group, but significantly lower than in the normal control group. There were no significant differences in the strength and the modulus between the EGF group, the PDGF-BB group, and the sham group. CONCLUSIONS: In this model, an application of 4 ng TGF-beta1 significantly inhibited some of the material deterioration that occurs in the in situ frozen-thawed anterior cruciate ligament, while an application of 20 ng EGF or 4 microg PDGF-BB did not significantly affect the deterioration. RELEVANCE: This information will be useful in the future to develop a new biological therapy for ligament reconstruction to prevent the graft deterioration after transplantation.     

Growth autonomy and tumorigenicity of interleukin 6-dependent B cells transfected with interleukin 6 cDNA

We introduced an IL-6 cDNA expression vector into a murine B cell line, the growth of which definitely required the presence of exogenous IL-6. The transfected cells secreted substantial amounts of IL-6, to which they themselves responded by proliferating without further requirement of exogenous IL-6. The proliferation was a direct function of cell density and was inhibitable by antibodies to IL-6, indicating the autocrine nature of the growth. The IL-6 cDNA-transfected cells displayed greatly enhanced tumorigenicity when inoculated into syngeneic and nude mice. Our data suggest that an IL-6 autocrine self stimulation confers on B cells a selective growth advantage and results in the induction of progression of the malignant state of B cells.     

In vitro activity of pazufloxacin, tosufloxacin and other quinolones against Legionella species

OBJECTIVES: The activities of pazufloxacin and tosufloxacin against Legionella spp. were evaluated in vitro and compared with those of other quinolones, macrolides and azithromycin. METHODS: The conventional MICs were determined by the microbroth dilution method. Intracellular activities of drugs were evaluated by a cfu count. The minimal extracellular concentration inhibiting intracellular growth of bacteria (MIEC) was determined by a colorimetric cytopathic assay. RESULTS: MICs of pazuloxacin and tosufloxacin at which 90% (MIC90) of isolates are inhibited in 76 different Legionella spp. strains (38 ATCC strains and 38 clinical isolates) were 0.032 and 0.016 mg/L, whereas the MIC90s of levofloxacin, ciprofloxacin, garenoxacin, erythromycin, clarithromycin and azithromycin were 0.032, 0.032, 0.032, 2.0, 0.125 and 2.0 mg/L, respectively. Pazufloxacin and tosufloxacin at 4x MIC inhibited intracellular growth of Legionella pneumophila SG1 (80-045 strain), as did other quinolones, clarithromycin and azithromycin, whereas erythromycin at 4x MIC did not. MIECs of pazufloxacin, tosufloxacin, levofloxacin, ciprofloxacin and garenoxacin for the strain were 0.063, 0.004, 0.016, 0.032 and 0.008 mg/L respectively, which were superior to those of macrolides and azithromycin. Pazufloxacin showed potent activity against three additional clinical isolates of L. pneumophila SG1, one clinical isolate each of L. pneumophila SG3 and SG5, as well as Legionella micdadei, Legionella dumoffii and Legionella longbeachae SG1. CONCLUSIONS: Pazufloxacin and tosufloxacin, as well as other quinolones, were more potent than macrolides and an azalide. Present data warrant further study on the efficacy of these drugs in the treatment of Legionella infections.     

A case of pulmonary infection caused by Mycobacterium szulgai

We report a case of pulmonary non-tuberculous mycobacteriosis caused by Mycobacterium szulgai. A thirty-nine-year-old man with no relevant significant past history underwent an annual medical check. His chest X-ray and CT scan showed an infiltrative shadow with a cavity in the right upper lobe. As it was suggestive of pulmonary tuberculosis, he was referred to our hospital. Smear tests of his sputum, gastric fluid, and transbronchial fluid showed no mycobacterial organisms, but culture of the samples revealed growth of mycobacteria. The organism was identified as M. szulgai using a DNA-DNA hybridization method, and the case was diagnosed as pulmonary non-tuberculous mycobacteriosis caused by M. szulgai. By anti-mycobacterial drug treatment with isoniazid, rifampicin, and ethambutol, the infiltrative shadow on chest roentogenogram and CT showed improvement. Culture of his sputum and gastric fluid showed no growth of mycobacteria after starting treatment.     

Rigosertib induces cell death of a myelodysplastic syndrome‐derived cell line by DNA damage‐induced G2/M arrest

A multi‐kinase inhibitor, rigosertib (ON 01910.Na) has recently been highlighted as a novel type of anti‐cancer agent for the treatment of the myelodysplastic syndromes (MDS), but its action mechanisms remain to be clarified. We investigated the in vitro effects of rigosertib on an MDS‐derived cell line MDS‐L and a myeloid leukemia cell line HL‐60. Rigosertib suppressed the proliferation of both HL‐60 and MDS‐L cells and induced apoptosis by inhibition of the PI3 kinase/Akt pathway. As the effects on cell cycle, rigosertib treatment promoted the phosphorylation of histone H2AX and led to the DNA damage‐induced G2/M arrest. In addition, an immunofluorescence staining study demonstrated the abnormal localization of aurora A kinase, suggesting that rigosertib causes perturbation of spindle assembly and deregulated mitotic patterns towards cell cycle arrest and apoptosis. We also found that rigosertib exerted growth inhibitory effects on two lymphoid cell lines, Jurkat and Ramos. We further examined the molecular pathways influenced by rigosertib from the gene expression profiling data of MDS‐L cells and found a possible involvement of rigosertib treatment in the upregulation of the genes related to microtubule kinetics and the downregulation of the mRNA degradation system. The gene set enrichment analysis showed the suppression of “nonsense‐mediated mRNA decay (NMD)” as the most significantly affected gene set. These data provide a new aspect and a potential utility of rigosertib for the treatment of refractory hematopoietic malignancies.     

Involvement of extracellular calcium and arachidonate in [3H] dopamine release from rat tuberoinfundibular neurons

The mechanism of [3H]dopamine [( 3H]DA) release was investigated using primary cultures of dispersed cells from the rat tuberoinfundibular region, which contains tyrosine hydroxylase (TH)-like immunoreactive neurons. The calcium ionophore A23187 at 10 nM and above caused a significant and dose-dependent increase in [3H]DA release. In the presence of 50 microM A23187, [3H]DA release was detectable within 30 s and reached a plateau in 15 min. The induction of [3H]DA release by 50 microM A23187 was abolished by lowering the extracellular calcium concentration with 2 mM EDTA. Maitotoxin, another calcium-channel activator, also increased [3H]DA release at a concentration of 50 ng/ml. Exogenous additions of 100 mIU/ml phospholipase A2 and 10 microM arachidonate caused significant release of [3H]DA. Furthermore, A23187 stimulated [3H]arachidonate release from tuberoinfundibular dopaminergic (TIDA) neurons in a dose- and time-dependent manner. These results suggest that extracellular calcium and arachidonate are involved in the process of [3H]DA release from rat TIDA neurons.     

Advanced cholangiocarcinoma in a patient with stage I primary sclerosing cholangitis

A 29‐year‐old woman presented with jaundice and fever in May 2001. Cholangiography showed multiple strictures and beading of the biliary tree, with a large stricture in the common bile duct and marked dilatation of the hilar bile ducts. Typical cholangiography findings and elevated hepatobiliary enzymes suggested primary sclerosing cholangitis (PSC). At the same time, computed tomography detected a 2‐cm tumor in the common bile duct, and angiography showed an encasement in the portal vein. Tumor markers, cytology, and biopsy were all negative for cancer. Although laparotomy showed a healthy liver and no lymph node metastasis was found, suggesting early‐stage PSC and a low likelihood of accompanying cholangiocarcinoma (CCA) reported so far, the tumor in the resected common bile duct was subsequently diagnosed as CCA. Therefore, pancreatoduodenectomy was performed combined with partial resection of the portal trunk. Histology also revealed invasion of the wall of the portal vein by cancer cells. The patient had a recurrence 5 months later and died 12 months after her operation. This is a rare case in which stage I PSC was complicated by advanced CCA.     

Coronary artery spasm discovered in thorough examination of perioperative VT in a 26-year-old Japanese male

We thoroughly examined a 26-year-old Japanese male who experienced perioperative ventricular tachycardia. After inhaling sevoflurane, his nasal cavity was soaked with 1:100,000 epinephrine and he was intubated through the nose. Junctional tachycardia occurred five minutes after intubation, changing to ventricular tachycardia. Six-time cardioversion was required to stop the ventricular tachycardia. Echocardiography immediately following the event showed diffuse hypokinesis, and an electrocardiogram showed an inversion of T waves in II, III, aVF and V4-6. Both returned to normal within a few days. Tl scintigraphy revealed a normal perfusion image. Coronary angiography showed a normal coronary, but an injection of acetylcholine induced vasospasm in the right coronary artery. Examination of left ventricular tissue yielded no specific findings. During electrophysiological tests, ventricular tachycardia could not be induced even in the presence of isoprenaline. This is a very young case to elicit vasospasm in the coronary artery with no underlying heart disease. Although the relationship between perioperative ventricular tachycardia and coronary spasm is unknown, cardiac events can occur during anesthesia in young and low-risk patients.