Microcystin degradation in sphingopyxis sp. C-1

The microcystin-degrading gene cluster, mlrA-B-C-D, plaies an important role in the degradation process of hepatotoxic microcystins for several bacterial species. However after microcystin is degraded to linear-microcystin by MlrA, it is still unknown about where and by what it is metabolited. In order to clarify it, we disrupted the mlrB gene and mlrC gene in chromosome of microcystin-degrading bacteria, Sphingopyxis sp. C-1. The cells disrupted mlrB gene and mlrC gene accumulated of microcystin-degradation product, linear-microcystin and tetrapeptide, respectively, whereas the cell free extracts of ?mlrB cells detected Adda and ?mlrC cells accumulated tetrapeptide. Moreover, topology analysis of MlrB using the ß-lactamase gene fusion method insisted MlrB is the peripheral protein binding the inner-membrane. These results insist that MlrB degrades the linear microcystin in the periplasmic space and MlrC degrades tetrapeptide in cytoplasm. Thus, in intact cells, MlrC cannot degrade linear-microcystin as being separated in inner-membrane from linear-microcystin while MlrC is capable of degrading the linear-microcystin in cell-free extract

Malignant Mesodermal Mixed Tumor of the Bladder : A Case Report

We report an autopsy case of malignant mesodermal mixed tumor of the urinary bladder of a 67 year old male. He had been admitted to Kita Hospital under the diagnosis of Alzheimer\u27s disease since 1979. His dementia aggravated and he was confined to bed in 1985. While in the hospital, he developed hematuria and calculi and necrotic materials were sometimes noticed in his urine. Although the CT scan and intravenous urography revealed left hydronephrosis and vesical lithiasis, urinary cytological studies showed no remarkable changes. At the beginning of September 1992, he was found to have a child\u27s fist-sized tumor in his left lower abdominal cavity, which was diagnosed as a bladder tumor and left renal pelvic tumor. The tumor grew rapidly and reached a child\u27s head-size within 3 months. Several cytological examinations of his urine were performed, but all showed negative for malignancy. There was no tumor response against anti-cancer chemotherapy and he died on November 22, 1992. Autopsy revealed a bulky tumor mass which occupied the whole lower abdominal cavity. Histologically, the tumor was composed of an epithelial element of well differentiated squamous cell carcinoma and a non-epithelial element of rhabdomyosarcoma. The cross striations in rhabdomyosarcoma cells were clearly stained by PTAH. Immunohistochemically, rhabdomyosarcoma cells were positively stained by Desmin and partially by Vimentin. Therefore we diagnosed this tumor as malignant mesodermal mixed tumor of the urinary bladder

Incidence of endocrine-related immune-related adverse events in Japanese subjects with various types of cancer

BackgroundImmune checkpoint inhibitors (ICIs), such as cytotoxic T lymphocyte antigen-4 (CTLA-4) inhibitors, programmed cell death protein 1 (PD-1) inhibitors, and programmed cell death protein 1 ligand 1 (PD-L1) inhibitors, are often used to treat a variety of malignancies. ICIs are known to cause endocrine-related immune-related adverse events (irAEs), but the incidence varies among reports and/or agents. This study evaluated the incidence of endocrine-related irAEs in patients who were treated with ICIs in Japan.MethodThis single-center, retrospective, observational study examined the incidence and clinical characteristics of endocrine-related irAEs in 466 participants who were treated with ICIs at Kawasaki Medical School Hospital.ResultThe mean age of participants with and without endocrine-related irAEs was 69.1 ± 1.8 years and 68.1 ± 1.1 years, respectively, with no difference between them. The overall incidence of any endocrine-related irAEs among the participants was 25.5%. Hypothyroidism was prevalent in 24.3%, hypoadrenocorticism in 3.2%, hypopituitarism in 0.9%, and insulin-dependent diabetes mellitus in 1.1%. Participants receiving combination therapy with CTLA-4 and PD-1 inhibitors had a significantly higher incidence of endocrine-related irAEs than those receiving monotherapy.ConclusionEndocrine-related irAEs correlated significantly with survival and mean observation period. There was substantial difference in the incidence of endocrine-related irAEs among various types of ICIs and types of cancer. We should bear in mind that endocrine testing is necessary during the treatment with ICIs

CCR6+ MCAM+ Th17 Cell Numbers Increase in Patients with Psoriasis and Correlate with Disease Severity

Psoriasis is a chronic immune-mediated disease in which the interleukin (IL)-23/IL-17 axis plays a key role in the inflammatory cascade. We recently reported that co-expression of CCR6 and melanoma cell adhesion molecule (MCAM) in effector memory CD4 T cells (TEM) of peripheral blood might be a useful marker for T helper (Th) 17 cells. In this study, we monitored changes in TEM expressing CCR6 and MCAM using the Psoriasis Area and Severity Index (PASI) score during anti-tumor necrosis factor (TNF) therapy. We also studied CCR6+ MCAM+ Th17 cells histologically in skin biopsy samples from psoriasis patients. In psoriasis patients treated with anti-TNF therapy, the PASI score and the percentage of CCR6+ MCAM+ TEM cells in the blood changed almost in parallel. In immunohistochemical analyses, the proportions of IL-17+ T cells and MCAM+ T cells in the lesional skin of severely psoriatic patients were significantly higher than in mildly psoriatic patients (P<0.05), and the number of IL17+ T cells correlated with the PASI score (r=0.400, P<0.05). Taken together, these results indicate that CCR6+ MCAM+ Th17 cells in peripheral blood and lesional skin might play an important role in the pathology of psoriasis

Multiple transcripts of Ca 2ϩ channel ␣ 1 -subunits and a novel spliced variant of the ␣ 1C -subunit in rat ductus arteriosus

3 H]thymidine incorporation, suggesting that L-and T-type Ca 2ϩ channels are involved in smooth muscle cell proliferation in the DA. Third, we found that a novel alternatively spliced variant of the ␣ 1C-isoform was highly expressed in the neointimal cushion of the DA, where proliferating and migrating smooth muscle cells are abundant. The basic channel properties of the spliced variant did not differ from those of the conventional ␣1C-subunit. We conclude that multiple VDCC subunits were identified in the DA, and, in particular, ␣ 1C-and ␣1G-subunits were predominant in the DA. A novel spliced variant of the ␣1C-subunit gene may play a distinct role in neointimal cushion formation in the DA. alternative spliced; development; gene expression; fetal circulation THE DUCTUS ARTERIOSUS (DA) is a fetal arterial connection between the pulmonary artery and the descending aorta. After birth, the DA closes immediately, in accordance with its smooth muscle contraction. An increase in oxygen tension and a dramatic decline in circulating prostaglandins are the most important triggers of DA contraction (5). Generally, vascular smooth muscle contraction is induced by Ca 2ϩ / calmodulin-dependent phosphorylation of the regulatory myosin light chain, which is mediated by an increase in intracellular Ca 2ϩ . Ca 2ϩ influx through voltage-dependent Ca 2ϩ channels (VDCCs) and Ca 2ϩ release from intracellular stores are major sources of this increase (8, 26). Thus VDCCs must play an important role in vascular myogenic reactivity and tone of the DA. VDCCs are classified, according to their distinct electrophysiological and pharmacological properties, into low (Ttype) and high (L-, N-, P-, Q-, and R-type) VDCCs (20, In addition to their role in determining contractile state, a growing body of evidence has demonstrated that VDCCs play an important role in regulating differentiation and remodeling of vascular smooth muscle cells (SMCs) (14, In the present study, we identified multiple VDCC subunits in the DA by semiquantitative and quantitative RT-PCR and immunodetection. In particular, ␣ 1C -and ␣ 1G -subunits were predominant in the DA. Furthermore, we will demonstrate the identification of a novel spliced variant of the ␣ 1C -subunit gene that may play a role in neointimal cushion formation of the DA

Long-term follow-up on the use of vascularized fibular graft for the treatment of congenital pseudarthrosis of the tibia

<p>Abstract</p> <p>Background</p> <p>Congenital pseudoarthrosis of the tibia (CPT) is one of the most difficult conditions to treat.</p> <p>Methods</p> <p>Five girls and 3 boys with CPT were treated by vascularized fibular grafting (VFG). The average age at VFG was 7.0 years (range: 1.9–11.5 years) with an average follow-up term of 11.7 years (range: 4.9–19.6 years). Five of the children had undergone multiple operations before VFG, while the other 3 had no such history.</p> <p>Results</p> <p>Bone consolidation was obtained in all cases after an average term of 6.6 months (range: 4–10 months); this was with the first VFG in 7 cases but with the second VFG in 1 case. Complication of stress fracture and ankle pain occurred in 1 and 3 cases, respectively, only in cases undergoing multiple operations. Leg-length discrepancy was more prominent in the patients with multiple previous operations (mean: 7.5 cm), than in the cases with no prior surgery (mean: 0.7 cm).</p> <p>Conclusion</p> <p>The long-term results of VFG for CPT were excellent, especially in the cases, with no prior surgery. VFG should be considered as a primary treatment option for CPT.</p

Generation and Characterization of Conditional Heparin-Binding EGF-Like Growth Factor Knockout Mice

Recently, neurotrophic factors and cytokines have been shown to be associated in psychiatric disorders, such as schizophrenia, bipolar disorder, and depression. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the EGF family, serves as a neurotrophic molecular and plays a significant role in the brain. We generated mice in which HB-EGF activity is disrupted specifically in the ventral forebrain. These knockout mice showed (a) behavioral abnormalities similar to those described in psychiatric disorders, which were ameliorated by typical or atypical antipsychotics, (b) altered dopamine and serotonin levels in the brain, (c) decreases in spine density in neurons of the prefrontal cortex, (d) reductions in the protein levels of the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor and post-synaptic protein-95 (PSD-95), (e) decreases in the EGF receptor, and in the calcium/calmodulin-dependent protein kinase II (CaMK II) signal cascade. These results suggest the alterations affecting HB-EGF signaling could comprise a contributing factor in psychiatric disorder