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The temporal changes in the fish community structure before and after a tsunami in 2011 were investigated in Funakoshi and Otsuchi bays, Iwate prefecture, on the Pacific coast of northern Japan. Fish species richness, abundance, biomass, and species composition were analyzed in relation to temporal variability of biotic and abiotic environmental conditions in two sub-intertidal seagrass Zostera marina beds. Mean seagrass shoot density decreased to

Background There has been insufficient evidence regarding a treatment strategy for patients with non-occlusive mesenteric ischemia (NOMI) due to the lack of large-scale studies. We aimed to evaluate the clinical benefit of strategic planned relaparotomy in patients with NOMI using detailed perioperative information. Methods We conducted a multicenter retrospective cohort study that included NOMI patients who underwent laparotomy. In-hospital mortality, 28-day mortality, incidence of total adverse events, ventilator-free days, and intensive care unit (ICU)–free days were compared between groups experiencing the planned and on-demand relaparotomy strategies. Analyses were performed using a multivariate mixed effects model and a propensity score matching model after adjusting for pre-operative, intra-operative, and hospital-related confounders. Results A total of 181 patients from 17 hospitals were included, of whom 107 (59.1%) were treated using the planned relaparotomy strategy. The multivariate mixed effects regression model indicated no significant differences for in-hospital mortality (61 patients [57.0%] in the planned relaparotomy group vs. 28 patients [37.8%] in the on-demand relaparotomy group; adjusted odds ratio [95% confidence interval] = 1.94 [0.78–4.80]), as well as in 28-day mortality, adverse events, and ICU-free days. Significant reduction in ventilator-free days was observed in the planned relaparotomy group. Propensity score matching analysis of 61 matched pairs with comparable patient severity did not show superiority of the planned relaparotomy strategy. Conclusions The planned relaparotomy strategy, compared with on-demand relaparotomy strategy, did not show clinical benefits after the initial surgery of patients with NOMI. Further studies estimating potential subpopulations who may benefit from this strategy are required. Electronic supplementary material The online version of this article (10.1007/s11605-020-04792-3) contains supplementary material, which is available to authorized users.

Highlights • Retroperitoneal perforation of the colon must be considered in patients with acute pancreatitis. • The most common site of perforation is the transverse and descending colon. • Retroperitoneal drainage may lead to expansion of the perforation site, necessitating a diverting stoma., Introduction There are several reports of colon perforation in patients with acute pancreatitis, but the mechanism is not understood. We describe a patient with acute pancreatitis followed by retroperitoneal perforation of the descending colon and a duodenal fistula. Case presentation A 51-year-old male presented with acute pancreatitis. He was hemodynamically unstable, had respiratory failure on admission and was treated in the intensive care unit (ICU). He recovered and left the ICU on day 13. Although his general condition improved, a computed tomography (CT) scan showed air and fluid in the left retroperitoneum. Gastrografin enema and CT scan showed extraluminal leakage in the descending colon and retroperitoneal drainage was performed on day 27. After drainage, there was continuous voluminous feculent discharge, and a loop ileostomy was performed on day 34. A repeat CT scan revealed ascites. A percutaneous catheter injected with contrast showed a duodenal fistula. After drainage, the fever resolved and the patient was discharged on hospital day 106. Discussion Although there is no clear mechanism of colonic perforation in patients with acute pancreatitis, one hypothesis is that ischemia secondary to inflammation caused by pancreatitis plays a role. The involved area is usually in the watershed areas of the colon. Retroperitoneal drainage of the colon perforation may have necessitated creation of a diverting loop ileostomy. Conclusion Retroperitoneal colon perforation must be considered in patients with acute pancreatitis.

Highlights • Manual reduction was successfully performed for a patient with sciatic hernia. • There were no report of closed manual reduction previously. • A sciatic hernia in women may be manually reduced without surgery., Introduction Sciatic hernias are the least common type of pelvic floor hernias. The purpose of this study was to present a novel technique for manual reduction and to conduct a systematic review of previous reports of sciatic hernias to characterize them and review the outcomes. Presentation of case An 86-year-old female presented with left-sided lumbar pain. She had a past medical history of rheumatoid arthritis and was treated with prednisolone and methotrexate. Her left abdomen and left lumbar area were tender. An unenhanced abdominal computed tomography scan revealed invagination of the left ureter into the left sciatic foramen and a dilated left proximal ureter and renal pelvis. Ultrasonography showed an invaginated left ureter viewing from the left buttock. She was diagnosed with a sciatic hernia. Ultrasound-guided manual transvaginal reduction was performed. Post-procedure unenhanced abdominal computed tomography scan confirmed reduction of the ureter. After 10-months of follow-up, there is no evidence of recurrence. Discussion Previous reports of patients with sciatic hernia were identified. Clinical data associated with the hernia, reduction technique and clinical outcomes were collected for 72 patients. Open reduction was performed in 24 patients. A ureteral stent was placed in eight patients when the hernia contained the ureter. Four postoperative complications including one death were reported in adults. There were no reports of closed manual reduction. Conclusion A sciatic hernia in women may be manually reduced without surgery. Further reviews of this rare entity are needed to determine the best management strategy.

Background There are few reports of metastases from colon cancer to an inguinal hernia sac, and few reports of colon cancer originating in diverticula. We report a patient with carcinoma of the sigmoid colon arising in two diverticula, who presented with peritoneal seeding to an inguinal hernia sac, and a review of the literature. Case presentation A 55-year-old male underwent open herniorrhaphy for a left inguinal hernia. At operation, a nodule in the inguinal hernia sac was resected and histologic examination revealed adenocarcinoma, which was suspected to be a metastasis from a distant primary lesion. Postoperative evaluation included colonoscopy and positron emission tomography which showed two suspected lesions in sigmoid diverticula. Laparoscopic subtotal colectomy was performed, and pathology revealed adenocarcinoma in two sigmoid diverticula. Conclusions If a nodule is found in an inguinal hernia sac, especially in older patients, peritoneal metastases should be considered. Resection of the nodule with histopathologic evaluation is essential. Colon cancer arising in a diverticulum should be considered as a possible site of the primary lesion.

Historically defined in the European Group for the Immunological Characterization of Leukemias (EGIL) [1], acute bilineal leukemia (ABLL) is characterized by 2 distinct blast populations (e.g. B/my...

Telemedicine for chronic disease management is extending to the home through the use of medical devices and ICT technologies. Patients with intractable diseases, such as amyotrophic lateral sclerosis (ALS) and lethal neurodegenerative diseases, have been returning to their homes rather than remaining hospitalized. Reliable alarms for condition changes of patients and burden reduction of their families are taking root as foundations of telemedicine for patients with intractable diseases. This paper discusses reliable alarm delivery and expected medical IoT features for those patients. A patient's family has difficulty in setting optimal parameters of life-support medical devices following patient condition changes. Also, caregivers and patients' families expect reliable alarms and false alarm reduction from tele-alarm systems used at home. We need to provide both anxiety relief for patients' families and patient safety by reliably monitoring the patients. We designed and implemented an alarm delivery system for patients with intractable diseases, and here we propose a prototype false-alarm reduction mechanism for highly-controlled medical device systems including an artificial ventilator. We investigated alarms of a patient for one year, cooperating with the patient's family. We need both hardware standard interfaces and consistent alarm functions between artificial ventilators. We conclude with our further work for patients with different types of intractable diseases and for standardization of medical IoT networks integrating false-alarm reduction systems.

To investigate the response of the fish community structure to a natural disturbance in their habitat, fish abundance, biomass and species composition were analysed in relation to temporal variability of environmental conditions in a seagrass Zostera marina bed. A total of 3024 fishes belonging to 46 taxa (22 families) were collected by quantitative sampling for 10 years from 2007 to 2016 in the Seto Inland Sea, south-western Japan. Seagrass shoot density decreased to less than 1/20 of its original density after disappearance of vegetation caused by heavy rain in the autumn of 2011 and the area did not recover for the next five years. In order to analyse temporal changes of fish community, the fishes were divided into three groups depending on their habitats or lifestyles: pelagic or migratory species (PM), sand or mud bottom-associated species (SM) and seagrass (Z. marina) – or substrate (rocky bottom including macrophytes) – associated species (ZS). Multiple regression analysis showed seagrass shoot density had the most significant effect on biomass of ZS among the three groups, with higher fish biomass under higher seagrass shoot density. Fish community composition changed after the disappearance of the seagrass vegetation coverage with an increase in abundance of SM during the five years of the post-disturbance period. Seagrass vegetation was concluded to affect temporal change of fish community structure through a stronger influence on fish species that are more dependent on seagrass beds as habitat., This study was partially supported by the Global Environment Research Fund (J.S., grant number RF-0907and RF-1102) of the Ministry of the Environment, and Grant-in-Aid for Scientific Research (B) (J.S., grant number 24380107).

Highlights • 75% of pancreas transplantation is SPKT. • The optimal surgical management for graft perforation is still unknown. • Delayed graft duodenal perforation is very rare. • The omega loop may contribute to increased pressure by luminal congestion. • When delayed duodenal graft perforation occurs, graft excision may not be necessary., Introduction Pancreas transplantation is the best treatment option in selected patients with type 1 diabetes mellitus. Here we report a patient with a nonmarginal duodenal perforation five years after a simultaneous pancreas-living donor kidney transplantation (SPLKT). Presentation of case A 31-year old male who underwent SPLKT five years previously presented with severe abdominal pain. He had a marginal duodenal perforation four years later, treated by primary closure and drainage. Biopsy of the pancreas and duodenum graft at that time showed chronic rejection in the pancreas and acute inflammation with an ulcer in the duodenum. At presentation, computerized tomography scan showed mesenteric pneumatosis with enteric leak and ileal dilatation proximal to the anastomotic site. We performed emergent laparotomy and found a 1.0 cm perforation at the nonmarginal, posterior wall of the duodenum. Undigested fiber-rich food was extracted from the site and an omental patch placed over the perforation. An ileostomy was created proximal to the omega loop for decompression and a drain placed nearby. The postoperative course was unremarkable. Discussion There are only eight previous cases of graft duodenal perforation in the literature. Fiber-rich food residue passing through the anastomosis with impaction may have led to this perforation. Conclusion When a patient is stable, even in the presence of delayed duodenal graft perforation, graft excision may not be necessary. Intraoperative exploration should include Doppler ultrasound examination of the vasculature to rule out thrombosis as a contributor to ischemia. Tissue biopsy should be performed to diagnose rejection.

A 15-year-old girl with acute lymphoblastic leukemia received allogeneic dendritic cell vaccination, pulsed with Wilms tumor 1 (WT1) peptide, after her third hematopoietic stem cell transplantation (HSCT). The vaccines were generated from the third HSCT donor, who was her younger sister, age 12 years. The patient received 14 vaccines and had no graft-versus-host disease or systemic adverse effect, aside from grade 2 skin reaction at the injection site. WT1-specific immune responses were detected after vaccination by both WT1-tetramer analysis and enzyme-linked immunosorbent spot assay. This strategy may be safe, tolerable and even feasible for patients with a relapse after HSCT.

Long-term surviving recipients of allogeneic hematopoietic stem cell transplantation (HSCT) often suffer from diabetes mellitus (DM). We sought to identify risk factors for the development of post-transplant DM and impaired glucose tolerance (IGT) in pediatric HSCT patients. Glucose tolerance statuses were evaluated in 22 patients aged 6.3-21.8 years who had received allogeneic HSCT between the ages of 0.8-13.5 years. Five patients were diagnosed as having type 2 DM, and treated with insulin or oral hypoglycemic agents. Five patients were included in the IGT group, and the remaining 12 children were in the normal glucose tolerance (NGT) group. The cumulative incidence of DM plus IGT was 11.6 % at 5 years and 69.3 % at 10 years. None of the patients were obese/overweight and none had a family history of DM. There were no significant differences in serum levels of leptin and adiponectin between the DM + IGT and the NGT groups. An average preprandial glucose levels in the DM + IGT group were significantly higher than those in the NGT group from preparative conditioning to 60 days after HSCT. In multivariate analysis, an age of ≥6 years at the time of HSCT was significantly associated with the development of DM + IGT. Additionally, careful follow-up is necessary, even for NGT patients.

advance online publication, February 25, 2014, Juvenile myelomonocytic leukemia (JMML) is a fatal, mixed myeloproliferative and myelodysplastic disorder occurring in infancy and early childhood. Children with JMML have mutually exclusive genetic abnormalities in granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling pathways, inactivation of the NF1 or mutations in PTPN11, NRAS, KRAS and CBL. A whole-exome sequencing study, performed by Sakaguchi et al.,3 has recently demonstrated that in addition to the high frequency of RAS pathway mutations, mutations in SETBP1 and JAK3 are common recurrent secondary events, and that these events may be involved in tumor progression, and are associated with poor clinical outcomes., Article, LEUKEMIA. 28(7):1545-1548 (2014)

Afferent loop syndrome is a rare complication of gastric surgery. An obstruction of the afferent limb can present in various ways. A 73-year-old man presented with one day of persistent abdominal pain, gradually radiating to the back. He had a history of total gastrectomy with a Roux-en-Y reconstruction. Abdominal computed tomography scan revealed dilation of the duodenum and small intestine in the left upper quadrant. Exploratory laparotomy showed volvulus of the biliopancreatic limb that caused afferent loop syndrome. In this patient, the 50 cm long limb was the cause of volvulus. It is important to fashion a Roux-limb of appropriate length to prevent this complication.

Background: Chronic granulomatous disease (CGD) is a rare inherited primary immunodeficiency that affects phagocytic cells. CGD patients are susceptible to fungal infections, especially Aspergillus infections. The management of life-threatening Aspergillus infections in CGD is particularly difficult because some infections cannot be eradicated with standard antifungal treatments and, hence, result in death. Case Report: A 2-week-old girl developed invasive pulmonary aspergillosis, which rapidly progressed to respiratory failure. Liposomal amphotericin B, micafungin, and voriconazole were not effective. At the age of 2 months, she was diagnosed with p67phox-deficient CGD. In addition to antifungal treatment, the patient received 21 granulocyte transfusions (GTX), which were obtained from 300- or 400-mL whole blood samples from healthy random donors who were not treated with granulocyte–colony-stimulating factor or dexamethasone. The median neutrophil count of the GTX was 1.88 × 108/kg body weight. Rituximab was administered to reduce alloimmunization to human leukocyte antigens (HLA) after the eighth GTX, resulting in their absence of anti-HLA before and after cord blood transplantation (CBT). A marked improvement in her invasive pulmonary aspergillosis was achieved, although the first CBT was rejected. Complete hematopoietic recovery was obtained after the second CBT. Conclusion: Repeated GTX containing relatively low doses of neutrophils might be able to control severe Aspergillus infections in infants with CGD.

Background Allergic transfusion reactions (ATRs), particularly those caused by plasma-rich platelet concentrates (P-PCs), are an important concern in transfusion medicine. Replacing P-PCs with PCs containing M-sol (M-sol-R-PCs) is expected to prevent ATRs. However, this has not yet been verified by sufficient clinical evidence. Study Design and Methods A retrospective cohort study was performed between 2008 and 2011. Pediatric patients with hematologic disorders, solid tumors, primary immunodeficiency disorders, or inherited metabolic disorders were transfused with M-sol-R-PCs between 2010 and 2011; the transfusions of P-PCs administered between 2008 and 2011 were compared in terms of frequency and severity of ATRs, corrected count increment (CCI), and occurrence of bleeding. Data were collected for 6 consecutive months on a per-patient basis. Results Data obtained during 2008 to 2011 showed that of the 78 patients receiving 515 P-PC transfusions, 14 (17.9%) had 17 ATRs (3.3%); 14 and three ATRs were of Grades 1 and 2, respectively. In 2010 to 2011, 49 patients received 620 transfusions of M-sol-R-PCs, and two patients (4.1%) had Grade 1 ATRs (0.3%). Thus, the frequency of ATRs per bag and per patient differed significantly between the two transfusions. No steroid agents were used for the prevention or treatment of ATRs in the M-sol-R-PC group. The CCI (24 hr) for M-sol-R-PCs did not differ from that for P-PCs. Conclusion M-sol-R-PCs were found to be effective in preventing ATRs without loss of transfusion efficiency in children; however, its efficacy should be further evaluated in prospective clinical trials.

There have been no reports of human herpesvirus-6 (HHV-6) encephalitis treatment based on both HHV-6 DNA load and the antiviral agent's concentration in the cerebrospinal fluid (CSF).A 20-year-old male with a hematological malignancy developed HHV-6 encephalitis 15 days after unrelated cord blood transplantation (UCBT). He had fever, chest pain, memory impairment, and insomnia. His CSF showed no increased cell counts, but the amount of HHV-6 DNA was elevated to 2.0 × 10(6) copies/ìgDNA. Magnetic resonance imaging (MRI) of the head revealed abnormal high-intensity signals in the left limbic system on T2-weighted and diffusion-weighted images. Intravenous administration of ganciclovir (GCV) was initiated at 5 mg/kg every 12 h on day 18, and was continued until day 137. The amount of HHV-6 DNA in the plasma became undetectable on day 25. The HHV-6 load in the CSF decreased to 1.5 × 10(3) copies/ìgDNA on day 32, and reached undetectable levels on day 53. The mean concentration of GCV 1 h after an infusion of 5 mg/kg was 4.12 mg/mL in plasma and 0.7 mg/mL in CSF. The chest pain and insomnia disappeared on days 35 and 47, respectively. Memory defects recovered up to day 85.Serial quantification of HHV-6 DNA in CSF may be useful for successful treatment with GCV in post-transplant HHV-6 encephalitis.

Preperitoneal hematomas are rare and can develop after surgery or trauma. A 74-year-old woman, receiving systemic anticoagulation, developed a massive preperitoneal hematoma after a subcutaneous injection of teriparatide using a 32-gauge, 4 mm needle. In this patient, there were two factors, the subcutaneous injection of teriparatide and systemic anticoagulation, associated with development of the hematoma. These two factors are especially significant, because they are widely used clinically. Although extremely rare, physicians must consider this potentially life-threatening complication after subcutaneous injections, especially in patients receiving anticoagulation.

Corn fiber (CNF) is an abundant by-product of the wet corn milling process in the production of cornstarch. We have shown that the hot water-soluble fraction (HWSF) from CNF has a promoting effect on the mycelial growth of various edible mushrooms, including mycorrhizal fungi. To reveal the promoting mechanisms, the effect of CNF-HWSF on the stimulation of extracellular enzymes was examined. The production of extracellular carbohydrases such as amylase, CMCase, and xylanase was markedly enhanced by the addition of low molecular weight fractions (less than MW 500) prepared from CNF-HWSF. The enzymatic stimulation and enhancement of mycelial growth appeared during 3–15 days after inoculation. Furthermore, a fraction of less than MW 500 was separated by gel filtrate chromatography into five fractions (A–E), and the effect of each fraction was investigated. Promoting effects were shown from C and D fractions; mycelial growth and enzyme production of Lentinula edodes were indicated although fraction D has no sugars and amino acids in CNF-HWSF. From these results, the promoting effect of CNF-HWSF seems to be a two-step reaction. The first step could be achieved by rich nutrients such as free amino acids and monosaccharides from CNF-HWSF. The second step (during 3–15 days) is considered to be that the marked promoting effect was caused by the stimulation of extracellular enzymes.

No standard treatment has been established in childhood blastic plasmacytoid dendritic cell neoplasma (BPDCN) because of its rarity. We treated with acute lymphoblastic leukemia-type regimen for a child with BPDCN with skin and leukemic involvement. She has been disease-free for 4 years after allogeneic bone marrow transplantation in first complete remission. In 33 cases of pediatric BPDCN, the over survival was significantly lower in the patients with skin manifestation than those without cutaneous involvement. Accordingly, it is important to determine whether allogeneic hematopoietic stem cell transplantation should be applied to first complete remission in the patients with poor prognosis.

市販梅酒 (アルコール14%, 糖20%, エキス分30%含有) 100mlを毎日6カ月間継続飲用させた10人(43.5±15.2歳) を被験者とし, 梅酒飲用による健康人の血中脂と血圧に及ぼす効果についての予備的研究を行った。毎月1回, 身長, 体重, 血圧, 検尿 (尿蛋白質, 尿糖, ウロビリノーゲン, ケトン体) 測定を行った。原則として空腹時採血し, 血清総コレステロール, HDL-コレステロール, 血糖値, ヘモグロビンA1C, ヘモグロビン量, アルブミンを測定した。またBMI, 動脈硬化指数は計算によって求めた。その結果, HDL-コレステロールは飲用前値59.0mg/dlであったが, 飲用2カ月後から有意に増加し, 6カ月後に64.1mg/dlになった。また動脈硬化指数は飲用前値2.54で, 飲用2カ月後から有意に低下し, その後一定値を維持した。収縮期血圧は前値132.8mmHgであったが, 6カ月後に128.7mmHgと低下傾向を示した。拡張期血圧は飲用前値88.0mmHgであったが6カ月後に80.2mmHgと有意に低下した。血糖値は6カ月間に変化は認められず, ほぼ一定の87～89mg/dlを維持した。ヘモグロビンA1Cは血糖値と同様に有意の変化は認められず, 6カ月間ほぼ一定の4.8～4.9%であった。その他の検査に有意な変動は認められなかった。

Corn fiber (CNF) is an abundant by-product of the wet corn milling process used to produce corn starch. In light of the need to recycle organic wastes, the effects of adding a hot water-soluble fraction (HWSF) from CNF to a medium on the vegetative mycelial growth of nine edible mushrooms such as Lentinula edodes and Pholiota nameko were investigated. The results showed that the mycelial growth of these fungi was markedly increased (1.4–9.5 times that of the control) by adding 5%–20% CNF-HWSF to the medium. These promotive effects were also apparent on mycorrhizal mushrooms, such as Tricholoma matsutake (3.3-fold) and Lyophyllium shimeji (3.7-fold). The promotive effects on mycelial growth were shown in the low-molecular-weight fractions (molecular weight

We describe the effects of trehalose on spawn storage in a home freezer (average temperature, −16°C) where edible fungi usually do not survive. When the mycelia of Lentinula edodes were stored in a freezer for 3 days, the survival rate of mycelia cultivated on 2% glucose medium was 30%, whereas those on media containing 2% and 5% trehalose were 50% and 60%, respectively. Addition of trehalose to the culture was more effective in Pleurotus ostreatus. These results suggest that trehalose played the role of a stress protectant against freezing, because the mycelia cultured on a trehalose medium grew more rapidly and produced more fruiting bodies compared to those cultured on glucose.

The gene that encodes a thermostable endo-arabinase (called ABN-TS) from Bacillus thermodenitrificans TS-3 was cloned, sequenced, and expressed in the mesophilic B. subtilis. The gene contained an open reading frame consists of 939 bp, which encodes 313 amino acids. The deduced amino acid sequence of the enzyme showed 50, 46, and 36% similarity with endo-arabinase from B. subtilis IFO 3134 (PPase-C), Pseudomonas fluorescens (ArbA), and Aspergillus niger (ABNA), respectively. The hydrophobic and acidic amino acids making up ABN-TS outnumbered those in PPase-C. The gene product expressed in B. subtilis, as the host, had substantially the same characteristics, and was stable up to 70 degrees C, and the reaction was optimal around 70 degrees C, as well as native ABN-TS.

Conventional myeloablative conditioning (MAC) regimens often cause severe regimen-related toxicity (RRT). Furthermore, many patients suffer from poor quality of life in accordance with the increase in long-term survivors. We therefore devised a reduced-toxicity myeloablative conditioning (RTMAC) regimen consisting of 8-Gy total body irradiation (TBI), fludarabine (FLU) and cyclophosphamide (CY) for pediatric hematological malignancies. A retrospective single-center analysis was performed on patients with leukemia or myelodysplastic syndrome (MDS), aged ≤20 years, who had received an 8-Gy TBI/FLU/CY RTMAC regimen followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT). Thirty-one patients underwent first allo-HSCT after an RTMAC regimen. The diagnoses were acute lymphoblastic leukemia (n = 11), acute myeloid leukemia (n = 13), MDS (n = 4), juvenile myelomonocytic leukemia (n = 1) and acute leukemias of ambiguous lineage (n = 2). While 3 patients showed early hematological relapse, the remaining 28 patients achieved engraftments. None of the patients developed grade 4 or 5 toxicities during the study period. The 5-year overall survival and relapse-free survival were 80% [95% confidence interval: CI, 61–91%] and 71% [95% CI, 52–84%], respectively. Our RTMAC regimen would be less toxic and offers a high probability of survival for children with hematological malignancies.

The gene that encodes a thermostable pectate lyase (called PL 47), from Bacillus sp. TS 47, was cloned, sequenced, and expressed in mesophilic B. subtilis. The gene contained an open reading frame consisting of 1326 bp, which encoded 441 amino acids. The deduced amino acid sequence of the mature enzyme (416 amino acids with a calculated molecular mass of 47,262 Da), showed 52% similarity with PL (BsPel) from mesophilic B. subtilis SO113. The structure-based alignment of the deduced amino acid sequence of PL 47 with that of BsPel suggested that PL 47 might have a parallel beta-helix structure with three long loops. The amino acids making up PL 47 are richer in hydrophobic amino acids and glutamic acid than BsPel. The hydropathy profile of PL 47 indicated that the amino acid sequences around putative calcium binding sites are more hydrophobic than the same region of BsPel. The gene product expressed in B. subtilis as the host was stable up to 70 degrees C and the reaction was optimal around 70 degrees C, as well as native PL 47.

A strain of thermophilic bacterium, Bacillus sp., with pectolytic activity has been isolated. It produced an extracellular endo-polygalacturonate trans-eliminase (PL, EC 4.2.2.1) when grown at 60 degrees C on a medium containing polygalacturonate (PGA). The PL was purified by hydrophobic, cation exchange, and size exclusion column chromatographies. The molecular mass of the enzyme was 50 kDa by SDS-PAGE. The isoelectric point of the enzyme was pH 5.3. The enzyme had a half-life of 13 and 1 h at 65 and 70 degrees C, respectively, and showed optimal activity around at 70 degrees C and pH 8.0. It had protopectinase activity, besides PL activity, on lemon protopectin and cotton fibers. The first 20 amino acids sequence of the enzyme had significant similarity with that of PL from methophilic Bacillus subtilis, with 50% identity.

In order to develop a new tester strain detecting environmental promutagens and procarcinogens, we introduced two plasmids into Salmonella typhimurium TA1535; one contains the cDNAs of human cytochrome P450 (P450 or CYP) 1A2 and NADPH-P450 reductase and the other (pOA101) a umuC"lacZ fusion gene. The newly developed tester strain, S. typhimurium OY1001/1A2, was found to express P450 at a level of 0.15 nmol/ml in whole cell culture. Membrane fractions, when isolated from this tester strain, contained 0.04 P450 nmol/mg protein and a reductase activity of 170 nmol cytochrome c reduced/min/mg protein and were active in catalyzing CYP1A2-dependent 7-ethoxyresorufin O-deethylation and metabolic activation of heterocyclic aromatic amines to DNA-damaging products in a conventional tester S. typhimurium NM2009 strain, only when NADPH was added as a reducing equivalent. In the OA1002/1A2 strain, heterocyclic aromatic amines (e.g., IQ, MeIQ, and MeIQx) were found to be activated to reactive metabolites that cause induction of umuC gene expression in a dose-dependent manner, without addition of external NADPH. These results indicate that the newly established strain can be of use to detect mutagenic and carcinogenic potencies of environmental chemicals without addition of metabolic activation system.

Purification and some properties of an isoform of metal proteinase fromHypsizygus marmoreus are described. This enzyme was purified 711-fold with 5.44% recovery. The molecular weight and pl value were 41,500 and pH 7.7, respectively. The highest activity was observed against milk casein as the substrate. This enzyme was strongly inhibited by metal proteinase inhibitors such as phosphoramidon, EDTA, ando-phenanthroline.

We investigated the possible role of metal proteinase on the fruit-body formation ofHypsizygus marmoreus. The addition of a specific metal proteinase inhibitor, phosphoramidon, to the culture medium (10μg/ml) completely inhibited fruit-body formation. Metal proteinase activity in both the medium and the mycelia of this fungus increased markedly during vegetative mycelial growth, and activity was maximal 25 days after inoculation. When phosphoramidon was added to the culture medium during vegetative mycelial growth, the metal proteinase activity in the mycelium decreased to 56% of the control (without inhibitor) level. Isoelectric focusing analysis showed that two kinds of metal proteinases with a pl of 7.7 and 8.4, respectively, were obtained from 29-day-old mycelia. Uptake of phosphoramidon into the mycelia was confirmed as the result of inhibition of thermolysin activity by the mycelial extracts. The degree of inhibitor uptake into mycelia was about 2.0% and was independent of the initial concentration of the inhibitor administered. The addition of peptone and amino acids to medium treated with phosphoramidon resulted in fruit-body dry weight yields that were about 50% that of the control.

Modifications of the standard Ames test and umu test were employed to study the “desmutagenic” response exerted by pheophytin b against the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). The mixture from which pheophytin b separated was activated with S9 mix and was assayed to determine the remaining mutagenicity of Trp-P-2. The decrease in the His+ revertant colonies of Salmonella typhimurium TA98 was dependent on the increase in pheophytin b concentrations. Binding of Trp-P-2 to DNA and the subsequent induction of the SOS response were investigated using the umu test employing Salmonella typhimurium NM 2009. Trp-P-2 induced induction of β-galactosidase was suppressed when pheophytin b was included in the test. In order to evaluate the mode of action of pheophytin b in the mixture of Trp-P-2, Trp-P-2 was analyzed using HPLC. Compared with a solution of Trp-P-2 alone, the absorbance peak of Trp-P-2 in the HPLC profile of the mixture of Trp-P-2 and pheophytin b was observed to be decreased. Also a new peak was not shown in the HPLC profile. To investigate the possible effect of pheophytin b on the S9 mix, the HPLC profile of a mixture containing Trp-P-2, S9 mix and pheophytin b was compared with a mixture containing only Trp-P-2 and S9 mix. The absorbance peak of the hydroxylated Trp-P-2 in the reaction mixture containing pheophytin b was observed to be smaller than the peak of the mixture without pheophytin b. From these results, it is suggested that pheophytin b limits the formation of hydroxylated Trp-P-2.

The effects of short periods (15-200 days) of either refrigerated or room temperature spawn storage on the mycelial growth, fruit-body yield, chemical constituents and enzyme activities of the mycelia of three strains of Lentinus edodes were investigated. No significant decrease in the mycelial growth and fruit-body yield was observed under storage for 200 days at 4°C, 160 days at 15°C or 60 days at 24°C. Mycelial growth increased upon spawn storage for 60-110 days at 4°C or 15°C. These effects were observed in all strains tested. The fruit-body yield increased on storing the spawn for 30 days at 15°C. However, storage of the spawn for relatively longer periods (160-200 days) at low temperature (4°C, 15°C) decreased the formation of morphologically normal fruit-bodies. In contrast, normal fruit-body formation increased upon storage for shorter periods at 24°C compared with the not-stored spawn (control) . Total sugar in mycelia after incubating the stored spawn decreased significantly, whereas the protein and free amino acid content did not show any decrease. Proteinase activity in mycelia after incubating the stored spawn did not differ significantly. However, amylase, β-1, 3-glucanase and trehalase activity decreased markedly upon storage of the spawn.

Micafungin (MCFG) is used for the prophylaxis of invasive fungal disease (IFD) after allogeneic hematopoietic stem cell transplantation (HSCT). However, the safety, efficacy, or optimal dosage/blood levels as prophylaxis is uncertain in pediatric HSCT-patients. We prophylactically administered MCFG at 2 mg/kg once daily to 38 children and adolescents undergoing allogeneic HSCT. During MCFG prophylaxis, infusion reactions or adverse events (grades 2–5) related to MCFG use were not found in all the patients. Thus, MCFG prophylaxis was not discontinued and other antifungal agents were not added except for 2 patients in whom probable or possible IFDs developed (completion rate, 94.7 %). To elucidate the influence of HSCT-related complications/drugs on blood concentration of MCFG, we determined the plasma trough and peak levels in 13 and 10 among 38 patients, respectively. The mean trough and peak levels were 3.04 ± 1.21 μg/mL (569 samples) and 9.63 ± 3.62 μg/mL (44 samples), respectively. The peak levels were moderately correlated to the trough levels (R 2 = 0.466). In a patient, the trough level of MCFG transiently increased up to 10.21 μg/mL during hepatic dysfunction due to acute graft-versus-host disease. The MCFG trough levels strongly correlated with T-Bil value (R 2 = 0.894). There was no relationship between the trough levels of MCFG and the circulating concentrations of tacrolimus (R 2 = 0.040). Additionally, MCFG levels were not influenced by treatment with cyclophosphamide or corticosteroids. Prophylaxis with MCFG at 2 mg/kg once daily may be safe, tolerable, and feasible in pediatric HSCT-patients.

No standard treatment has been established in childhood blastic plasmacytoid dendritic cell neoplasma (BPDCN) because of its rarity. We treated with acute lymphoblastic leukemia-type regimen for a child with BPDCN with skin and leukemic involvement. She has been disease-free for 4 years after allogeneic bone marrow transplantation in first complete remission. In 33 cases of pediatric BPDCN, the over survival was significantly lower in the patients with skin manifestation than those without cutaneous involvement. Accordingly, it is important to determine whether allogeneic hematopoietic stem cell transplantation should be applied to first complete remission in the patients with poor prognosis.

Chronic granulomatous disease (CGD) is a rare inherited primary immunodeficiency that affects phagocytic cells. CGD patients are susceptible to fungal infections, especially Aspergillus infections. The management of life-threatening Aspergillus infections in CGD is particularly difficult because some infections cannot be eradicated with standard antifungal treatments and, hence, result in death.A 2-week-old girl developed invasive pulmonary aspergillosis, which rapidly progressed to respiratory failure. Liposomal amphotericin B, micafungin, and voriconazole were not effective. At the age of 2 months, she was diagnosed with p67phox-deficient CGD. In addition to antifungal treatment, the patient received 21 granulocyte transfusions (GTX), which were obtained from 300- or 400-mL whole blood samples from healthy random donors who were not treated with granulocyte-colony-stimulating factor or dexamethasone. The median neutrophil count of the GTX was 1.88 × 10(8) /kg body weight. Rituximab was administered to reduce alloimmunization to human leukocyte antigens (HLA) after the eighth GTX, resulting in their absence of anti-HLA before and after cord blood transplantation (CBT). A marked improvement in her invasive pulmonary aspergillosis was achieved, although the first CBT was rejected. Complete hematopoietic recovery was obtained after the second CBT.Repeated GTX containing relatively low doses of neutrophils might be able to control severe Aspergillus infections in infants with CGD.

Using the Salmonella/microsome assay, the antimutagenic effects of specific components of the extracts from eggplant fruits were investigated. The eggplant fruit juice exhibited an antimutagenic activity against 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) induced mutagenicity. In some of the fractions extracted with several organic solvents (acetone, petroleum ether, ethyl acetate; and methanol), the activity was recognized. No mutagenicity or toxicity for Salmonella typhimurium TA98 in the presence of S9 mixture was observed with any of the extracts. It is suggested that there are multiple components of the activities that exist in the eggplant fruit. We isolated lutein from the 84% methanol (methanol/water, v/v) layer, pheophorbide or chlorophyllide from the 70% methanol layer and tannins containing sugar-moieties from the water layer. Pheophytin a and b, Mg-free derivatives of chlorophyll a and b, were isolated from the petroleum ether layer as possible antimutagens. The pheophytin a with S9 mix inhibited by 30-40% the mutagenicity of Trp-P-2.

To survey the potential for production of extracellular hydrolytic enzymes by mycorrhizal mushrooms, productivities of these exo-enzymes from mycelia on potato-dextrose liquid medium were determined.Tricholoma matsutake produced relatively high levels of CM-cellulase and avicelase activities in all test strains. It also produced higher activity of acid proteinase than neutral proteinase. Its xylanase activities seemed to be higher than those of the other carbohydrases. The productivities ofLyophyllum shimeji strains were at similar levels to those ofT. matsutake strains. CM-cellulase and avicelase activities ofL. shimeji were higher than those ofT. matsutake. Neutral proteinase inL. shimeji strains showed higher activity levels than acid proteinase. The relative productivities of hydrolytic enzymes between the groups of mycorrhizal mushrooms and wood-rotting mushrooms were also examined.T. matsutake andL. shimeji both produce many kinds of hydrolytic enzymes in their culture broth, and the potential for production of hydrolytic enzymes by mycorrhizal mushrooms was judged to be relatively high.

In order to explore the relationship between proteinase and fruit body growth and basidiospore formation in mushrooms, the metallo proteinase from the pilei in the fruit bodies of Enokitake (Flammulina velutipes) was purified and characterized. The metallo proteinase was highly purified (194-fold) with a recovery of 5.90%. The enzyme was the most active at pH 6.5 toward casein, and stable only within the narrow range of pH 6.0 to 8.0 even under mild temperature (37°C, 30 min). The molecular weight and isoelectric point were found to be 18,000 and pI 6.0, respectively. The enzyme was strongly inhibited by MK-I or synthetic metallo proteinase inhibitors such as ethylenediamine-tetraacetic acid and o-phenanthroline. These characteristics were compared with those of other metallo proteinases, and the possible roles of the enzyme are discussed.

きのこの子実体生育と担子胞子の形成に子実体中のプロティナーゼがどのようなかかわりを持つかを明らかにする目的で,エノキタケを培養し,生産されるプロティナーゼについて検討し,以下の結果を得た.(1) 本菌の子実体には活性至適pH 3.2と6.5を示す酸性プロティナーゼおよび中性プロティナーゼが存在した.両酵素の活性は子実体の生育に伴って,菌柄部では大きな変動はなかったが,菌傘部で著しく上昇した(約12倍).酸性プロティナーゼは過熟子実体で最高値(3.0×102U/ペトリ皿)を示し,中性プロティナーゼは成熟子実体形成時に最も高い活性(5.9×102U/ペトリ皿)を示した.菌傘部の中性プロティナーゼの総活性を成熟子実体で比較すると酸性プロティナーゼの約4倍で,比活性でも3.3倍であった.(2) 各種の酵素阻害剤に対する挙動から酸性プロティナーゼはカルボキシルタイプ,中性プロティナーゼはメタルタイプであることを確認した.また,メタルプロティナーゼのpIは6.0であった.これらの結果から,エノキタケの子実体傘部の生育には菌傘部で生産されるpI 6.0のメタルプロティナーゼが重要で,担子胞子の形成にもこの酵素が主要な働きをするものと推察された.