Effects of Pathogen Density, Soil Moisture, and Soil pH on Biological Control of Clubroot in Chinese Cabbage by Heteroconium chaetospira.

The effects of soil moisture and pH, and pathogen resting spore density, on the effectiveness of the biological control of clubroot by the fungal endophyte Heteroconium chaetospira was evaluated in greenhouse and field experiments. Conditions favoring disease development included low pH (5.5) and high soil moisture content (80%), with significant reductions in the disease being observed at a higher pH (6.3 and 7.2) and lower soil moisture content (40 and 60%). In greenhouse tests, H. chaetospira effectively controlled clubroot (reducing the disease by 90 to 100%) at pathogen resting spore densities of 104 and 105 spores/g of soil at all soil pHs tested (5.5, 6.3, and 7.2). However, when the resting spore density was 106 spores/g of soil, plants were severely diseased, regardless of treatment, and H. chaetospira had no effect on disease. At a soil moisture content of 40%, disease occurrence was low, regardless of pathogen spore density, but disease was significantly lower in H. chaetospira-treated plants at pathogen spore density of 105 spores/g of soil. At 60% soil moisture content, H. chaetospira significantly could affect at pathogen spore densities of 104 and 105 but not 104/g of soil. At 80% soil moisture content, there was no effect of H. chaetospira at pathogen density. In situ, the soil moisture contents were constantly adjusted to relatively low to moderate (pF 2.2 to 2.4 and pF 2.0 to 2.2) and high (pF 1.6 to 1.8). Other environmental conditions, such as resting spore density and soil pH, were maintained at constant levels. Control plants (not treated with H. chaetospira) showed uniformly high disease levels and proportions of diseased plants across all three moisture treatments (disease index = 72 to 80, proportion of diseased plants 85 to 97%). In the field, H. chaetospira-treated plants at low soil moisture (pF 2.2 to 2.4, plot 1) had 68% disease reduction compared with untreated controls and 49% reduction at moderate moisture pF (pF 2.0 to 2.2, plot 2). There was no effect on disease by H. chaetospira at high soil moisture (pF 1.6 to 1.8, plot 3). Based on our results, H. chaetospira is an effective biocontrol agent against clubroot in Chinese cabbage at a low to moderate soil moisture range and a pathogen resting spore density of 105 (or lower resting spores per gram of soil in situ.

Immune responses against replication-deficient adenovirus inhibit ovalbumin-specific allergic reactions in mice.

Replication-deficient adenovirus vector (Ad) is one of the most efficient gene transfer vehicles for human gene therapy. However, Ad is antigenic, known to evoke prominent inflammatory responses in vivo, and there are concerns that using Ad in patients with immune-mediated disorders (allergy and autoimmune diseases) may affect the status of the diseases. To evaluate this concept in a manner close to clinical scenarios, a mouse model of airway eosinophilic inflammation was developed by administering intraperitoneal injections and inhalations of chicken ovalbumin (OA), with Ad administered intranasally 5 days after the OA sensitization. The administration of Ad resulted in a significant suppression of eosinophil counts in peripheral blood as well as in the bronchoalveolar lavage fluid (BALF), and a decrease in OA-specific IgE. The decrease in the number of eosinophils in BALF was associated with a marked upregulation of interferon gamma (IFN-gamma) expression. In contrast, the Ad-specific, delayed-type hypersensitivity response and efficacy of reporter gene expression mediated by Ad were only marginally affected in animals sensitized with OA. Together, these data support the idea that Ad administration in patients with Th2-mediated immune disorders does not exacerbate the parameters of ongoing inflammations or gene transfer efficiency, and with its ability to induce prominent type 1 immune response to the antigen in vivo, Ad could potentially be used as an efficient adjuvant to control immune disorders where Th2 cell-mediated mechanisms are involved.

Transfer of heme oxygenase 1 cDNA by a replication-deficient adenovirus enhances interleukin 10 production from alveolar macrophages that attenuates lipopolysaccharide-induced acute lung injury in mice.

By using a direct, intratracheal inoculation of an adenovirus encoding heme oxygenase 1 (Ad.HO-1), model gene therapy for acute lung injury induced by inhaled pathogen was performed. Data demonstrated that Ad.HO-1 administration is as effective as the pharmacologic upregulation of the endogenous HO-1 gene expression by hemin to attenuate neutrophilic inflammations of the lung after aerosolized lipopolysaccharide (LPS) exposure. Interestingly, immunohistochemical analysis revealed that the HO-1 gene was transferred not only to the airway epithelium, but to the alveolar macrophages (AMs). Moreover, overexpression of exogenous HO-1 in the macrophages provided a high level of endogenous interleukin 10 (IL-10) production from the macrophages, and additional experiments using IL-10 knockout mice demonstrated that the increase in IL-10 in the macrophages was critical for the resolution of neutrophilic migration in the lung after LPS exposure. These results suggest that AMs not only are barriers for efficient gene transfer to the respiratory epithelium, but also represent logical targets for Ad-mediated, direct, in vivo gene therapy strategies for inflammatory disorders in humans.

Control of Verticillium Yellows in Chinese Cabbage by the Dark Septate Endophytic Fungus LtVB3.

ABSTRACT Three hundred forty-nine fungal endophytes were obtained from a total of 1,214 root segments of eggplant, melon, barley, and Chinese cabbage grown as bait plants in a mixed soil made up of samples from different forest soils in Alberta and British Columbia, Canada. Three of the 349 isolates, when inoculated in axenically reared Chinese cabbage seedlings grown in petri dishes, almost completely suppressed the effects of a postinoculated and virulent strain of Verticillium longisporum. Two isolates effective against the pathogen were Phialocephala fortinii, which had been obtained from the roots of eggplant and Chinese cabbage. The third isolate was a dark septate endophytic (DSE) fungus obtained from barley roots. Hyphae of P. fortinii grew along the surface of the root and formed microsclerotia on or in the epidermal layer. Hyphae of the DSE fungus heavily colonized root cells of the cortex. Seedlings grown for 1 week in the presence of the endophytes were then challenged with the Verticillium pathogen. In DSE-treated roots, some of cell walls in the epidermal and cortical layers showed cell wall appositions and thickenings, which appeared to limit the ingress of the pathogen into adjacent cells. Such marked host reactions were not observed in the root cells colonized by P. fortinii. Chinese cabbage preinoculated with the above endophytes and, for comparison, a previously reported disease-suppressive fungal endophyte, Heteroconium chaetospira, were transplanted into the field and disease symptoms were assessed. The DSE could most effectively inhibit the development of Verticillium yellows, with reductions in the percentages of external and internal disease symptoms of 84 and 88%, respectively. The protective values against the disease are extremely high compared with those of other isolates. Most of the DSE-treated plants in the plots achieved marketable quality.

Retro-odontoid massive calcium pyrophosphate crystal deposition--case report.

An 86-year-old male presented with progressive myelopathy due to retro-odontoid massive deposits of calcium pyrophosphate dihydrate (CPPD) crystals. Magnetic resonance imaging revealed a non-enhanced isointense extradural mass on the T1-weighted image and heterogeneously intense mass on the T2-weighted image. Computed tomography showed typical punctate and linear calcifications within the mass. The mass was resected via a lateral approach resulting in marked improvement of the symptoms. Histological examination revealed birefringent rhomboid crystals consistent with CPPD. CPPD deposition should be considered in the differential diagnosis of retro-odontoid extradural mass because surgical therapy is beneficial even for elderly patients.

[A case of multiple organ failure with massive intestinal bleeding caused by methicillin-resistant Staphylococcus aureus in a postcystectomy patient--efficacy of mask continuous positive airway pressure training and intraarterial embolization].

A 51-year-old man underwent radical cystectomy with tubeless cutaneous ureterostomy. Methicillin-resistant Staphylococcus aureus (MRSA) enteritis developed postoperatively. MRSA caused critical infections such as pneumonia and sepsis, which subsequently progressed to adult respiratory distress syndrome, massive melena and multiple organ failure. The patient was rescued by intensive management including mask continuous positive airway pressure, systemic vancomycin administration and intraarterial embolization to stop jejunal bleeding.

Vaginal stone in a male patient with true hermaphroditism.

A 31-year-old male with true hermaphroditism and a 46, XX karyotype who underwent gonadectomy and extirpation of the internal sex organs at the age of 4 had a large stone 4 cm in diameter in the residual male vagina. He complained of pain on micturition, hematuria, and rectal pressure. Urethroscopy and retrograde urethrography disclosed an ostium of the male vagina in the prostatic urethra, and an impacted intravaginal stone. Transurethral electrohydraulic lithotripsy was performed. Extracorporeal shock wave lithotripsy and transurethral lithotripsy were performed for the residual stones. All stones and fragments were spontaneously passed. The stone was composed of calcium phosphate and ammonium acid urate.

[Unilateral and synchronous occurrence of renal cell carcinoma and ureteral tumor: a case report].

An 81-year-old woman was admitted to our hospital with left flank pain. Excretory urography revealed left hydronephrosis. Abdominal computed tomography (CT) revealed a large heterogenous tumor in the upper pole and marked hydronephrosis and hydroureter in the lower portion of the left kidney. Left total nephroureterectomy was performed under the diagnosis of renal pelvic and ureter tumor. The pathological diagnosis was of renal cell carcinoma (spindle type, grade 3) in the kidney and transitional cell carcinoma (grade 2) in the ureter. Postoperative chemotherapy was not given. Convalescence was uneventful and fifteen months after the operation she is alive with no recurrence or metastasis.

[Treatment of renal cell carcinoma extending into the right atrium with extra-corporeal circulation using high-grade hypothermia: a case report].

A 68-year-old woman underwent surgical treatment for renal cell carcinoma associated with tumor thrombus extending into the right atrium. Although the tumor thrombus reached the level of the right atrium, there were no other apparent metastases. Combination therapy with interferon alfa plus tegafur/uracil (UFT) was attempted with the expectation of reducing the tumor thrombus, but there was no change. Successful management was achieved with right radical nephrectomy, right auriculotomy, and partial cavectomy using cardiopulmonary bypass under high-grade hypothermia. After removal of the tumor and thrombus, blood loss was 13,900 ml during the patient's recovery. She had mild heart failure for about two weeks after the operation, but recovered. She was discharged on the 40th day after the operation. Proper preparation for blood transfusion is the key point of this operation.

[A case report of use of horseshoe kidney as renal transplant from live donor].

This is a report on a surgery performed in February, 1995 describing the donation of a living donor's horseshoe kidney used for renal transplantation. The recipient was a 31 years-old male on hemodialysis since 1994. The donor was the healthy 55 years-old father of the recipient who had an uncomplicated horseshoe kidney. The isthmus was perfused by an accessory artery. Via transperitoneal approach, the horseshoe kidney was mobilized for in situ perfusion. A microwave coagulator was used to divide the isthmus, and the cut surfaces were closed by mattress sutures and fibrin glue. The left kidney was transplanted into the recipient's right iliacfossa. While his post-transplant course was complicated by urinary leakage, the graft remained free of rejection until and beyond the 6 months post transplant period when he was discharged at s-Cr 1.7 mg/dl. The donor's convalescence was uneventful. During the 20 months post-transplant period both the donor and recipient are doing well.

[The efficacy of re-TUR after 3 weeks of initial TUR treatment for locally advanced bladder cancer].

BACKGROUND: Radical cystectomy was usually performed for the patient with advanced bladder cancer. The choice of surgical procedure whether bladder preserved therapy or radical cystectomy has sometime plagued us in bladder cancer treatment. Because we don't have clear guideline for the treatment of locally advanced bladder cancer (T2-T3a N0M0). We devised a radical treatment for the patients with locally advanced bladder cancer without radical cystectomy. PATIENTS AND METHODS: We performed re-TUR treatment after 3 weeks of initial TUR for 13 patients with grade 2 and locally advanced bladder cancer diagnosed with pelvic CT scan, transurethral ultrasonography or bimanual examination under the anesthesia. RESULTS: We could successfully preserve all cases of bladder with the use of this devised TUR (periods 9-45 months median 22 months). This method has significantly reduced the cystectomy rate in such locally advanced cases compared with the cystectomy rate prior to 1993 (p < 0.001). CONCLUSION: This method has significance on treatment for locally advanced papillary bladder cancer as bladder preserving therapy with careful observation.

Minimal incision transinguinal repair for incarcerated obturator hernia.

BACKGROUND: Patients with incarcerated obturator hernia are usually elderly, frail, and physically inactive women with serious comorbidities. Although a laparotomy is standard surgical intervention for emergency incarcerated or strangulated obturator hernia, it is invasive particularly for these high-risk patients. The aim of this study is to show the feasibility of minimum open inguinal approach to reduce surgical risk for preoperatively diagnosed incarcerated obturator hernia. METHODS: Between April 2008 and July 2012, 3 consecutive incarcerated obturator hernia patients at Kamitsuga General Hospital who were diagnosed preoperatively by computed tomography underwent the following procedure. First a 4 cm inguinal hernia incision and preperitoneal dissection through the opening of the deep inguinal ring are made. The obturator hernia can be easily found 2 cm dorsally from the Cooper's ligament extraperitoneally. A small incision is made at medial sharp edge of the hernia defect. The hernia sac and its content can then be reduced. If the incarcerated bowel is viable, a prosthetic mesh is placed as a patch. If the bowel is necrotic, the damaged bowel loop is withdrawn through the wound and easily reconstructed extra-abdominally. RESULTS: All operations were successfully completed with this procedure. All patients recovered without incident. CONCLUSIONS: Minimal incision transinguinal repair for diagnosed incarcerated obturator hernia is feasible and provides an improved option to more invasive procedures.

Unique DNA plasmid pRS64 associated with chromosomal DNAs of the plant pathogenic fungus Rhizoctonia solani.

Unique DNA sequences homologous to the linear DNA plasmid pRS64 were investigated in chromosomal DNAs of isolates belonging to anastomosis group 4 (AG-4) of the plant pathogenic fungus Rhizoctonia solani. Chromosome-sized DNAs of isolates RI-64 and 1271 of AG-4 were separated into six bands by orthogonal-field-alternation gel electrophoresis and hybridized to a cloned segment of pRS64. A small chromosome-sized DNA band of approximately 1.1 Mb carried the sequences homologous to pRS64 DNA. Sequences homologous to pRS64 were also maintained within the chromosomal DNA of isolate 127.1 of AG-4 which does not possess the plasmid. The plasmid showed no homology to the mitochondrial DNA of isolate 1271. The possibility that the linear plasmid pRS64 may act as a transposable genetic element is discussed.

Role of the gynoecium in natural senescence of carnation (Dianthus caryophyllus L.) flowers.

Although the role of the gynoecium in natural senescence of the carnation flower has long been suggested, it has remained a matter of dispute because petal senescence in the cut carnation flower was not delayed by the removal of gynoecium. In this study, the gynoecium was snapped off by hand, in contrast to previous investigations where removal was achieved by forceps or scissors. The removal of the gynoecium by hand prevented the onset of ethylene production and prolonged the vase life of the flower, demonstrating a decisive role of the gynoecium in controlling natural senescence of the carnation flower. Abscisic acid (ABA) and indole-3-acetic acid (IAA), which induced ethylene production and accelerated petal senescence in carnation flowers, did not stimulate ethylene production in the flowers with gynoecia removed (-Gyn flowers). Application of 1-aminocyclopropane-1-carboxylate (ACC), the ethylene precursor, induced substantial ethylene production and petal wilting in the flowers with gynoecia left intact, but was less effective at stimulating ethylene production in the -Gyn flowers and negligible petal in-rolling was observed. Exogenous ethylene induced autocatalytic production of the gas and petal wilting in the -Gyn flowers. These results indicated that ethylene generated in the gynoecium triggers the onset of ethylene production in the petals of carnation during natural senescence.

Expression of the linear DNA plasmid pRS64 in the plant pathogenic fungus Rhizoctonia solani.

The plant pathogenic isolate RI-64 of anastomosis group 4 of Rhizoctonia solani possesses three linear DNA plasmids (pRS64-1, -2, and -3). Unique poly(A)- RNA, 0.5 kb in length and hybridizable with the pRS64 DNAs was found in mycelial cells of the isolate RI-64. The overall homology at the nucleotide level between pRS64-1, -2, and -3, and the cDNA prepared from the poly(A)- RNA was 100%, 73%, and 84%, respectively. The open reading frames found in pRS64-1, -2, and -3 (ORF1-1, ORF2-1, and ORF3-1) are 68 amino acids long. The amino acids sequence showed no significant homology with known proteins. Extracts from Escherichia coli cells expressing ORF1-1 contain a specific protein of 7 kDa. Antisera raised against the ORF1-1 product obtained from E. coli cells cross-reacted with the specific proteins found in the mycelia. The results indicate that the DNA plasmids found in R. solani contain a sequence that encodes a specific protein which may be involved in determination of plant pathogenicity.

[High pressure liquid chromatography of corticosteroids: I. Preliminary report of the determinations of plasma corticosterone by HPLC combined with radioimmunoassay (author's transl)].

The analysis of corticosteroids (CDS) was preliminarily investigated using a DU PONT 830 High Pressure Liquid Chromatography (HPLC) equipped with a UV detector and a Permaphase column ETH. 1) Firstly, the best condition for the separation of mixed CS was investigated. In a linear gradient elution, ten authentic steroids, varying greatly in polarity (including native CS such as DOC, Corticosterone and Cortisol) were sharply separated in about 20 minutes. Recovery of the steroid injected into the column was 96 approximately 100%. 2) Standard CS added to the chloroform extracts from plasma or urine were also clearly separated in the same condition as above. In the experiment using predonisolone and estrone-propionate in different doses, the synthetic steroids were quantitatively separated, giving a linear calibration curve. 3) Sensitivity of the steroid determination was ng order photochemically in a plot study, but the method, using a UV detector, was not sufficient to apply to small amount of samples extracted with choloroform. To solve this problem, radioimmunoassay was applied to the eluate containing biological steroid, which was roughly separated using the synthetic steroid marker. As for the determination of plasma corticosterone using this method of HPLC combined with radioimmunoassay, the results were satisfactory for practical use; the mean based values and SD of 10 normal controls were 0.4+/-0.2 microgram/100ml for corticosterone, and they showed 4 to 5-fold increases to 250 microgram of synthetic ACTH injection.