Increased metabolite production by deletion of an HDA1-type histone deacetylase in the phytopathogenic fungi, Magnaporthe oryzae (Pyricularia oryzae) and Fusarium asiaticum.

Histone deacetylases (HDACs) play an important role in the regulation of chromatin structure and gene expression. We found that dark pigmentation of Magnaporthe oryzae (anamorph Pyricularia oryzae) ΔMohda1, a mutant strain in which an orthologue of the yeast HDA1 was disrupted by double cross-over homologous recombination, was significantly stimulated in liquid culture. Analysis of metabolites in a ΔMohda1 mutant culture revealed that the accumulation of shunt products of the 1,8-dihydroxynaphthalene melanin and ergosterol pathways were significantly enhanced compared to the wild-type strain. Northern blot analysis of the ΔMohda1 mutant revealed transcriptional activation of three melanin genes that are dispersed throughout the genome of M. oryzae. The effect of deletion of the yeast HDA1 orthologue was also observed in Fusarium asiaticum from the Fusarium graminearum species complex; the HDF2 deletion mutant produced increased levels of nivalenol-type trichothecenes. These results suggest that histone modification via HDA1-type HDAC regulates the production of natural products in filamentous fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural products of fungi have significant impacts on human welfare, in both detrimental and beneficial ways. Although HDA1-type histone deacetylase is not essential for vegetative growth, deletion of the gene affects the expression of clustered secondary metabolite genes in some fungi. Here, we report that such phenomena are also observed in physically unlinked genes required for melanin biosynthesis in the rice blast fungus. In addition, production of Fusarium trichothecenes, previously reported to be unaffected by HDA1 deletion, was significantly upregulated in another Fusarium species. Thus, the HDA1-inactivation strategy may be regarded as a general approach for overproduction and/or discovery of fungal metabolites.

Multiple comparison among groups of growth curves.

The problem of comparing a sequence of independent experiments divided into several groups with a control is discussed under the logistic growth-curve models. We propose a method for constructing multiple testing procedures using the closed testing procedures and the random-effect model for summarizing estimated values of parameters.

Point process models in asthma attacks for assessing environmental risk factors.

Point process models are reviewed and discussed for assessing the effects of environmental risk factors on asthma attacks. It is pointed out that the logit model and proportional intensity model are useful for analyzing the data based on the diaries recorded consecutively during several months or during a few years. Some covariates that seems to influence upon asthmatics are explored using these models. Further work on estimating the smoothed base-line intensity function is briefly discussed in terms of the Bayes model.

Neural invasion in gastric carcinoma.

AIMS: To determine whether neural invasion in advanced gastric cancer is of clinicopathological significance. METHODS: The study population comprised 121 cases of primary advanced gastric carcinoma. Two paraffin wax embedded blocks taken from the central tissue slice in each primary tumour were used. For definitive recognition of neural invasion, immunostaining for S-100 protein was applied to one slide; the other slide was stained with haematoxylin and eosin. RESULTS: Neural invasion was recognised in 34 of 121 (28%) primary gastric carcinomas. There were significant differences in tumour size, depth of tumour invasion, stage, and curability between patients with and without neural invasion. The five year survival rates of patients with and without neural invasion were 10 and 50%, respectively. Multivariate analysis, however, demonstrated that neural invasion was not an independent prognostic factor. CONCLUSIONS: Neural invasion could be an additional useful factor for providing information about the malignant potential of gastric carcinoma. This may be analogous to vessel permeation which is thought to be important, but is not an independent prognostic factor.

Chromogranin positive cells in colorectal carcinoma and transitional mucosa.

AIMS: Immunostaining of chromogranin identifies gastrointestinal mucosal endocrine cells. The detailed distribution and significance of chromogranin positive cells in colorectal carcinomas and in transitional mucosa remain unclear. The aim of this study was to clarify these aspects. METHODS: The distribution of chromogranin positive cells was studied by immunohistochemical methods in normal epithelium remote from carcinoma, in transitional mucosa, and in carcinomas of the colorectum. In selected cases northern or western blot analyses were performed. RESULTS: Chromogranin positive cells were seen in the lower third of the normal crypts and less frequently in transitional mucosa. Thirty five per cent (n = 38) of colorectal carcinomas showed immunohistochemically positive carcinoma cells in the tumour tissue. Northern and western blot analyses showed similar results. There was no difference in clinicopathological factors, including prognosis, between chromogranin positive cases of colorectal carcinoma (n = 38) and chromogranin negative cases (n = 70). CONCLUSIONS: Neuroendocrine cell differentiation is controlled in transitional mucosa and the presence of chromogranin positive cells in carcinoma tissue does not influence the patient's prognosis.

Role of lymph node dissection and splenectomy in node-positive gastric carcinoma.

BACKGROUND: In gastric carcinoma the relationship between survival of patients and degree of lymph node dissection is still controversial and there is no agreement about the efficacy of extended lymph node dissection and splenectomy. METHODS: Long-term results after surgery were statistically analyzed in 240 patients who underwent curative gastrectomy with lymph node dissection for node-positive gastric carcinoma from 1975 to 1990. RESULTS: The survival of patients was not significantly different among R1, R2, and R3 dissections, with the 10-year survival rate being 57%, 50%, and 44%, respectively. The survival rate was also not different between R2 and R3, even when the cases were stratified by the level of lymph node metastasis (N1, 66% vs 58%; N2, 28% vs 31%). The survival of patients was not significantly different between cases treated without splenectomy and those treated with splenectomy, with 10-year survival rates being 47% and 26%, respectively. The survival rate was also not different between non-splenectomy- and splenectomy-treated cases, even when the patients were compared in the same level of lymph node metastasis (N1, 60% vs 45%; N2, 23% vs 11%). CONCLUSIONS: In cases of curative gastrectomy the survival time of patients was not influenced by the level of lymph node dissection and splenectomy. For the treatment of node-positive gastric carcinoma (N1, N2) the long-term efficacy of R3 dissection and splenectomy seems limited.

Effect of nafamostat mesilate on coagulation and fibrinolysis in hepatic resection.

The effect of nafamostat mesilate on coagulation and fibrinolysis was investigated in a study of 22 patients with hepatocellular carcinoma who underwent a hepatic resection. The patients were divided into two groups: group 1, control (n = 11) and group 2, those with the intraoperative and postoperative use of nafamostat mesilate (0.2 to 0.4 milligram per kilogram per hour, n = 11). Nafamostat mesilate tended to suppress the coagulation expressed by thrombin-antithrombin III complex and fibrinopeptide A both during and immediately after operation. Moreover, nafamostat mesilate significantly suppressed the fibrinolysis expressed by euglobulin lysis activity both during and after operation. With regard to the initial stage of the fibrinolytic system, such as tissue-type plasminogen activator and plasminogen activator inhibitor-1, there was no difference between the groups. Therefore, the suppression of the euglobulin lysis activity may be caused by the inhibition of plasmin activity. There was no difference between the groups regarding operative blood loss. However, the rate of blood transfusion in group 2 was lower than that in group 1, and no fresh frozen plasma was required for the patients who lost over 2,000 milliliters of blood. Nafamostat mesilate can suppress euglobulin lysis activity both intraoperatively and postoperatively, and thus decrease the amount of blood transfusion needed. Therefore, at present, nafamostat mesilate seems to be one of the most useful agents for stabilizing the coagulant and fibrinolytic systems in hepatic resection.

Changes in regulating blood coagulation in hepatic resection with special references to soluble thrombomodulin and protein C.

The protein C anticoagulant pathway in hepatic resection was studied. The patients were divided into two groups--group 1 consisted of patients with a normal liver and group 2 consisted of patients with either hepatitic or a cirrhotic liver. Plasma protein C activity and soluble thrombomodulin were then sequentially measured during hepatectomy and in the early postoperative period. The protein C activity in group 1 decreased during hepatectomy and reached a low immediately after operation, and thereafter, recovered to near preoperative levels. However, the preoperative value in group 2 was lower than that in group 1 and the postoperative values were significantly lower than those in group 1 (p < 0.05). The level of soluble thrombomodulin in group 1 decreased during hepatectomy but later returned to preoperative levels. However, in group 2, the preoperative value was higher than that in group 1 and the postoperative values were greater than that of the preoperative values, while the values were significantly higher than those in group 1 (p < 0.05). During hepatectomy, hypercoagulability may contribute to the low levels of protein C and soluble thrombomodulin. The postoperative significant increase of soluble thrombomodulin may, thus, indicate the occurrence of endothelial injury in the remnant liver. The sequential measurements of both parameters can, therefore, be useful in detecting coagulopathy and endothelial injury in hepatic resection.

Modulation of coagulation and fibrinolysis in hepatic resection: a randomized prospective control study using antithrombin III concentrates.

A randomized prospective control trial for determining the efficacy of antithrombin III concentrates in hepatic resection was performed using 24 patients with hepatocellular carcinoma. Thirteen patients were given antithrombin III concentrates (1,500 IU) immediately before operation, during hepatectomy and immediately after operation. Coagulant and fibrinolytic profiles were determined by molecular markers such as thrombin-antithrombin III complex and plasmin-alpha 2plasmin inhibitor complex. During hepatic resection, both hypercoagulability and mainly primary hyperfibrinolysis occurred. Regarding the effectiveness of antithrombin III concentrates, in the antithrombin III treatment group, only a significant lower incidence of positive soluble fibrin monomer complex at postoperative days 1 and 5 was found among all the parameters studied. Therefore, no definite evidence of clinical usefulness of the perioperative administration of antithrombin III concentrates in hepatic resection was proved.

Immunohistochemical expression of sialyl Tn and sialyl Lewis(a) antigens in stromal tissue correlates with peritoneal dissemination in stage IV human gastric cancer.

Immunohistochemical expression of sialyl Tn antigen (TKH2 and B72.3), sialyl Lewis(a) antigen (CA19-9) and CEA was examined in 52 cases with Stage IV gastric cancer, the objective being to examine whether or not these tumour-associated antigens show different expression between peritoneal dissemination and liver metastasis. Stromal type for TKH2, B72.3, CA19-9 and CEA were seen in 14 (50%), 13 (46%), 18 (64%) and nine (32%) cases with peritoneal dissemination, respectively, and in four (17%), three (13%), four (17%) and four (17%) cases with liver metastasis, respectively. The stromal type correlated to peritoneal dissemination in TKH2 (P < 0.05) B72.3 (P < 0.05) and CA19-9 (P < 0.05). In other clinicopathological factors, the stromal type for TKH2, B72.3 and CA19-9 correlated to the undifferentiated type. However, even in the undifferentiated type group, the stromal type correlated to peritoneal dissemination in TKH2 (P < 0.05), B72.3 (P < 0.05) and CA19-9 (P < 0.05). Based on these results, the spread of sialyl-rich tumour-associated antigens into the surrounding stroma in the undifferentiated type of gastric cancer may be associated with peritoneal dissemination.

A simple classification of lymph node level in gastric carcinoma.

BACKGROUND: Because insufficient lymph node examination can cause erroneous listing of earlier-nodal-stage gastric carcinoma (stage migration phenomenon), surgical results must be evaluated based on a highly accurate examination of the dissected lymph nodes. To establish a simple and useful classification of lymph node level, we analyzed the frequency and distribution of lymph node metastasis by using curatively treated node-positive gastric carcinoma. PATIENTS AND METHODS: Various clinicopathologic data were analyzed with reference to the degree of lymph node metastasis by using 240 patients with curatively resected node-positive gastric carcinoma. The cases were divided into the following three groups: 142 with positive level I (perigastric) nodes, 71 with positive level II (intermediate) nodes, and 27 with positive level III (distant) nodes, irrespective of the location of tumors. RESULTS: The level of lymph node metastasis clearly correlated with the survival of patients, with the 5-year survival rates for level I, II, and III cases being 67%, 35%, and 26%, respectively (P < 0.01). The degree of lymph node metastasis was determined by the number of positive nodes (P < 0.01), the depth of invasion (P < 0.01), the size of tumors (P < 0.01), and the location of tumors (P < 0.05). CONCLUSION: This simple classification of lymph node level (level I, II, and III) is useful for the evaluation and prediction of surgical results in gastric carcinoma.

Estimation of risk of major complications after hepatic resection.

To identify the risk factors predicting major postoperative complications from among preoperative and intraoperative variables, an extensive retrospective analysis of 209 patients who underwent hepatic resections was performed using multivariate logistic regression. The major complications were defined as liver failure, intractable ascites and pleural effusion, intraperitoneal infection, intra-abdominal hemorrhage requiring reoperation, major bile leakage, and gastrointestinal tract bleeding. First, detailed pre- and intraoperative data including medical history, laboratory data, portion and extent of hepatectomy, operative time, and amount of blood loss were univariately analyzed. Next, any significant variables were multivariately analyzed using the logistic regression method. Diabetes, increased intraoperative blood loss, resection of segment 8, and an increased serum blood urea nitrogen level were independent and significant variables predicting major postoperative complications. A higher level of serum cholesterol and a procedure involving a portion of left lateral segment were found to decrease the risk. Both more careful operative procedures and intensive management of DM and renal dysfunction in the perioperative period could result in a better quality of life after hepatic resection.

Patient registration and treatment allocation in multicenter clinical trials using a FAX-OCR system.

This article describes the design and results of implementation of an automated patient registration and treatment allocation system (RETAS) used in multicenter clinical trials. RETAS was developed using a FAX-OCR system by which handwritten Japanese and English characters, as well as numericals and forms with check boxes, are sent from participating institutions by Fax, processed using an optical character reader, and then transmitted to a host computer at a statistical center. Based on the facsimile data, RETAS can automatically review eligibility, collect patient identification data and provide a randomized treatment allocation. RETAS permits uninterrupted, unattended operation at a statistical center, 24 hours a day, 7 days a week. Therefore, it drastically decreases the workload of personnel at the statistical center needed to support central telephone registration coverage. Consequently, staff members are free to focus on patient registration, treatment allocation, and follow-up of patients. The treatment allocation procedure in this system is based on Pocock and Simon's minimization method combined with Zelen's method for institution balancing. By this system it was possible to balance treatment numbers for each level of various prognostic factors over an entire trial and, at the same time, balance the allocation of treatments within an institution. The system currently supports the protocol of a clinical trial for Adjuvant Chemo-Endocrine Therapy for Breast Cancer in West Japan.

Significance of intraoperative measurement of liver consistency prior to hepatic resection.

The functional reserve of the liver should be estimated prior to hepatectomy. While the experienced liver surgeon often relies on the findings of manual palpation intra-operatively, this approach remains a subjective one. In 22 of our patients scheduled for hepatectomy, we measured intra-operatively hepatic consistency as well as the parameters of liver function. Hepatic consistency is augmented during progression from non-fibrosis to cirrhosis. We found a statistically significant correlation between liver consistency and the indocyanine green retention rate at 15 minutes (r = 0.682; p = 0.0009) and portal vein pressure (r = 0.733; p = 0.0008). Prothrombin time, the hepaplastin test, total bilirubin and glutamic pyruvic transaminase showed no significant correlation with hepatic consistency. The safety limits of the hepatic blood circulation for the performance of hepatectomy can be estimated using parameters of consistency in this organ.

Indicative value of carcinoembryonic antigen (CEA) for liver recurrence following curative resection of stage II and III gastric cancer.

BACKGROUND/AIMS: We investigated the role of preoperative and serial postoperative serum CEA levels in advanced gastric cancer patients with curative operation. MATERIALS AND METHODS: Preoperative and serum postoperative CEA levels were measured in 115 patients with Stage II and III gastric cancer and who underwent curative gastric resection. RESULTS: The 5-year survival rates of patients with high preoperative CEA levels (> 5 ng/ml) were poorer than in case of low preoperative CEA levels (< 5 ng/ml) in both stage II (P < 0.05) and stage III (P < 0.01) gastric cancer. When the site of the recurrence was diagnosed in 47 patients with less than a 5-year survival, a high level of preoperative CEA was more likely to be associated with a liver metastasis (13/18, 72.2%) than with peritoneal dissemination (4/15, 26.7%) (P < 0.05). In the course of serial measurements of postoperative levels of CEA, CEA increased in patients with liver metastasis, and CEA levels began to elevate 3.2 months before clinical detection, while, little change of postoperative serum CEA levels was noted in patients with peritoneal dissemination. CONCLUSIONS: We propose that the preoperative and serial postoperative assays of serum CEA level are predictive for liver metastasis in patients with stage II and III gastric cancer and treated by curative gastric resection.

Influence of blood transfusion on postoperative long-term liver function in patients with hepatocellular carcinoma.

BACKGROUND/AIMS: To research for possible adverse effects of blood transfusion on liver function at a relatively long term after hepatic resection, liver function tests at one and two years after the surgery were studied in patients with hepatocellular carcinoma. MATERIALS AND METHODS: Seventy-one patients with no evidence of cancer recurrence during two years after the surgery were selected for this retrospective analysis. Patients were divided into two groups depending on perioperative blood transfusion: those who received blood transfusion (BT(+), n = 38), and those without blood transfusion (BT(-), n = 33). RESULTS: There were no statistical difference in the preoperative clinical features including liver function tests except decreased prothrombin time index (P < 0.01) and larger amount of intraoperative blood loss (P < 0.01) in BT(+) group. Platelet counts at 2 year after the surgery were significantly higher in the BT(-) group (P < 0.05). Two-way ANOVA showed no significant difference in liver function tests between the two groups. In multiple regression analysis, blood transfusion significantly correlated with serum total bilirubin at 1 year, but did not at 2 years after the surgery. CONCLUSIONS: The influence of blood transfusion in the patients with hepatocellular carcinoma was considered to be minimum within 2 years after hepatectomy.

Model equation for strongly focused finite-amplitude sound beams

A model equation that describes the propagation of sound beams in a fluid is developed using the oblate spheroidal coordinate system. This spheroidal beam equation (SBE) is a parabolic equation and has a specific application to a theoretical prediction on focused, high-frequency beams from a circular aperture. The aperture angle does not have to be small. The theoretical background is basically along the same analytical lines as the composite method (CM) reported previously [B. Ystad and J. Berntsen, Acustica 82, 698-706 (1996)]. Numerical examples are displayed for the amplitudes of sound pressure along and across the beam axis when sinusoidal waves are radiated from the source with uniform amplitude distribution. The primitive approach to linear field analysis is readily extended to the case where harmonic generation in finite-amplitude sound beams becomes significant due to the inherent nonlinearity of the medium. The theory provides the propagation and beam pattern profiles that differ from the CM solution for each harmonic component.

The value of prophylactic R2 lymph node dissection for gastric cancer.

Three hundred and eleven patients with an R2 lymph node dissection for gastric cancer were reviewed in order to examine the efficacy of prophylactic R2 lymph node dissection. In 246 patients, who were all macroscopically determined to have either N0 or N1 lymph node metastasis at the time of operation, 30 showed histologic lymph node metastasis in the N2 group. When the 5-year survival rates in the 61 patients with histologic N2 lymph node metastasis were compared according to the macroscopic evaluation of lymph node metastasis at the time of operation, 30 patients who were determined to have either N0 or N1 lymph node metastasis showed a better prognosis than the 31 patients who were determined to have N2 lymph node metastasis (p < 0.01). Therefore, prophylactic R2 lymph node dissection contributes to an improved prognosis for gastric cancer patients who are found to macroscopically have either N0 or N1 lymph node metastasis on the basis of an intra-operative evaluation.

[Usefulness of epidural infusion of ketamine for relief of localized superficial pain].

Three patients with localized superficial pain had their pain alleviated by single epidural infusion with low dose ketamine. The patients are as follows: a 62-year-old female with herpetic neuralgia on her right sixth thoracic nerve area; a 52-year-old male whose left shoulder, anterior chest and abdomen had been burned by acetylene gas; and a 49-year-old male whose bilateral hands suffering from frostbite by propane gas. Epidural tube insertion to administer a single dose of 10 mg ketamine with lidocaine or bupibacaine was performed in all the three patients. They were administered single epidural infusion of 10 mg ketamine with lidocaine or bupibacaine everyday and they continued to receive epidural block with lidocaine or bupivacaine including buprenorphine or morphine. Therefore, we suspect that single epidural infusion of ketamine, an antagonist for N-methyl-D-aspartic acid receptor, could be an effective and useful alternative treatment in patients with various refractory localized superficial pain of either acute or chronic nature.

[Treatment with stellate ganglion block, continuous epidural block and ulnar nerve block of a patient with postherpetic neuralgia who developed complex regional pain syndrome (CRPS)].

We present a case of a 46-year-old female patient with systemic lupus erythematosus who developed herpes zoster of the right eighth cervical nerve. Her whole right forearm, hand and the first through fifth fingers were coated with some gel and protected against pain. She had been suffering from continuous and spasmodic burning pain, hyperalgesia, allodynia, drop in skin temperature, sudmotor disturbance, edema, constructure of the joints, muscle atrophy and bone atrophy of her right upper extremity probably due to postherpetic neuralgia (PHN) associated with complex regional pain syndrome (CRPS). She received right stellate ganglion block (SGB), continuous cervical epidural block and right ulnar nerve block. Reduction of pain and edema as well as improvement in mobility of each joint of her right upper extremity was observed. We suspect that SGB, continuous cervical epidural block and ulnar nerve block are effective and useful alternative treatments in a patient with PHN associated with CRPS of the eighth cervical nerve.