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Acute gastroenteritis associated with diarrhea is considered a serious disease in Africa and South Asia. In this study, we examined the trends in the causative pathogens of diarrhea and the corresponding gut microbiota in Ghana using microbiome analysis performed on diarrheic stools via 16S rRNA sequencing. In total, 80 patients with diarrhea and 34 healthy adults as controls, from 2017 to 2018, were enrolled in the study. Among the patients with diarrhea, 39 were norovirus-positive and 18 were rotavirus-positive. The analysis of species richness (Chao1) was lower in patients with diarrhea than that in controls. Beta-diversity analysis revealed significant differences between the two groups. Several diarrhea-related pathogens (e.g., Escherichia-Shigella, Klebsiella and Campylobacter) were detected in patients with diarrhea. Furthermore, co-infection with these pathogens and enteroviruses (e.g., norovirus and rotavirus) was observed in several cases. Levels of both Erysipelotrichaceae and Staphylococcaceae family markedly differed between norovirus-positive and -negative diarrheic stools, and the 10 predicted metabolic pathways, including the carbohydrate metabolism pathway, showed significant differences between rotavirus-positive patients with diarrhea and controls. This comparative study of diarrheal pathogens in Ghana revealed specific trends in the gut microbiota signature associated with diarrhea and that pathogen-dependent dysbiosis occurred in viral gastroenteritis.

HIV-1 infected individuals under antiretroviral therapy can control viremia but often develop non-AIDS diseases such as cardiovascular and metabolic disorders. Gut microbiome dysbiosis has been indicated to be associated with progression of these diseases. Analyses of gut/fecal microbiome in individual regions are important for our understanding of pathogenesis in HIV-1 infections. However, data on gut/fecal microbiome has not yet been accumulated in West Africa. In the present study, we examined fecal microbiome compositions in HIV-1 infected adults in Ghana, where approximately two-thirds of infected adults are females. In a cross-sectional case-control study, age- and gender-matched HIV-1 infected adults (HIV+; n = 55) and seronegative controls (HIV-; n = 55) were enrolled. Alpha diversity of fecal microbiome in HIV+ was significantly reduced compared to HIV- and associated with CD4 counts. HIV+ showed reduction in varieties of bacteria including Faecalibacterium, the most abundant in seronegative controls, but enrichment of Proteobacteria. Ghanaian HIV+ exhibited enrichment of Dorea and Blautia; bacteria groups whose depletion has been reported in HIV-1 infected individuals in several other cohorts. Furthermore, HIV+ in our cohort exhibited a depletion of Prevotella, a genus whose enrichment has recently been shown in men having sex with men (MSM) regardless of HIV-1 status. The present study revealed the characteristics of dysbiotic fecal microbiome in HIV-1 infected adults in Ghana, a representative of West African populations.

Aim: As techniques in laparoscopic cholecystectomy (LC) have improved, the role of routine prophylactic abdominal drainage may be limited. A retrospective review was carried out of patients undergoing elective LC to evaluate the benefit of routine drainage in simple uncomplicated procedures. Methods: This study of 295 patients with cholecystolithiasis or gallbladder polyp included 145 patients who underwent LC with drainage and 150 patients who underwent LC without drainage between 2003 and 2007. Allocation to drain or not to drain was non-randomized and based on surgeon preference according to intraoperative findings. Patient characteristics, operative results, and postoperative outcomes were compared between the two groups with univariate analysis. Results: Time to first flatus and length of postoperative hospital stay in the LC without drainage group were shorter than in the LC with drainage group. There was no significant difference between the two groups with respect to postoperative complication rate. No complications were noted due to the lack of drain placement. Conclusion: The use of drain after simple elective uncomplicated LC could safely be limited to appropriate patients as judged by the operating surgeon.

Disruption of the blood-brain barrier (BBB) and/or the blood-cerebrospinal fluid barrier (BCSFB) is thought to be one of the major pathophysiological consequences of meningitis and contributes to the development of adverse neurological outcomes. In order to clarify this hypothesis further, we sequentially quantified the permeability of these barriers with magnetic resonance imaging (MRI) contrast enhancement using gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) in rats with various experimentally-induced meningitis. Meningeal inflammation was elicited by an intracisternal injection of interleukin (IL)-1beta, prostaglandin (PG) E(2), or lipopolysaccharide (LPS). Barrier permeability was calculated from the gadolinium-enhancement ratio (GER) in the subarachnoid space (SAS). The secretion of Gd-DTPA into the SAS was monitored by T1-weighted imaging after an intravenous injection of Gd-DTPA. As a significant linear correlation was observed between the GER and the standard solution, the concentration of the secreted Gd-DTPA were determined from the GER. The maximal intensity in SAS was detected at 5min after Gd-DTPA administration and it declined gradually. Among the inflammatory agents, IL-1beta was found to induce the most severe meningitis as determined from the GER. The concentration of Gd-DTPA in the SAS increased in a dose-dependent manner following IL-1beta intracisternal injection. On the other hand, no significant changes in signal intensity of the brain parenchymal areas due to IL-1beta injection were observed. The findings suggest that the permeability of the BCSFB can be evaluated quantitatively by calculating the GER. MRI with Gd-DTPA provides a useful method to monitor the change in the permeability to the brain barriers.

793 Background: We have reported the effectiveness of neoadjuvant radiotherapy (RT) combined with S-1 in terms of compliance and pathologic response for locally advanced rectal cancer. In the present study, we aimed to evaluate the safety and efficacy of laparoscopic surgery for locally advanced rectal cancer following neoadjuvant RT combined with S-1 in comparison with open surgery. Methods: Two multicenter prospective phase II trials were conducted (UMIN003396, UMIN003398). A total of 63 patients with locally advanced rectal cancer classified as T3-4, N0-3, and M0 were enrolled, and neoadjuvant RT combined with S-1 and total mesorectal excision with D3 lymphadenectomy was performed. Of these, 57 patients were analyzed and divided into a laparoscopic group (LAP, n = 43) and open group (OP, n = 14). We evaluated the short- and long-term outcomes of laparoscopic surgery compared with open surgery by univariate and multivariate analyses. Results: In the patient background, there were no significant differences between the two groups except that cases with T3 and N0 were significantly higher in the LAP compared with the OP. In the operative findings, operation time was longer (mean 447 min vs. 352 min, p = 0.007) and blood loss was lesser (median 220 ml vs. 485 ml, p = 0.033) in the LAP than those in the OP. Although there were no significant differences observed in the incidence of perioperative and late complications between the two groups, reoperation within 30 days was significantly less in the LAP compared with the OP (1 case vs. 5 cases, p = 0.0004). In the multivariate analysis, a distance of the tumor from anal verge within 3 cm was the independent risk factor for reoperation within 30 days. Furthermore, estimated 5-year disease-free survival (LAP 72 % vs. OP 73 %, p = 0.945) and 5-year overall survival (LAP 76 % vs. 75 %, p = 0.836) didn’t significantly differ between the two groups in the Kaplan–Meier curve. Conclusions: The findings of this study demonstrated that laparoscopic surgery for locally advanced rectal cancer following neoadjuvant RT combined with S-1 could be an optional procedure in terms of short- and long-term outcomes. Clinical trial information: UMIN000003396, UMIN000003398.

A case of retroperitoneal mucinous cystadenoma treated laparoscopically is reported. A 26-year-old woman, who complained of abdominal fullness, was referred to us after a large intra-abdominal mass was discovered at her local hospital. Ultrasound and computed tomography showed a fluid-filled mass, 9 cm in diameter, in the left retroperitoneal space. Laparoscopic surgery was performed. We found a smooth and thin-walled cystic tumor that displaced the descending colon to the right and arose from the retroperitoneum, loosely adhering to the psoas muscle. We successfully extirpated the tumor laparoscopically. The surgical specimen consisted of a multilocular, red to yellowish-gray, cystic mass with a smooth, glistening external surface, and inner lining. The mass was filled with thin, yellowish fluid. The histologic diagnosis was benign mucinous cystadenoma. The postoperative course was uneventful, and the patient remains free of recurrence 15 months after surgery. Retroperitoneal mucinous cystadenoma is a rare entity, and the treatment of choice is surgical excision. Laparoscopic extirpation should be considered for the treatment of selected retroperitoneal cystic lesions.

Serologic screening for antibodies to the acquired immunodeficiency syndrome (AIDS) virus human immunodeficiency virus (HIV) has been carried out in Ghana since 1986 when the 1st AIDS case was detected. In general surveys have indicated a low HIV-1 seroprevalence rate among the general population. Of 1852 blood donors screened in 1986 only 2 (0.1%) were positive. None of 168 laboratory workers screened at the same time was seropositive. Also demonstrated has been a relatively low (2.1%) seroprevalence rate among local prostitutes in Accra. However of 151 Ghanaian women who worked as prostitutes in the Ivory Coast 49% were HIV-1 seropositive upon their return to Ghana. Similarly 215 males and females from the general population who had spent time living in the Ivory Coast and other West African countries such as Burkina Faso were found to be seropositive. In all 212 Ghanians have been tested as seropositive for HIV infection in the March 1986-July 1987 period. The greatest number of cases have been reported from Ghanas Eastern Region Greater Accra and Ashanti. The male to female ratio of HIV-positive cases was 1 to 11.6 in 1986 and 1 to 6.3 in 1987. The predominant presenting complaints of AIDS patients in Ghana are weight loss (89%) persistent or intermittent diarrhea (78%) respiratory symptoms (67%) fever and night sweats (56%) and central nervous system symptoms (50%). It is believed that more than 50% of prostitutes in the Ivory Coast are from Ghana. Many of these women return to their home country for holidays or to seek medical care when they develop symptoms of HIV infection. Unless effective intervention is implemented to limit the heterosexual spread of AIDS in Ghana its incidence may soon become as high as it is in Central Africa.

In gene expression analyses using a high-density oligonucleotide array in a rat ischemia model, two comparison methods, "pair-wise comparison" and "sample average comparison", were evaluated based on statistical methods. The reliability of the elements screened with a 1.2 to 10-fold threshold was also evaluated. In pair-wise comparisons, most of the elements were significantly independent of the threshold value, with the percentage of significant elements remaining above 95%, when screened at 2.5-fold or higher threshold value. Pair-wise comparison structurally provided strict screening, which resulted in genes that were not selected despite significant alterations in expression. Screening by "sample average comparison" resulted in elements with low probability of significance, which suggested the necessity for increasing the reliability by additional statistical analyses after screening. When genes with altered expression were screened using an oligonucleotide array, marked differences in the numbers and reliability were proved to exist among elements screened by each sample comparison method.

Liver metastasis of colorectal malignancies is an important prognostic factor. Several studies have demonstrated that carbon dioxide (CO2) pneumoperitoneum enhances liver metastasis in animal models. Little is known about intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha (TNF-(alpha) mRNA expression in the liver after CO2 pneumoperitoneum.Forty-five male BALB/c mice were randomly divided into three groups after intra-splenic tumor cell (colon 26) inoculation and the following procedures were performed: CO2 pneumoperitoneum (n = 15), open laparotomy (n = 15), and anesthesia alone (n = 15). On day 7 after each procedure, the livers were excised and the number and diameter of the tumor nodules and the cancer index score were determined. Another 90 male BALB/c mice were randomly divided into three groups as described above, and they underwent each procedure (n = 30 each). After each procedure, the livers were excised on days 0, 1, 3, and ICAM-1 and TNF-alpha mRNA expression were examined by real-time RT-PCR using SYBR Green I.The number of tumor nodules and the cancer index score were larger in the CO2 pneumoperitoneum group than in the control group (p0.05). The mean diameter of the tumor nodules was not different among the three groups. The expression of ICAM-1 in the CO2 pneumoperitoneum group was higher than that in the other groups on day 1 (p0.05), and the TNF-alpha mRNA was higher than that in the control group on day 1 (p0.05).CO2 pneumoperitoneum enhances liver metastasis compared with anesthesia alone, and ICAM-1 expression in the liver after the pneumoperitoneum plays an important role in establishing liver metastasis in a murine model.

Liver metastasis is an important prognostic factor in advanced colorectal cancer. Several studies have demonstrated that carbon dioxide (CO2) pneumoperitonem enhances liver metastasis in an animal model. In the present study, we used scanning electron microscopy (SEM) to investigate morphological changes in hepatic vascular endothelium after CO2 pneumoperitoneum in a murine model. Thirty-three male BALB/c mice were randomized to undergo pneumoperitoneum (CO2, air, or helium ), open laparotomy, and anesthesia alone. After each procedure, the animals’ livers were excised at days 0, 1, and 3 and examined by SEM. In the CO2 pneumoperitoneum group, we observed rough surface and derangement of the hepatic vascular endothelial cells and intercellular clefts on day 1. In the other groups, no major morphologic changes were observed at any time. Hepatic vascular endothelium changes after CO2 pneumoperitoneum. Such characteristic changes may play an important role in establishing liver metastasis after CO2 pneumoperitoneum.

We investigated the expression of 50 cytochrome P450 (CYP) genes in the post-ischemic reperfused rat liver using a DNA microarray. Thirteen CYPs showed absent expression in all experiments and 2 CYPs were induced by pentobarbital, which was used as an anesthetic. Eight of the remaining 35 CYPs showed significantly decreased expression following ischemia-reperfusion. Monitoring CYP expression may be a powerful approach for elucidation of pathways regulating drug metabolism that may be involved in postischemic reperfusion liver injury. Keywords:: cytochrome P450, postischemic liver reperfusion, gene expression

Background: Accurate staging of lymph node metastasis by sentinel node biopsy is easily achieved in conventional open gastric surgery. Staging is not easily achieved in laparoscopic surgery, however, because of the technical difficulty in identifying sentinel nodes. We developed a laparoscopic method that involves lead shielding for detection of sentinel nodes in gastric cancer and examined the efficacy of this method. Methods: Laparoscopic sentinel node biopsy was performed in 18 patients with early gastric cancer. A combined dye- and radio-guided method was used in the first 10 patients; our radio-guided lead shield method was used in the subsequent eight cases. Laparoscopy-assisted distal gastrectomy was performed in all patients, and dissected nodes were examined by routine hematoxylin and eosin staining. The detection rate was compared between the two groups. Results: The detection rate for sentinel nodes was higher with the radio-guided lead shield method (88%) than with the combined dye- and radio-guided method (40%). Regional lymph node metastasis was recognized in one of 18 patients, and the sentinel node was positive in this case. Conclusions: Use of a lead shield is beneficial for accurate laparoscopic detection of sentinel lymph nodes.

From conventional relative gene expression analyses (Northern blotting, in situ hybridization, and RT-PCR), it has been reported that the expression of control genes, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin, used as references may be affected by ischemia. Therefore, we extended searching and evaluation at the mRNA level of transcripts whose expression levels were not changed by cerebral ischemia, using a high-density oligonucleotide array and statistical analysis in a rat global cerebral ischemia and reperfusion model. We added a hyperthermic factor and localization factor to ischemia and identified transcripts with a stable expression level under conditions even more disadvantageous than ischemia only. Screening of more than 8,000 transcripts with the Rat Genome U34A array yielded 28 transcripts, which we listed and classified according to their expression level. Widely used control genes, GAPDH and β-actin, were not included, although cyclophilin A was included. In addition, we conducted a functional classification based on gene ontology. Under the functional classification of the 28 transcripts, many genes tended to be associated with metabolism. In conclusion, use of several transcripts is recommended, such as those we identified, as references in the analysis of gene expression in pathological models of ischemia. © 2004 Wiley-Liss, Inc.

We report herein a case of primary early ileal adenocarcinoma diagnosed definitively by colonoscopy before surgery and treated by laparoscopy-assisted ileocecal resection. A 66-year-old man visited our hospital because of fecal occult blood. Colonoscopy revealed a relatively clearly demarcated, flat elevation in the terminal ileum; it was slightly red, surrounding a hemorrhagic spot, and had a small depression at its center. The lesion was diagnosed on biopsy as a well-differentiated adenocarcinoma. Laparoscopy-assisted ileocecal resection was performed. Macroscopically, the resected specimen appeared as a partially elevated, partially depressed, reddish lesion on Kerckring's fold. It was 1.5 × 1.0 cm and located 3.5 cm from the ileocecal valve. Microscopically, the tumor was diagnosed as a type IIa + IIc well-differentiated adenocarcinoma; it was confined to the mucosa, and lymph node metastasis was not found. The patient's postoperative course was uneventful, and he is now alive, well and showing no evidence of recurrence 1 year after surgery. Early detection of ileal cancer depends on careful colonoscopic observation of the terminal ileum, where small bowel cancer usually locates, even during routine examination. To our knowledge, this is the first report of laparoscopic surgery for preoperatively diagnosed early ileal carcinoma.

The patient was an 81-year-old man who was admitted to the Arita Gastrointestinal Hospital for anemia. Upper gastrointestinal series and endoscopic examinations revealed transpyloric prolapse of a pedunculated polypoid gastric carcinoma with an abnormal mucosal fold from the posterior wall of the lesser curvature of the gastric antrum. Biopsy and histological examination led to a diagnosis of papillary adenocarcinoma. The incarcerated tumor could not be restored endoscopically into the stomach, the lesion head was ulcerated, and the serum carbohydrate antigen (CA) 19-9 was elevated to 379.7 U/mL. Surgery was indicated. Intraoperatively, the tumor was found in the duodenal bulb; it was easily manipulated into the stomach, and distal gastrectomy was performed. The tumor, 5.0 × 4.2 × 2.5 cm, was diagnosed histologically as papillary adenocarcinoma of the stomach infiltrating into the submucosal layer. The carcinoma cells stained positively for CA19-9 antibody. The postoperative course was uneventful, the anemia improved, and the serum CA19-9 level decreased to within normal range. The patient remains alive 15 months after the surgery. This may be the first report of a patient with Type 0 I early gastric cancer characterized by both CA19-9 production and transpyloric prolapse.

Objective:TP53 mutations are the most frequent genetic alterations in colon cancer. We studied whether the recently developed oligonucleotide microarray technique, GeneChip p53 assay, can be applied to sensitive detection of TP53 gene mutations in surgical specimens from colon cancer patients. Methods:TP53 gene mutations in exons 2–11 in 20 colon cancers and the corresponding histopathologically normal mucosa at the surgical margins were assessed by GeneChip p53 assay, and the results were further evaluated by direct sequencing of the involved exon or by mutant-allele-specific amplification (MASA). The expression of TP53 protein was also evaluated immunohistochemically and the result was compared with the gene alteration. Results: The GeneChip p53 assay detected TP53 mutations in 65% of primary cancers; 61% of the mutations were within the evolutionarily conserved regions, and 46% of the mutations were within the zinc-binding domains (regions of loop 2 and loop 3). Direct sequencing confirmed these mutations. Immunohistochemical examination detected TP53 protein overexpression in 47% of primary cancers, but this protein did not accumulate with all types of TP53 mutations. In addition, the GeneChip assay detected a mutation identical to that in the primary tumor in 2 samples from the surgical margins, and MASA confirmed both mutations, implying the presence of occult cancer cells. Conclusion: The GeneChip p53 assay is sufficiently sensitive to detect TP53 mutations in surgical specimens from colon cancers and may be applicable to screening examination in clinical laboratories as a routine procedure.

We report a rare case of four separate gastric ruptures resulting from blunt abdominal trauma that were successfully repaired by primary closure. A 22-year-old man injured in a motorcycle accident was admitted to our hospital where physical and radiological examinations confirmed the need for abdominal exploration. Laparotomy revealed four full-thickness lacerations in the anterior wall of the stomach. The lacerations were repaired primarily by a two-layer closure. The patient recovered gradually, and was discharged on hospital day 41, since when he has remained well. Although there has been no previous report of as many gastric lacerations following blunt abdominal trauma, surgeons should be aware of the possibility of multiple ruptures, which can be managed by simple closures.

Background:13CO2 is decreased in patients with end-stage liver disease by the [1-13C]-phenylalanine breath test. Decreased 13CO2 is supposed to be caused by the decreased ability of the liver to oxidize phenylalanine. However, no direct evidence has been reported. Methods: The [1-13C]-phenylalanine breath test was performed in galactosamine hepatitis rats (n = 14) and control rats (n = 8). Plasma phenylalanine concentration before intravenous administration of [1-13C]-phenylalanine, the elimination rate of phenylalanine and the phenylalanine hydroxylase (PAH; EC 1.14.16.1) activity of the whole liver were examined. Results: Increase of 13CO2 in the breath [Δ13CO2 (‰)] of galactosamine hepatitis rats 2 min after administration of [1-13C]-phenylalanine was only 1/5 of that in control rats. The concentration of plasma phenylalanine and the elimination rate of plasma phenylalanine in hepatitis rats did not show significant differences compared to control rats. On the other hand, a clear difference in the activity of PAH was observed between hepatitis rats and control rats. Δ13CO2 (‰) 2 min after administration of [1-13C]-phenylalanine was highly correlated to the PAH activity of the whole liver (r = 0.917). Conclusion: It was strongly indicated that decreased Δ13CO2 in hepatitis rats was the result of decreased activity of PAH.

Using a dialysis electrode, previous studies showed a clear biphasic release of glutamate during anoxia and ischemia. In this study, we examined two hypotheses: (1) glutamate is of vesicular origin and its release is thus Ca2+- and ATP-dependent in the first phase, while in the second phase glutamate is derived primarily from the metabolic pool, and (2) reversed glutamate uptake, due to electrogenic stoichiometry, produces the second phase during anoxic insult in the rat brain. A dialysis electrode continuously perfused with glutamate oxidase and ferrocene-conjugated bovine serum albumin (BSA) optimized the time resolution of monitoring, allowing quantitative oxygen-independent, real-time measurement of the extracellular glutamate concentration ([Glu]e) during anoxia. [Glu]e dynamics were analyzed during anoxia by combining the dialysis electrode with focal microinjection of substances inducing glutamate release. Following anoxia in the rat brain, a sharp and rapid [Glu]e elevation took place (first phase). The [Glu]e elevation then shifted, continuing a gently sloping rise throughout the anoxic period (second phase). This first phase disappeared with intracranial administration of either Co2+ or omega-conotoxin. The second phase rise increased with focal microinjection of KCl (300 mM, 1 microL) and decreased with NaCl (300 mM, 1 microL), ultimately reaching a plateau in both cases. Preloading with a novel glutamate transporter inhibitor (tPDC) decreased both the first and second phases of [Glu]e elevation. This dialysis electrode system provides data supporting in vivo evidence that the peak of the first phase of [Glu]e elevation is derived from the "neurotransmitter pool," while the second phase is derived from the neuronal and glial "metabolic pool," which is, at least, partly related to a "reversed uptake" mechanism in the anoxic rat brain.

Numerous reports have suggested that anoxic depolarization is a critical event in the pathogenesis of cerebral ischemia. Extracellular glutamate concentration ([Glu]e) is closely related to the pathogenesis of ischemia. Therefore, these pathogenic mechanisms merit study, especially the relationship between [Glu]e elevation and the ionic basis of early changes in membrane potential after ischemic insult in vivo. It is often presumed from electrophysiological studies that a causal relationship exists between impaired glutamate uptake and/or progressive glutamate increase and anoxic depolarization, but few in vivo reports have found any sign of a progressive increase of [Glu]e elevation preceding anoxic depolarization. Recently, we reported the application of an oxygen-independent real-time technique for monitoring glutamate levels in the extracellular space during in vivo ischemia, and demonstrated that the massive glutamate release during ischemia is biphasic. In the present study, using this real-time monitoring system, we carried out a more detailed analysis of the initial events in the first phase of glutamate release during ischemia-induced anoxic depolarization. The shape of the rising slope that forms the peak of the first phase suggested two components. The second component was approximately 10 times steeper than the first, with two different components of the rise on the way to the peak of the biphasic [Glu]e elevation. This is the first report to demonstrate these components of the initial glutamate peak, and suggests a progressive second component of the [Glu]e increase preceding Ca2+-dependent release from synaptic vesicles with anoxic depolarization, in vivo.

The biological behavior of neuroblastomas detected through mass screening (MS,/=1 year of age) and that of mass screening-negative later-presenting (MSN,1 year of age) neuroblastomas have been reported to differ in many studies. To investigate the biological differences between these two groups, we analyzed the differences in mRNA profiles.We analyzed the mRNA profiles of MS and MSN neuroblastomas using differential display, and cloned and sequenced the bands differentially expressed between these two groups. Using the RNA analysis by polymerase chain reaction (RNA-PCR) method, the relative amount of mRNA in tumor tissue in each sample was measured. Associations between relative amount of mRNA and clinical and genetic variables related to patient prognosis and the effect of the level of mRNA expression on survival probability were investigated using statistical methods.Using differential display and RNA-PCR, we found that the mRNA for the human homologue of the yeast cdc10 gene (hCDC10) identified in Saccharomyces cerevisiae was expressed at a higher level in the MS group of patients than in the MSN group of patients (0.554 +/- 0.197 for MS neuroblastoma, n = 24 and 0.244 +/- 0.179 for MSN neuroblastoma, n = 10, P0.01), and this difference was suggested to be independent of the histologic subtype of tumor. A high level of hCDC10 mRNA expression in neuroblastomas (relative amount of hCDC10 mRNA0.35) was also suggested to be associated with younger age at diagnosis (/=1 year of age, P0.01), favorable clinical stage (I, II, and IVs, P0. 01), and favorable histology in the Shimada classification (P0. 01), whereas a low level of hCDC10 mRNA expression (relative amount of hCDC10 mRNA/=0.35) was suggested to be associated with the progression of clinical stage (P0.01) and N-myc gene amplification (1 copy, P0.05). Patients with neuroblastomas with a high level of hCDC10 mRNA expression were suggested to have a better prognosis than those with a low level of hCDC10 mRNA expression (P0.01).A high level of hCDC10 mRNA expression in neuroblastomas may be associated with favorable clinical and biological characteristics, and the expression of hCDC10 mRNA in neuroblastomas may affect the clinical and biological characteristics of this type of tumor.

Changes in brain temperature are known to modulate the marked neuronal damage caused by an approximately 10-min intra-ischemic period. Numerous studies have suggested that the extracellular glutamate concentration ([Glu](e)) in the intra-ischemic period and the initial postischemia period is strongly implicated in such damage. In this study, the effects of intra-ischemic brain temperature (32, 37, 39 degrees C) on [Glu](e) were investigated utilizing a dialysis electrode combined with ferrocene bovine serum albumin (BSA), which allows oxygen-independent real-time measurement of [Glu](e). This system allowed separate quantitative evaluation of intra-ischemic biphasic glutamate release from the neurotransmitter and metabolic pools, and of postischemic glutamate re-uptake in ischemia-reperfusion models. The biphasic [Glu](e) elevation in the intra-ischemic period did not differ markedly among intra-ischemic brain temperatures ranging from 32 to 39 degrees C. Intra-ischemic normothermia (37 degrees C) and mild hyperthermia (39 degrees C) markedly inhibited [Glu](e) re-uptake during the postischemic period, although the intra-ischemic [Glu](e) elevation did not differ from that during intra-ischemic hypothermia (32 degrees C). It was assumed that normothermia or mild hyperthermia in the intra-ischemic period influences intracellular functional abnormalities other than the intra-ischemic [Glu](e) elevation, thereby inhibiting glutamate re-uptake after reperfusion rather than directly modulating intra-ischemic [Glu](e) dynamics.

We simultaneously measured extracellular glutamate ([Glu]e) elevation and local CBF using a real-time monitoring method and laser-Doppler flowmetry, respectively, in the rat striatum in a modified graded global ischemia model. Ischemic brain temperatures were kept at 32 degrees C, 37 degrees C and 39 degrees C. Three distinct types of intraischemic [Glu]e elevation, reflecting mild, moderate and massive glutamate release, were observed. Brain temperature plays an important role in determining CBF thresholds for each of the three types of [Glu]e elevation. CBF thresholds for [Glu]e elevations shifted to a lower level range as brain temperature was reduced. In mild or moderate ischemia, there is no exposure to sustained [Glu]e elevation, which is seen only in relatively severe ischemia characterized by biphasic [Glu]e elevation.

Objective In recent years, small high-performance batteries have become very popular. With this increasing miniaturization of batteries, clinicians have noted an increasing frequency rate of esophageal injury due to battery ingestion by infants. The sltuation is associated with severe injury to the esophagus due to the electrical current produced, particularly in the case of high-performance batteries producing high currents. The pathophysiologic features and complications of esophageal battery burns have not been thoroughly investigated. Our study intended to investigate the pathophysiologic features and complications of esophageal battery burn. Design Open, randomized, controlled study. Setting Experimental animal laboratory in a university hospital. Subjects Male adult mixed-breed rabbits, 22 wks old and weighing 3 to 3.5 kg. Interventions The experimental rabbit model of esophageal injury due to battery ingestion described herein was designed to study not only the direct influence of contact with the battery but also damage to neighboring tissues and the biochemical and pathologic mechanisms of injury. We investigated the relationship between the direction of the inserted battery and the mechanism underlying these complications. Esophageal burn injury was created by placing a 3-V battery into the esophagus for 9 hrs. Measurements and Main Results The cathode side of the esophagus became increasingly alkaline, while the anode side was acidic. Low-voltage battery burns are likely to be due to secondary chemical reactions caused by the electric current because of acid generated at the anode and alkall at the cathode using a micro pH meter. Injury was significantly more severe on the alkaline side when a battery was placed with its cathode directed toward the trachea. Alkaline complications affecting neighboring tissues were more severe than acid complications. These results indicate that as well as the esophageal mucosa itself being injured, deleterious effects are exerted on surrounding tissues, the severity of which vary depending on the orientation and duration of the battery being lodged in the esophagus. Conclusions The direction of the battery cathode, which produces alkali, is important in determining the severity of complications. Based on our investigation of the underlying mechanisms of these complications, we advocate the establishment of treatment guidelines for battery swallowing accidents. (Crit Care Med 1997; 25:2039-2044)

Analyzing the distribution pattern of transferrin (Tf) and ferritin, we investigated the changes in iron metabolism related proteins in the process of neuronal death induced by 5 min ischemia. In the control animals, Tf immunoreactivity was localized in the oligodendrocytes. Ferritin was distributed in both neurons and gliacytes, particularly microglia. In parallel with the delayed neuronal death, Tf-positive atrophied neurons and numerous ferritin-positive gliacytes appeared in the CA1 subfield of the hippocampus 4 days after ischemia, when glia fibrillary acidic protein (GFAP)-positive astrocytes also appeared throughout the hippocampal structure. A considerable number of ferritin-positive phagocytes (reactive microglia) appeared in the stratum pyramidale from the seventh day. Our data show clearly that the mobilization of Tf and ferritin-positive phagocytes are linked with the degeneration of neurons induced by cerebral ischemia. These events may suggest an activation of iron handling system under the postischemic condition.

Glutamate has been proposed to play a critical role in acute ischemic pathophysiology in the brain. In this study, glutamate was monitored by the dialysis electrode technique, in which glutamate is oxidized by glutamate oxidase producing hydrogen peroxide which is then amperometrically detected on a platinum electrode set at +650 mV vs Ag/AgCl. A dialysis electrode, which consists of a microdialysis probe with a built-in platinum electrode, provides a continuous glutamate oxidase perfusion inside of the probe. Perfusion with this solution allows real-time monitoring of glutamate dynamics in the extracellular space during ischemia. This study was designed to collect detailed information on rapid changes in the extracellular glutamate concentration of the rat striatum and demonstrated two distinct phases of glutamate release during early severe brain ischemia.

The distribution patterns of cystatin C and apolipoprotein E (apo E) were studied immunocytochemically in the gerbil hippocampus before and after 5 min ischemia. In the controls, cystatin C was distributed mainly in astrocytes. In addition, a large number of dots positive for cystatin C were observed around the outlines of neuronal perikarya in the CA1 subfields. One day after ischemia, cystatin C-positive stainings outlining neuronal cell bodies disappeared. On the fourth day, intense stainings for cystatin C appeared in atrophied pyramidal neurons and these stainings in neurons disappeared by the 14th day. A remarkable increase in the number of cystatin C-positive astrocytes occurred on the fourth day and thereafter these spread over the whole of the CA1 subfield. Apo E was also distributed in astrocytes in the control specimens. From the fourth day, extra- and/or intracellular distribution of apo E-immunoreactivities was noted in the stratum pyramidale. Apo E-positive astrocytes disappeared transiently on the fourth day and then reappeared and increased remarkably by the 14th day. These findings indicate that cystatin C and apo E are involved in the degeneration process of brain neuronal cells.

Discoveries of long-term potentiation and immediate early gene in the central nervous system have enabled new developments in experiments on learning and memory. These experiments are conducted in many kinds of animals with different procedures, physiology, chemistry and pharmacology. However, there is still some confusion when these various procedures are discussed. Memory is defined as information storage of an animal's previous experiences. The memory induces changes in behavioral performance. This means that memory must be observed in whole animals, and one question that can occur is how does long-term potentiation, for example, correlate with memory. Furthermore, memory has been divided into two major classifications, declarative and non-declarative, from the comparison of amnesias observed in humans and animals. The declarative memory can be observed in human subjects, but not in animals. This article presents a neuronal circuit concerning memory formation and some results obtained from benzodiazepines, and it discusses some problems encountered executing when experiments on learning and memory. In addition, the discussion speculates over the possibility for an "anti-dementia drug".

720 Background: Fluorouracil-based chemoradiotherapy (CRT) is regarded as a standard perioperative treatment in locally advanced rectal cancer. We investigated the efficacy and safety of substituting fluorouracil with the oral prodrug S-1. Methods: A multi-institutional (17 specialized centers), interventional phase II trial, was conducted from April 2009 to August 2011. For inclusion, patients must fulfill the following requirements before neoadjuvant CRT: (i) histologically proven rectal carcinoma; (ii) tumor located in the rectum (upper, lower); (iii) cancer classified as T3-4, N0–3 and M0; Two cycles of neoadjuvant CRT with S-1 (100 mg/m2 on days 1-5, 8-12, 22-26, and 29-33) was administered, and irradiation (total 45Gy/25fr, 1.8Gy/day, on days 1-5, 8-12, 15-19, 22-26, and 29-33) was performed. Total mesorectal excision was performed during the 4th and 8th week after the end of the neoadjuvant CRT. The primary endpoint is rate of complete treatment of neoadjuvant CRT. Secondary endpoints are response rate of neoadjuvant CRT, short-term clinical outcomes, rate of curative resection, and pathological response (grade2/3). Results: This trial included 37 patients (clinical StageIIA: 8, IIIB: 19, IIIC: 10; tumor located in the upper rectum; 4, the lower rectum; 33). A complete treatment of neoadjuvant CRT was found in 86.5% of patients (95%CI;75.5-97.5%), and an adverse event (grade 3/4) occurred in 4 patients(11.1%). Response rate (PR/CR;RECIST 1.0) was 56.8% (95%CI; 40.8-72.7%), and pathologic response rate (grade2/3) was 48.6% (95%CI; 32.5-64.8%). The median operating time was 448.5 min (IQR 340.5-505.5), and median blood loss was 422.5 mL (IQR 182.5-1125). Grade 3-4 postoperative complications occurred in 6 (16.7%) patients. The most common grade 3 or 4 postoperative complication was anastomotic leakage (2 [5.6%]). The 3-year overall survival rate was 88.5%. The 3-year disease free survival rate was 70.9%. Median length of follow-up was 42 months. Conclusions: A neoadjuvant-synchronus S-1+Radiotherapy for locally advanced rectal cancer is feasible in terms of pathological response, adverse events, accompany with favorable long-term outcome. Clinical trial information: 03396.

INTRODUCTIONIdiopathic thrombocytopenic purpura (ITP) is an autoimmune disease characterized by a low platelet count and normal bone marrow. Patients with ITP undergoing surgery are thought to have increased risk for postoperative complications because of their thrombocytopenia.PRESENTATION OF CASEwe report the case of a 66-year-old woman with ITP who required an emergency operation for acute appendicitis associated with disseminated intravascular coagulation. Preoperative therapy consisted of platelet transfusions only, and intraoperative hemostasis was achieved. Postoperatively, high-dose intravenous immunoglobulin (IVIg) therapy led to an increased, stable, and adequate platelet count and good hemostasis.DISCUSSIONThe outcome of this case suggests that IVIg therapy is not always required for preoperative management of patients with.CONCLUSIONIVIg therapy may be useful for postoperative management after emergency surgery.

Background Buruli ulcer (BU) caused by Mycobacterium ulcerans (M. ulcerans) has emerged as an important public health problem in several rural communities in sub-Saharan Africa. Early diagnosis and prompt treatment are important in preventing disfiguring complications associated with late stages of the disease progression. Presently there is no simple and rapid test that is appropriate for early diagnosis and use in the low-resource settings where M. ulcerans is most prevalent. Methodology We compared conventional and pocket warmer loop mediated isothermal amplification (LAMP) methods (using a heat block and a pocket warmer respectively as heat source for amplification reaction) for the detection of M. ulcerans in clinical specimens. The effect of purified and crude DNA preparations on the detection rate of the LAMP assays were also investigated and compared with that of IS2404 PCR, a reference assay for the detection of M. ulcerans. Thirty clinical specimens from suspected BU cases were examined by LAMP and IS2404 PCR. Principal Findings The lower detection limit of both LAMP methods at 60°C was 300 copies of IS2404 and 30 copies of IS2404 for the conventional LAMP at 65°C. When purified DNA extracts were used, both the conventional LAMP and IS2404 PCR concordantly detected 21 positive cases, while the pocket warmer LAMP detected 19 cases. Nine of 30 samples were positive by both the LAMP assays as well as IS2404 PCR when crude extracts of clinical specimens were used. Conclusion/Significance The LAMP method can be used as a simple and rapid test for the detection of M. ulcerans in clinical specimens. However, obtaining purified DNA, as well as generating isothermal conditions, remains a major challenge for the use of the LAMP method under field conditions. With further improvement in DNA extraction and amplification conditions, the pwLAMP could be used as a point of care diagnostic test for BU, Author Summary In order to develop a simple and rapid test that can be used to diagnose Buruli ulcer under field conditions, we modified the conventional LAMP assay by using a disposable pocket warmer as a heating device for generating a constant temperature for the test reaction and employed the use of crude sample preparations consisting of boiled and unboiled extracts of the clinical specimen instead of using purified DNA as the diagnostic specimen. Thirty clinical specimens from suspected Buruli ulcer patients were investigated by the modified LAMP (or pocket warmer LAMP) and the conventional LAMP, as well as IS2404 PCR, a reference method for the detection of Mycobacterium ulcerans. There was no significant difference in the detection rate (63–70%) in all of the methods when purified samples were used for the tests. On the other hand the use of crude specimen preparation resulted in a drop in detection rate (30–40%). This study demonstrates that the LAMP test can be used for rapid detection of M. ulcerans when purified DNA preparations are used. With further improvements in the sample reaction, as well as in specimen purification, the pocket warmer LAMP may provide a simple and rapid diagnostic test for Buruli ulcer.

In order to determine the role of excitatory amino acids (EAAs) in free fatty acid (FFA) liberation during cerebral ischemia, we examined the effect of in situ administration of kynurenic acid, a broad-spectrum antagonist of EAA receptors, by microdialysis on the increase in FFA levels during ischemia in the rat hippocampus. A transient rapid increase in FFA levels, superimposed on a continued slow increase, was observed beginning at 1-2 min after ischemia induction. The early rapid increase in FFAs was profoundly inhibited by kynurenic acid, suggesting that EAAs are critically involved in the early phase of FFA liberation. Development of massive ionic shifts during cerebral ischemia can be delayed for several minutes by kynurenic acid administered by the same procedure, suggesting a vital role for EAAs in the early appearance of anoxic depolarization. The observed inhibition of early FFA liberation may thus be attributable to the delay in development of massive ionic shifts and resultant neurotransmitter release which may activate phospholipase A2 and C.

Transfusion-transmissible infections among 808 blood donors in Ghana were investigated in 1999. Antibody seroprevalences of 3.8, 0.7, 8.4, and 13.5%, respectively, for human immunodeficiency virus, human T-cell lymphotrophic virus type 1, hepatitis C virus (HCV), and Treponema pallidum were obtained. The seroprevalence of HCV infection was confirmed to be 0.9% after supplementary testing, and the transfusion risk potential of these pathogens was demonstrated.

BACKGROUND The Diagnosis Procedure Combination (DPC) is a national administrative case-mix classification system for acute inpatient care which was launched in Japan in 2003. This system was designed to increase the standardization and transparency of medical information. The system has collected medical information in a unified format nationwide from acute hospitals. Significant parts of the data has been published by Ministry of Health, Labour and Welfare, including average length of hospital stay and number of patients by disease and hospital. This information is valuable for medical consultation in occupational health settings. OBJECTIVES This article provides an overview of the DPC system and how to utilize the data from the perspective of occupational health practitioners.

As recent technological innovations make it possible to clarify the concordant relationship between genomic alterations and aberrant gene expression during the progression of colorectal cancer (CRC), we aimed at identifying new overexpressing genes with genomic amplification on the responsible loci in CRC. The candidate gene was found using cDNA microarray and array-based comparative genomic hybridization (CGH) analysis after laser microdissection (LMD) in 132 Japanese CRC. We focused on SUGT1, which is associated with the assembling of kinetochore proteins at the metaphase of the cell cycle, with significant association between genetic alterations and expression. SUGT1 mRNA expression was evaluated in 98 CRC cases to determine the clinicopathological significance of SUGT1 expression. The mean level of SUGT1 mRNA expression in tumor tissue specimens was significantly higher than in non-tumor tissue. The high SUGT1 expression group was characterized by a significantly elevated frequency of recurrence and a significantly poorer prognosis than the low expression group. There was a significant association between poor prognosis of CRC cases and the overexpression of SUGT1 with genomic amplification of the loci concordantly. The amplification of SUGT1 might give rise to promote the transcription of the gene directly subsequent to the progression of CRC cases with worsening prognosis.

Background Little is known about the relation between HLA-I expression and the prognosis of patients with gastric cancer. The aim of this retrospective study was to clarify the clinical significance of HLA-I heavy chain expression in gastric cancer. Methods The study subjects were 202 patients with gastric cancer who had undergone curative surgery. Tumors were examined for expression of HLA-I heavy chain antigens by immunohistochemistry. We analyzed the association of HLA-I heavy chain expression with clinicopathological parameters and patient prognosis. Results HLA-B/C expression showed association with deeper tumor invasion, higher incidence of lymph node metastasis, more advanced tumor stage, and higher incidence of recurrence. Patients with positive HLA-B/C expression had shorter 5-year overall and 5-year disease-free survival compared with patients whose tumors showed mixed and negative expression (P

Mild to moderate differences in brain temperature are known to greatly affect the outcome of cerebral ischemia. The impact of brain temperature on ischemic disorders has been mainly evaluated through pathological analysis. However, no comprehensive analyses have been conducted at the gene expression level. Using a high-density oligonucleotide microarray, we screened 24000 genes in the hippocampus under hypothermic (32 degrees C), normothermic (37 degrees C), and hyperthermic (39 degrees C) conditions in a rat ischemia-reperfusion model. When the ischemic group at each intra-ischemic brain temperature was compared to a sham-operated control group, genes whose expression levels changed more than three-fold with statistical significance could be detected. In our screening condition, thirty-three genes (some of them novel) were obtained after screening, and extensive functional surveys and literature reviews were subsequently performed. In the hypothermic condition, many neuroprotective factor genes were obtained, whereas cell death- and cell damage-associated genes were detected as the brain temperature increased. At all intra-ischemic brain temperatures, multiple molecular chaperone genes were obtained. The finding that intra-ischemic brain temperature affects the expression level of many genes related to neuroprotection or neurotoxicity coincides with the different pathological outcomes at different brain temperatures, demonstrating the utility of the genetic approach.

e14612 Background: Fluorouracil-based chemoradiotherapy (CRT) is regarded as a standard perioperative treatment in locally advanced rectal cancer. We investigated the efficacy and safety of substit...

Goals: We sometimes encounter residual or recurrent cancers after endoscopic mucosal resection (EMR) for early gastric cancer. The aim of the present study was to clarify the clinicopathologic characteristics of and optimal treatment for the residual cancers after EMR. Study: Seventy-four patients with early gastric cancer were treated with EMR between 1994 and 2004. These patients were divided into 2 groups as follows: the curative group (n = 59) and the noncurative group (n = 15). The clinicopathologic data were compared between the 2 groups and the outcomes of additional therapy were reviewed. Results: In the noncurative group, the tumors were located significantly frequently on the upper or middle third of the stomach compared with the curative group (P < 0.05). The number of fragments in EMR was significantly larger in the noncurative group than in the curative group (P < 0.05). Fifteen patients required additional treatment because of the residual cancer. Nine (75%) of 12 patients requiring surgery underwent laparoscopic surgery. Three patients were treated by endoscopic therapy. Conclusions: EMR with a single fragment and with a sufficient margin is useful for the complete resection of early gastric cancer. When residual cancer occurs, laparoscopic gastrectomy may be a good alternative.

A 33-year-old woman, gravida 2, para 1, was diagnosed to have a benign mucinous cystic neoplasm of the pancreas 5 months before delivery. The tumor measured 12 cm in diameter at the time of diagnosis. The antenatal course was uneventful, and a vaginal delivery produced a normal infant. By 2 months after delivery, the tumor reached 18 cm. At surgery, a huge cyst was found to originate from the pancreas, and a distal pancreatectomy with splenectomy was performed. The cystic mass was multilocular 18 x 17 x 12 cm, 2450 g, and red to yellowish-gray. The histologic diagnosis was benign mucinous cystadenoma. The postoperative course was uneventful, and the patient remains free of recurrence at 7 months after surgery. To our knowledge, this is only the fifth reported case of pancreatic mucinous cystadenoma in association with pregnancy. This is the first reported case of a successful resection of such a tumor after delivery.

Hepatitis B virus (HBV) genotypes from A to H have distinct geographical distributions and have been shown to affect the clinical features as well as the course of the HBV infection. HBV genotype E has been found only in Africa. However, the complete genomes of this genotype, which were isolated mainly from West Africa, were available only for a few samples. In this study, five full-length genomes and seven other small S genes of HBV strains from Ghanaian blood donors were sequenced and investigated. Following phylogenetic analysis, all of the Ghanaian HBV strains were clustered closely in genotype E. All of the 12 small S genes showed the same characteristic of subtype ayw4. The complete genomes of the five Ghanaian strains showed marked similarity with each other and with the reported genotype E strains (96.7%-99.1%). Genotype E strains showed low intra-genotypic diversity (1.8%) and carried the conserved signature pattern in pre-S1 as well as in the full genome sequence. Of note, the finding of the G145R escape mutant in an unvaccinated Ghanaian blood donor might raise concern as to the ongoing nation-wide hepatitis B vaccination program in Ghana.

Goals: Early mucinous gastric carcinoma (MGC) is very rare, and the clinicopathologic features are not well understood. The aim of this study is to clarify the clinicopathologic characteristics of early MGC. Study: A total of 806 patients with gastric cancer underwent gastrectomy between 1983 and 2002 at our department. Of these patients, 7 were found to have early MGC. MGC was defined as a tumor in which more than 50% of the tumor area contained extracellular mucin pools. Microscopic examination was performed, and particular attention was paid to the mucin component of the tumor and histologic subtype (ie, well-differentiated or poorly-differentiated). Results: The incidence of early MGC among all gastric carcinomas in our series was 0.9% (7/806). Tumors ranged in size from 1.0cm to 7.2 cm, with a mean of 3.7 cm, and 4 appeared macroscopically elevated and 3 appeared depressed. Histologically, 3 tumors were considered well-differentiated type and 4 were considered poorly differentiated type. Microscopic features were thickening of the submucosal layer due to accumulation of abundant mucin and elevation of the surrounding normal mucosa. Tumor invaded the submucosa in 6 cases, and lymph node metastasis occurred in 1 case. However, no patient died of recurrence during follow-up periods ranging from 4 to 85 months. Conclusions: Early MGC is characterized as an elevated lesion resembling submucosal tumor due to abundant mucin pools in the submucosa. Cases of early MGC have a good outcome similar to that of early non-MGC.

Lymph node metastasis is one of the most important prognostic factors for cancer patients. There are few animal models of lymph node metastasis. The purpose of this study was to develop a simple animal model without surgical trauma. B16F10 melanoma (1 x 10(6) cells in 0.1 ml phosphate-buffered saline) were slowly injected into the retroperitoneal space followed by direct puncture at the position between the anus and tail of 50 female C57BL6 mice. At 1-week intervals for 4 weeks after the procedure, we examined the retroperitoneal tumor and intra-abdominal lymph nodes. There was no morbidity and no mortality. At 2 weeks after inoculation, the retroperitoneal tumor was visible macroscopically at the position between the rectum and the sacrum, and histological examination showed the occurrence of intra-abdominal lymph node metastasis in all mice. The number of positive nodules was increased and was affected by the amount of cancer cells inoculated and the interval from inoculation to lymph node metastasis. A convenient murine model for the study of lymph node metastasis has been developed. Our animal model does not involve surgical trauma and may be useful in the analysis of the association between surgical stress and lymph node metastasis and in the elucidation of the mechanism and prevention of lymph node metastasis.

Conventional open gastrectomy has been reported to result in increased morbidity in obese patients. To date, there has been no study evaluating laparoscopic gastrectomy in such patients; therefore, we assessed the short-term results of this procedure in a group of obese patients. The study included 99 consecutive patients who underwent laparoscopy-assisted distal gastrectomy (LDG) for the cure of early gastric cancer. The patients were divided into two groups: obese (body mass index [BMI] ≥25.0, n = 16) and nonobese (BMI

Our newly developed method using a dialysis electrode has made it possible to perform real time monitoring of extracellular glutamate concentration ([Glu]e) utilizing the oxygen-independent reaction with glutamate oxidase and ferrocene. In this study, we therefore, investigated [Glu]e changes during brain ischemia using both the conventional microdialysis method and the dialysis electrode method. A comparison between our newly developed dialysis electrode and conventional microdialysis methods provided the following results. When the conventional microdialysis method was employed: (1) the elevation of [Glu]e during complete global ischemia was delayed; and (2) the elevation of concentration and reuptake of glutamate were delayed during 10-min transient ischemia, and the elevation of [Glu]e reached a maximum later using conventional microdialysis than using our dialysis electrode. (3) The biphasic [Glu]e elevation of glutamate concentration detected using the dialysis electrode method was not observed using the conventional microdialysis method. It was additionally investigated why the conventional microdialysis method provides inferior time resolution. In this study, we also demonstrated with the chromatographic SMART procedure coupled to UV detection that biogenic substances, i.e. low molecular weight proteins and peptides, are released during ischemic injury, and they may cause a delay in the time resolution in the microdialysis method.

We performed dual (two-color) fluorescence in situ hybridization (FISH) using direct fluorescent labeling probes for p53 and chromosome 17 in six gastrointestinal (3 stomach and 3 colon) cancers. In three of these (1 stomach and 2 colon) the interphase cell nuclei showed an imbalance of signals for the p53 and chromosome 17; that is, the p53 signal count was lower than the chromosome 17 signal count, indicating deletion of the p53 gene. Moreover, metaphase FISH analysis demonstrated that those nuclei actually had a chromosome 17 with deletion of the p53 gene. Interestingly, these three cases had an abnormal chromosome 17 copy number, that is, chromosome 17 aneusomy. Furthermore, to investigate the possibility of p53 mutation in tumors with an imbalance of signals for chromosome 17 and p53 per nucleus, we performed a GeneChip p53 assay which has recently been developed. GeneChip p53 assay demonstrated that a primary tumor sample from one colon cancer case had a heterozygous point mutation of CGT (Arg) to CAT (His) at codon 273 in exon 8. In addition, a sample of metastatic tumor in the liver from the same case revealed two heterozygous point mutations. One of them was the same mutation as that is the primary tumor; the other was GTG (Val) to GGG (Gly) at codon 217 in exon 6. In conclusion, we found that the combination of dual-color FISH and GeneChip p53 assay offered reliable results and important information concerning not only deletion of the p53 gene and chromosome 17 aneusomy but also p53 mutations. Using these techniques, we demonstrated that an imbalance of signals for chromosome 17 and p53 per nucleus, chromosome 17 aneusomy, and accumulation of p53 mutations had occurred during carcinogenesis and development of gastrointestinal cancers.

Whereas a 2-3 degrees C decrease in intraischemic brain temperature can be neuroprotective, mild brain hyperthermia significantly worsens outcome. Our previous study suggested that an ischemic injury mechanism which is sensitive to temperature may not actually increase the extracellular glutamate concentration ([Glu](e)) during the intraischemic period, but rather impairs the Glu re-uptake system, which has been suggested to be involved in the reversed uptake of Glu. We speculated that enhancing Glu re-uptake, pharmacologically or hypothermically, may shorten exposure to high [Glu](e) in the postischemic period and thereby decrease its deleterious excitotoxic effect on neuronal cells. In the present study, rats treated with nicergoline (32 mg/kg, i.p.), an ergot alkaloid derivative, showed minimal inhibition of the [Glu](e) elevation which characteristically occurs during the 10-min intraischemic period, while Glu re-uptake was dramatically improved in the postischemic period, when severe transient global ischemia was caused by mild hyperthermia. Moreover, the nicergoline (32 mg/kg, i.p.) treated rats showed reduced cell death morphologically and clearly had a far lower mortality. The present study suggests that the development of therapeutic strategies aimed at inhibition or prevention of the reversed uptake of glutamate release during ischemia, i.e., activation of the glutamate uptake mechanism, is a promising approach to reduce neural damage occurring in response to brain ischemia.