Clinical relevance of leukocyte-associated endotoxins measured by semi-automatic synthetic luminescent substrate method.

A newly developed semi-automatic synthetic luminescence substrate (SALS) method for measuring endotoxin was compared with the existing turbidimetric kinetic assay (TKA) using leukocyte-rich plasma to verify its usefulness. As a result, the endotoxin levels by this method were higher than that by the existing assay in most specimens, and the time required for measurement was much shorter. In addition, the leukocyte-rich plasma endotoxin level minus the plasma endotoxin levels were named leukocyte-associated endotoxin, and these levels per one leukocyte were compared. As a result, those levels were highly correlated with the endotoxin measurement levels of leukocyte-rich plasma. The correlation coefficient of SALS method was superior to the existing TKA method, the endotoxin level by this method may be close to true endotoxin levels.

Auto-brewery syndrome caused by oral fungi and periodontal disease bacteria.

BACKGROUND: Auto-brewery syndrome (ABS) is often caused by fungi in the intestinal tract. We describe a rare case of alcohol production by Candida albicans and periodontal disease bacteria in the oral cavity. CASE PRESENTATION: A man aged in his 60s had a car accident, and alcohol was detected on his breath. At the time, he exhibited alcohol overdose seizures with no alcohol consumption. We carried out a gastrointestinal endoscopy, detected esophageal candidiasis, and diagnosed ABS. His seizures continued despite using miconazole oral gel. Significant tooth decay, periodontal disease, and high C. albicans levels were observed in his oral cavity. Alcohol production was confirmed from periodontal bacteria and C. albicans cultures and alcohol-degrading enzyme functions were poor. Dental treatment and antifungal drugs reduced seizures, and improved his fatty liver. CONCLUSION: Alcohol can be produced by microorganisms in healthy individuals. Therefore, blood alcohol levels and alcohol-degrading enzyme functions should be examined in patients with unexplained liver dysfunction.

Endotoxin contamination of single-use sterile surgical gloves.

Aim: Strict endotoxin limits are enforced for implants and catheters inserted into the body. However, no standard limit has been set for single-use sterile surgical gloves. Materials & methods: Four types of gloves sold in Japan were dipped in saline and that endotoxin levels were measured. Cytokine producing activity of gloves in blood was also measured. Results: Three of the four types of gloves showed endotoxin contamination. We also confirmed an increase in cytokine production in these gloves except one glove in which anionic surfactants was found. Conclusion: The extent to which detected endotoxins enter the body during surgery is controversial, but strict endotoxin limits need to be established.

Retrospective study on the effect of tight glucose control in postoperative sepsis patients using an artificial pancreas.

Tight glucose control (TGC) using a sliding scale based on intermittent blood glucose measurements occasionally can have a fatal outcome as a result of insulin-induced hypoglycemia. The present study was undertaken to examine whether the use of an artificial pancreas to achieve TGC would be possible in postoperative patients with sepsis. The retrospective study was carried out as an exploratory study, focusing on the possibility of precise evaluation of the significance of TGC as a beneficial intervention by serological monitoring of various mediators. TGC was accomplished using an artificial pancreas (STG-22; (Nikkiso, Tokyo, Japan). The patients were divided into two groups: the TGC group (6 patients with sepsis in whom the target blood glucose level set at <150 mg/dl was attempted using the artificial pancreas), and the glucose control (GC) group (6 patients with sepsis in whom glucose control was attempted using a sliding scale; target blood glucose level was set at 200 mg/dl or lower). The mean blood glucose level was 129.7 ± 9.7 mg/dl in the TGC group and 200.9 ± 14.7 mg/dl in the GC group (P < 0.01, ANOVA). No hypoglycemia associated with the artificial pancreas was seen in any of the patients. The serum levels of S100A12 and HMGB-1 tended to decrease, and those of sRAGE tended to increase, in the TGC group. Further data collection from a larger number of cases would be expected to allow a precise assessment of TGC as a potentially beneficial intervention in sepsis patients.

Significance of measuring S100A12 and sRAGE in the serum of sepsis patients with postoperative acute lung injury.

BACKGROUND: There is a report that S100A12 is useful as an early marker of acute lung injury (ALI). The purpose of this study was to determine whether S100A12 or sRAGE is useful as a marker during the development of ALI in postoperative sepsis patients. METHODS: The subjects were patients who underwent emergency surgery because of sepsis secondary to perforation of the lower gastrointestinal tract. We conducted a retrospective study comparing 2 groups of patients: a group of 9 patients who developed postoperative ALI, the ALI(+) group, and a group of 8 patients who did not develop postoperative ALI, the ALI(-) group. Their blood S100A12, sRAGE, IFN-gamma, WBC count, and CRP values were measured immediately after surgery and on postoperative day 1 (D1). RESULTS: The changes in S100A12 showed significantly higher values immediately postoperatively in the ALI(+) group (p < 0.05). The sRAGE values immediately postoperatively were similar, but on D1, they were significantly higher in the ALI(-) group (p < 0.05). CONCLUSIONS: S100A12 increases in the early stage of development of ALI. sRAGE production increases in patients who do not develop ALI.

Effect of linezolid on cytokine production capacity and plasma endotoxin levels in response to lipopolysaccharide stimulation of whole blood.

The purpose of this study was to assess lipopolysaccharide (LPS)-stimulated cytokine production in the presence of linezolid (LZD) in comparison with the drug effect on the plasma endotoxin level. Peripheral venous whole-blood samples collected from five healthy subjects were stimulated with 10 microg/ml of LPS. LZD was then added to the LPS-stimulated blood samples at concentrations of 0, 2, 4, and 15 microg/ml , followed by incubation for 24 h at 37 degrees C in a 5% CO(2)-95% air atmosphere. Supernatants of the resultant cultures were assayed to determine the levels of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-10, monocyte chemoattractant protein (MCP)-1, and endotoxin. Significant decreases in the levels of TNF-alpha and IFN-gamma were observed in the LZD 2, 4, and 15 microg/ml groups as compared with that in the 0 microg/ml group (Dunnett's procedure; P < 0.05). The level of IL-10 tended to increase irrespective of the LZD concentration; however, no significant intergroup differences were observed [analysis of variance (ANOVA); P = 0.68]. No significant decrease of the endotoxin level was observed in the LZD 2, 4, or 15 microg/ml groups as compared with that in the 0 microg/ml group, with no significant intergroup differences (ANOVA; P = 0.83). No change in the MCP-1 levels was observed irrespective of the LZD concentration (ANOVA; P = 0.82). To conclude: (1) it appears possible that LZD inhibits the production of INF-gamma and TNF-alpha to a limited extent; (2) LZD did not exert any inhibitory effect on endotoxin production by bacteria, while suppressing cytokine production. The results indicate that LZD may have a significant role in saving the lives of patients with sepsis.

Laparoscopy-assisted major liver resections employing a hanging technique: the original procedure.

OBJECTIVE: To assess the feasibility, safety, and short-term outcomes of laparoscopy-assisted major liver resections. SUMMARY OF BACKGROUND DATA: The number of reports of laparoscopic major hepatectomies has gradually increased, and living donor hepatectomies for liver transplant have also recently been performed. However, because of the high degree of proficiency required, major hepatectomies have not been widespread. We developed an original procedure in which the liver is mobilized laparoscopically and resected by a hanging technique through a small incision. METHODS: Between November 2002 and December 2008, 43 patients underwent laparoscopy-assisted major liver resections (LAMLRs) in our institution for hepatocellular carcinoma, metastatic liver cancer, and benign diseases. RESULTS: LAMLRs were completed for 42 patients (97.7%). The median age was 62 years (range: 24-83 years). Preoperative diagnoses were hepatocellular carcinoma (n = 15), metastatic liver cancer (n = 19), and benign disease (n = 8). The types of liver resection consisted of the following: right trisectionectomy (n = 2), right hepatectomy (n = 14), left hepatectomy (n = 16), trisegmentectomy 4, 5, 8 (n = 2), right anterior sectionectomy (n = 4), and extended right posterior sectionectomy (n = 4). The median operating time was 317 minutes (range: 192-542 minutes) and median blood loss was 631 mL (range: 68-2785 mL). There were neither perioperative deaths nor reoperations. Five patients (11.9%) experienced postoperative complications, 2 patients (4.8%) showed bile leakage, and 3 patients (7.1%) developed wound infections. The median postoperative hospital stay was 13.5 days (range: 6-154 days). CONCLUSIONS: LAMLR with the hanging technique can be completed safely. The procedure can be performed by open liver surgeons; and thus may be widely performed in the future.

Mono-(2-ethylhexyl) phthalate targets glycogen debranching enzyme and affects glycogen metabolism in rat testis.

Phthalate esters are commonly used plasticizers; however, some are suspected to cause reproductive toxicity. Administration of high doses of di-(2-ethylhexyl) phthalate (DEHP) induces germ cell death in male rodents. Mono-(2-ethylhexyl) phthalate (MEHP), a hydrolyzed metabolite of DEHP, appears to be responsible for this testicular toxicity; however, the underlying mechanism of this chemical's action remains unknown. Here, using a one-step affinity purification procedure, we identified glycogen debranching enzyme (GDE) as a phthalate-binding protein. GDE has oligo-1,4-1,4-glucanotransferase and amylo-1,6-glucosidase activities, which are responsible for the complete degradation of glycogen to glucose. Our findings demonstrate that MEHP inhibits the activity of oligo-1,4-1,4-glucanotransferase, but not of amylo-1,6-glucosidase. Among various phthalate esters tested, MEHP specifically binds to and inhibits GDE. We also show that DEHP administration affects glycogen metabolism in rat testis. Thus, inhibition of GDE by MEHP may play a role in germ cell apoptosis in the testis.

[A case of metastatic lung and liver tumors from rectal cancer treated with oral UFT and CPT-11 by hepatic arterial infusion followed by FOLFOX and FOLFIRI].

The patient was a 62-year-old male who underwent a high anterior resection for rectal cancer with multiple liver metastases in June 2004. After the operation, 66 courses of weekly hepatic arterial infusion(HAI)therapy of 5-FU/Leucovorin( LV)were performed. Thereafter 14 courses of FOLFOX 4, 5 courses of FOLFIRI and 5 courses of FOLFOX 4 therapy were also sequentially performed. As a result of the CT examination, which revealed a new metastatic lesion in the liver and lung metastases, combination chemotherapy consisting of UFT and HAI of low-dose CPT-11 was administered in July 2007. After 1 cycle of this therapy, metastatic liver and lung tumors showed a reduction rate of 8.5% and 27.0%, respectively, without any adverse events. The elevated serum CEA (2,055 ng/mL)and CA19-9 (924 U/mL) levels decreased to 623 ng/mL and 332U /mL, respectively, after 1 cycle of the treatment. The combination of oral UFT and HAI of CPT-11 may therefore be a useful treatment for patients after FOLFOX and FOLFIRI therapy.

Portal vein reconstruction using a left renal vein graft for a patient with hilar cholangiocarcinoma.

We report a patient with hilar cholangiocarcinoma who underwent combined portal vein reconstruction using a left renal vein graft. A 68-year-old man was referred to the hospital with a one-week history of dark urine and jaundice. Cholangiography through the percutaneous transhepatic biliary drainage catheter and magnetic resonance cholangiopancreatography demonstrated complete obstruction of the hepatic primary confluence extended to the left secondary confluence. The patient underwent left hepatic lobectomy combined with total caudate lobectomy and extrahepatic bile duct resection. At operation, carcinoma invasion was observed from the portal trunk to the right portal branch. So, combined portal vein resection and graft interpose using left renal vein was performed. The caliber of left renal vein was wider than the right portal branch. No remarkable renal and hepatic dysfunction occurred postoperatively. In conclusion, left renal vein seems appropriate as an autograft when reconstructing the portal vein, especially main portal trunk, in patients with advanced hepatobiliary malignancies. It may be necessary to adjust the caliber when anastomosing the left renal vein to the right or left portal branch because the diameter of the left renal vein is usually wide.

Significance of extensive surgery including resection of the pancreas head for the treatment of gallbladder cancer--from the perspective of mode of lymph node involvement and surgical outcome.

The present study aimed to clarify the efficacy of extensive surgery, including pancreas head resection, for more complete lymphadenectomy in the treatment of gallbladder carcinoma. The study involved retrospective analyses of 65 consecutive patients with gallbladder carcinoma who underwent surgical resection between 1982 and 2003. Of these 65 patients, 41.5% displayed node-positive disease and among them 23.1% had positive para-aortic nodes. Of six node-positive 5-year survivors, five underwent pancreatoduodenectomy combined with S4aS5 hepatic subsegmentectomy. The 5-year survival rates were 76.2% for pN0, 30.0% for pN1, 45.8% for pN2, and 0% for pM1[lymph], respectively. Significant differences existed in survival rates. Postoperative recurrence was observed in 24.1% (13/54) of patients who underwent R0 resection. Of the four patients who displayed lymph node recurrence, two had pericholedocal and/or posterior pancreatoduodenal lymph node metastasis at the time of surgery and underwent pancreas-preserving regional lymphadenectomy. These results suggest that extensive resection, including resection of the pancreatic head, is effective in selected patients with up to pN2 lymph node metastasis, as long as complete removal of the cancer can be achieved. Pancreatoduodenectomy combined with S4aS5 hepatic subsegmentectomy should be considered when lymph node metastasis is obvious and the patient is in good condition.

[Endoscopic radiofrequency ablation for liver metastasis of colorectal cancer].

The application of radiofrequency ablation (RFA) for liver metastasis of colorectal cancer has not yet acquired an established status in clinical cancer therapy research. Removing as much tumor tissue as possible is desirable, but some cases do not allow optimal surgical ablation due to general condition of the patient and tumor status. We introduced endoscopic RFA for liver cancer in 2003, and have applied the procedure to 6 cases with H1 or H2 liver metastases of colorectal cancer to which surgical ablation could not be applied due to the poor general health of patients. Mean tumor diameter was 22.9 mm, and mean number of tumors per patient was 1.2. Tumor location was: S4, n = 2; S5, n = 1; S4, n = 1; S7, n = 2; and S8, n = 1. Mean frequency of session was 3.0. No complications occurred in any cases, and no reoperations were required. Although no recurrence of tumors in the vicinity of ablation was observed, 2 cases of each lung metastasis and intrahepatic recurrence were identified. Intrahepatic recurrence underwent hepatic arterial infusion (HAI) chemotherapy for simultaneous metastatic hepatic tumors (H2) prior to RFA, and relapses occurred in the metastatic focus where the efficacy of HAI was observed. At this point, 2 deaths were reported, 1 each from cancer and other diseases, and mean duration of survival after the procedure was 451.2 days. These results indicate that endoscopic RFA with good local control should be an available treatment for cases involving colorectal cancer with metastasis to the liver in which surgical ablation is difficult to apply.

Clinicopathological study of depth of subserosal invasion in patients with pT2 gallbladder carcinoma.

BACKGROUND: We examined whether depth of subserosal cancer invasion predicts lymph node involvement and survival in gallbladder carcinoma (GBC) patients with pathologicial subserosal invasion (pT2), to explore which patients benefit from radical second resection among patients with inapparent pT2 tumor. METHODS: Subjects comprised 31 patients with pT2 GBC. Thickness of the subserosal layer and vertical length of carcinoma invasion into the subserosa were measured under microscopy. Depth of subserosal invasion was divided subjectively into three categories: ss1, ss2, and ss3 (invasion of upper, middle, and lower thirds of the subserosal layer, respectively). Relationships between subserosal subclassification, histopathological factors, and prognosis were examined. RESULTS: Subserosal layers were significantly thicker (P < 0.001) in portions with cancer invasion (5.46 +/- 0.68 mm; range 1.0 approximately 13.75 mm) than those without cancer invasion (1.89 +/- 0.16 mm, range, 0.88 approximately 4.50 mm). Depth of carcinoma invasion into subserosa was 4.20 +/- 0.65 mm (range, 0.25 approximately 12.5 mm). Rate of lymphatic permeation, venous permeation, and lymph node involvement significantly increased with deeper subserosal invasion (P = 0.014, P = 0.027, P = 0.018, respectively). Among histopathological factors examined, only subserosal subclassification had a significant correlation with presence or absence of lymph node metastasis. Further, there was a significant correlation (P = 0.043) between the degree of subserosal invasion (ss1, ss2, and ss3) and involved nodal disease (pN0, pN1, and pM1 [lymph]). Although 5-year survival rates, according to the degree of subserosal invasion, tended to decrease with deeper invasion into the subserosal layer (ss1, 83.3%; ss2, 62.5%; ss3, 50.0%), no significant differences were noted. CONCLUSIONS: Pathological characteristics tend to become more aggressive with increasing depth of subserous carcinoma invasion in pT2 GBC. Depth of subserosal invasion is a predictor of presence and degree of lymph node metastasis in pT2 GBC. A sampling biopsy of the para-aortic nodes is recommended for inapparent pT2 GBC patients with subserosal invasion beyond one-thirds of the subserosal layer when they undergo radical second resection.

Cadmium exposure alters metabolomics of sulfur-containing amino acids in rat testes.

This study aimed to examine distribution of cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CSE), the hydrogen sulfide (H(2)S)-generating enzymes, and metabolomic alterations in sulfur-containing amino acids in rat testes exposed to stressors. Immunohistochemistry revealed distinct distribution of the two enzymes: CBS occurred mainly in Leydig cells and was also detectable in Sertoli cells and germ cells, whereas CSE was evident in Sertoli cells and immature germ cells involving spermatogonia. The amounts of CSE and CBS in testes did not alter in response to administration of cadmium chloride, an antispermatogenic stressor leading to apoptosis. Metabolome analyses assisted by liquid chromatography equipped with mass spectrometry revealed marked alterations in sulfur-containing amino acid metabolism: amounts of methionine and cysteine were significantly elevated concurrently with a decrease in the ratio between S-adenosylhomocysteine and Sadenosylmethionine, suggesting expansion of the remethylation cycle and acceleration of methyl donation. Despite a marked increase in cysteine, amounts of H(2)S were unchanged, leading to a remarkable decline of the H(2)S/cysteine ratio in the cadmium-treated rats. Under such circumstances, oxidized glutathione (GSSG) was significantly reduced, whereas reduced glutathione (GSH) was well maintained, and the GSH/GSSG ratio was consequently elevated. These results collectively showed that cadmium induces metabolomic remodeling of sulfur-containing amino acids even when the protein expression of CBS or CSE is not evident. Although detailed mechanisms for such a remodeling event remain unknown, our study suggests that metabolomic analyses serve as a powerful tool to pinpoint a critical enzymatic reaction that regulates metabolic systems as a whole.

[Oncologic emergencies associated with pancreatobiliary cancer].

Clinically common oncologic emergencies associated with pancreatobiliary cancer are gastrointestinal bleeding caused by duodenal invasion of pancreatic carcinoma, severe duodenal obstruction due to pancreatic carcinoma, and acute cholangitis accompanied by obstructive jaundice in patients with biliary tract carcinoma. When a patient with gallbladder cancer presents with acute cholecysitis, emergency surgery is sometimes performed on the basis of the latter diagnosis. Emergency procedures can also be required in the perioperative management of pancreatobiliary cancer, for example, in biliary peritonitis caused by detachment of a percutaneous transhepatic biliary drainage (PTBD) tube and in ruptured pseudoaneurysm due to postoperative pancreatic or biliary leakage. Nonsurgical procedures are usually initially selected for oncologic emergencies associated with pancreatobiliary cancer, because patients are likely to develop severe organ dysfunction and it is difficult to access directly and remove the pancreas or biliary tract during emergency surgery. When systemic conditions improve, it is necessary to evaluate the degree of disease progression and systemic conditions, and if feasible, the primary lesion should be surgically resected. When performing emergency cholecystectomy in patients with acute cholecystitis, thorough intraoperative investigation of resected specimens is important, considering the possibility of concomitant gallbladder carcinoma, since thorough examination cannot be performed in such emergency settings. Furthermore, when cholangitis accompanies pancreatobiliary cancer, emergency drainage should be considered as sepsis can develop rapidly.

Long-term results of central inferior (S4a+S5) hepatic subsegmentectomy and pancreatoduodenectomy combined with extended lymphadenectomy for gallbladder carcinoma with subserous or mild liver invasion (pT2-3) and nodal involvement: a preliminary report.

BACKGROUND/AIMS: Gallbladder carcinoma, especially advanced cancer that has invaded the subserosal or deeper layers, has a poor prognosis. Recently, radical operations combining resection of the liver and pancreas with extended lymph node dissection have been introduced to improve the prognosis of advanced gallbladder carcinoma. We have introduced central inferior (Couinaud's subsegments 4a and 5; S4a+S5) hepatic subsegmentectomy and pancreatoduodenectomy combined with extended lymphadenectomy for gallbladder carcinoma demonstrating subserous or mild liver invasion (pathological tumor stage pT2-3) and nodal involvement. METHODOLOGY: Morbidity, mortality, clinicopathological features, and long-term outcome were analyzed retrospectively for seven consecutive patients who underwent this radical operation. RESULTS: The postoperative morbidity rate was 57.1% and there was no surgical mortality. All patients had lymph node involvement: two had pN1 disease and five had pN2 disease. All patients underwent curative resection. Only one patient developed gallbladder carcinoma recurrence after resection and it involved the paraaortic lymph nodes. Five patients, three of whom displayed pN2 disease, survived longer than 5 years postoperatively with no evidence of disease. CONCLUSIONS: S4a+S5 hepatic subsegmentectomy and pancreatoduodenectomy combined with extended lymphadenectomy improve the long-term survival of gallbladder carcinoma with pT2-3 and nodal involvement. The presence of pN2 disease is not a contraindication for surgery. Further study is necessary to evaluate the usefulness of this radical procedure, especially as a standard operation.

Resection of liver metastasis from extrahepatic bile duct carcinoma previously treated by pancreatoduodenectomy.

No English report of a 5-year survivor with liver metastasis from periampullary carcinoma has been published to date. Here, we report a 52-year-old woman with liver metastasis previously treated with pylorus-preserving pancreatoduodenectomy for extrahepatic bile duct carcinoma, who underwent hepatectomy and has survived for more than 5 years without recurrence after the procedure. This outcome suggests that hepatic resection is effective for selected patients with liver metastasis from extrahepatic bile duct carcinoma previously treated by pancreatoduodenectomy. When a patient has a solitary metastasis of the liver with no other evidence of recurrence, surgical resection of the liver metastasis should be considered a valid option.