Preventing hypoxia/reoxygenation damage to hepatocytes by p66(shc) ablation: up-regulation of anti-oxidant and anti-apoptotic proteins

BACKGROUND/AIMS: Ischemia/reperfusion damage to the liver remains a serious concern in many clinical situations. Major mechanisms for this certainly include oxidative stress. METHODS: The effects of ablating the p66 isoform of ShcA (p66(shc)) on hypoxia/reoxygenation (H/R)-induced oxidative stress and cell injury in hepatocytes were investigated. RESULTS: Immediately after reoxygenation, AML12 cells were clearly under oxidative stress; many cells underwent apoptosis. However, knockdown of p66(shc) by specific RNAi markedly decreased cellular oxidative stress and H/R-induced apoptosis, as well as conferring resistance to H(2)O(2) insult. These data suggest that prevention of apoptosis conferred by ablation of p66(shc) results from changed ROS-scavenging, but not inhibition of ROS generation. These data were also confirmed in fibroblasts from p66(shc) knockout mice. Anti-oxidant molecules, such as MnSOD and Ref-1 and the anti-apoptotic molecule Bcl-xL were up-regulated, and pro-apoptotic FLICE was down-regulated, by ablation of p66(shc). Interestingly, catalase expression was not affected in p66(shc)-knockdown-AML12 cells although it is a major target in other cell types. CONCLUSIONS: Our findings suggest that in hepatocytes, ablation of p66(shc) is cytoprotective against H/R-induced oxidative stress, with MnSOD and Ref-1 playing critical roles, and with up-regulation of Bcl-xL and down-regulation of FLICE contributing jointly to preventing cells from undergoing oxidant-induced apoptosis.     

Antidepressant Properties of Aqueous Macerate from Gladiolus Dalenii Corms

Background

Gladiolus dalenii Van Geel (Iridaceae) has been used for the treatment of depression and psychotic disorders in African traditional medicine. The aim of this study was to assess the effect of the aqueous extract from the corm of Gladiolus dalenii and its possible mechanisms of action.

Materials and Methods

We assessed the antidepressant properties of G. dalenii corm aqueous extract in mice, using the open field, forced swimming, and tail suspension tests. Spontaneous locomotor activity of mice given various doses of G. dalenii extract (per os) was determined in the open field, whereas immobility was evaluated in the other two tests.

Results

Extract maximal effect was observed at 15 mg/kg, as mice displayed a marked reduction in immobility time in both the forced swimming test (80%) and the tail suspension test (66%). In further studies aimed at investigating the mechanism of action of G. dalenii extract, the latter significantly antagonized the effect of N-methyl-D-aspartate (NMDA, 75 mg/kg) at both the doses 15 mg/kg (p<0.001) and 150 mg/kg (p=0.004). A significant reduction in immobility time was also observed following treatment with combinations of a sub-effective dose of extract (7.5 mg/kg) with either the NMDA receptor antagonist D-(−)-2-amino-7-phosphonoheptanoic acid (D-AP7, 50 mg/kg, P< 0.001), the serotonin reuptake inhibitor fluoxetine (5 and 10 mg/kg, P< 0.001and P< 0.001 respectively), and the multi-target antidepressant imipramine (5 and 10 mg/kg, P< 0.001 and P< 0.001 respectively). Moreover, neither G. dalenii extract alone nor its combinations with NMDA ligands imipramine and fluoxetine enhanced mouse spontaneous locomotor activity.

Conclusion

Altogether, these results suggest that G. dalenii has antidepressant properties, probably mediated through interactions with NMDA, serotonin and/ or noradrenergic systems, and may justify its use in traditional medicine.     

Mechanical environment of the supraspinatus tendon: a two-dimensional finite element model analysis

We performed 2-dimensional finite element model analysis to estimate the mechanical environment of the supraspinatus tendon. The geometric shape of the finite element model was determined by magnetic resonance imaging of a normal human shoulder obtained at 0 degrees, 30 degrees, and 60 degrees of abduction, whereas the histologic location of noncalcified and calcified fibrocartilage was determined from a cadaveric specimen. The supraspinatus tendon was pulled proximally with the force of 10 N at 0 degrees, 53 N at 30 degrees, and 115 N at 60 degrees of abduction. The area of high principal stress maximum was observed on the articular side of the supraspinatus tendon, which shifted toward the insertion as the arm was abducted. High stress concentration on the articular side of the supraspinatus tendon near its insertion during arm elevation may explain the frequent occurrence of rotator cuff tears at this site.     

A case of hepatic portal venous gas caused by chemo-radiation therapy for an advanced esophageal cancer

Hepatic portal venous gas(HPVG)is a rare condition with a poor prognosis. A 40-year-old man underwent esophagectomy for stage IV esophageal cancer followed by chemotherapy. Four months later, he admitted to our hospital because of the increases of residual tumors and started chemoradiotherapy(CRT)with 5-FU, CDDP and radiation. Computed tomography(CT)scan revealed PR, and blood examination showed decreases in WBC and platelet counts. Fourty days after CRT, he suddenly complained severe pain in the left chest and abdomen, and vomiting. CT scan showed HPVG in the left lobe of the liver and pneumatosis cystoides intestinalis in the wall of the gastric tube. He died of multiple organ failure. To our knowledge, this is a first case of HPVG associated with CRT for esophageal cancer.     

Hydrogen Gas Ameliorates Hepatic Reperfusion Injury After Prolonged Cold Preservation in Isolated Perfused Rat Liver

Hydrogen gas reduces ischemia and reperfusion injury (IRI) in the liver and other organs. However, the precise mechanism remains elusive. We investigated whether hydrogen gas ameliorated hepatic I/R injury after cold preservation. Rat liver was subjected to 48‐h cold storage in University of Wisconsin solution. The graft was reperfused with oxygenated buffer with or without hydrogen at 37° for 90 min on an isolated perfusion apparatus, comprising the H₂(+) and H₂(?) groups, respectively. In the control group (CT), grafts were reperfused immediately without preservation. Graft function, injury, and circulatory status were assessed throughout the perfusion. Tissue samples at the end of perfusion were collected to determine histopathology, oxidative stress, and apoptosis. In the H₂(?) group, IRI was indicated by a higher aspartate aminotransferase (AST), alanine aminotransferase (ALT) leakage, portal resistance, 8‐hydroxy‐2‐deoxyguanosine‐positive cell rate, apoptotic index, and endothelial endothelin‐1 expression, together with reduced bile production, oxygen consumption, and GSH/GSSG ratio (vs. CT). In the H₂(+) group, these harmful changes were significantly suppressed [vs. H₂(?)]. Hydrogen gas reduced hepatic reperfusion injury after prolonged cold preservation via the maintenance of portal flow, by protecting mitochondrial function during the early phase of reperfusion, and via the suppression of oxidative stress and inflammatory cascades thereafter.     

Comparison of timing and decision-makers of do-not-resuscitate orders between thoracic cancer and non-cancer respiratory disease patients dying in a Japanese acute care hospital

Purpose

The aim of the study was to compare timing and decision-makers of do-not-resuscitate (DNR) orders between patients with end-stage thoracic cancer and non-cancer respiratory diseases in a Japanese acute care hospital.

Methods

This study retrospectively reviewed the medical records of patients who died between January 2008 and March 2013 in the Department of Respiratory Medicine of Osaka Police Hospital, a teaching and acute care hospital. We compared the decision-making process, especially timing and decision-maker, of DNR orders between patients with thoracic cancer and patients with non-cancer respiratory diseases.

Results

There were 300 cancer patients and 147 non-cancer patients. Cancer patients were significantly younger, were hospitalized more frequently and for longer, were more likely to have a DNR order placed earlier and decided in advance of last admission, and were more likely to have normal cognitive function at the time of the DNR order than non-cancer patients. Spouses of cancer patients were more likely to participate in DNR discussion. Only approximately 6 % of patients participated in DNR discussion in both groups. Cancer patients less frequently received aggressive treatment at the end of life (EOL) and were more likely to die in general wards than in intensive care units.

Conclusions

Our study found that most Japanese patients, with or without cancer, who died in an acute care respiratory department, were not included in DNR discussions and that familial surrogates usually made the DNR decision at the EOL.     

The initiation activity of norfloxacin does not result in the induction of hepatocellular tumors in rats

In order to examine whether the in vivo initiation activity of a new quinolone antimicrobial agent -- norfloxacin (NFLX) -- results in the induction of hepatocellular tumors, F344 male rats were subjected to two-thirds partial hepatectomy and oral administration of 1500 or 750mg/kg BW of NFLX or the vehicle once daily for 3 weeks. From 2 weeks after the completion of NFLX treatment, the rats were given 500ppm phenobarbital (PB) in their drinking water for 51 weeks. After the promotion treatment with PB for 17, 34, or 51 weeks, the rats were euthanized under ether anesthesia, and the NFLX-induced hepatic tumors were examined macroscopically by thoroughly sectioning the liver at 5-mm intervals. The liver slices, one each from all liver lobes, were fixed in 10% neutral buffered formalin for immunohistochemical examination of glutathione S-transferase placental form (GST-P) positive foci. NFLX increased neither the incidence of macroscopic hepatic tumors nor the mean number or area of GST-P positive foci. These results suggest that under the present study conditions, the initiation activity of NFLX does not result in the induction of hepatocellular tumors in rats; thus, the initiation activity of NFLX is extremely weak.     

Probiotic effects of orally administered Lactobacillus salivarius WB21-containing tablets on periodontopathic bacteria: a double-blinded, placebo-controlled, randomized clinical trial

Aim: This study was designed to evaluate whether the oral administration of lactobacilli could change the bacterial population in supra/subgingival plaque.
Material and Methods: Sixty-six healthy volunteers without severe periodontitis were randomized into two groups to receive lactobacilli or placebo for 8 weeks (8W): the test group ( n =34) received 2.01 × 10⁹ CFU/day of Lactobacillus salivarius WB21 and xylitol in tablets; the control group ( n =32) received placebo with xylitol. Supra/subgingival plaque samples were collected at the baseline and after 4 weeks (4W) and 8W. The bacterial amounts in plaque samples were analysed by quantitative real-time polymerase chain reaction.
Results: The numerical sum of five selected periodontopathic bacteria in the test group was decreased significantly in subgingival plaque at 4W [odds ratio (OR)=3.13, 95% confidence intervals (CI)=1.28–7.65, p =0.012]. Multivariate analysis showed that significantly higher odds were obtained for the reduction of Tannerella forsythia in subgingival plaque of the test group at both 4W (OR=6.69, 95% CI=2.51–17.9, p <0.001) and 8W (OR=3.67, 95% CI=1.45–9.26, p =0.006).
Conclusion: Oral administration of probiotic lactobacilli reduced the numerical sum of five selected periodontopathic bacteria and could contribute to the beneficial effects on periodontal conditions.     

Within-Day Energy Balance and Metabolic Suppression in Male Collegiate Soccer Players

Metabolic suppression due to relative energy deficiency can cause various physiological impairments in athletes. The purpose of this study was to evaluate within-day energy balance (WDEB) and the ratio between measured and predicted resting energy expenditure (REE_ratio) and to investigate the relationships between the markers of metabolic suppression. Ten male collegiate soccer players completed a 7-day food diary, physical activity, and heart rate records during the training and rest days. Energy intake (EI) and energy expenditure (EE) were analyzed to evaluate WDEB components. Body composition was measured using dual-energy X-ray absorptiometry (DXA), and blood sampling was conducted for hormonal analysis. The REE was measured using the Douglas bag method and predicted using the DXA-predicted method to calculate the REE_ratio. Participants were categorized into the normal (REE_ratio ≥ 0.94, n = 5) and suppressed (REE_ratio < 0.94, n = 5) groups. There were no group differences in the components of WDEB, except diet-induced thermogenesis (DIT), but EI was significantly higher in the normal group than in the suppressed group (7-day total: 3660 ± 347 vs. 3024 ± 491 kcal/day, p = 0.046 and rest days: 3772 ± 463 vs. 2796 ± 800 kcal/day, p = 0.046). Analysis of hormonal markers of metabolic suppression only showed a significant positive association between insulin-like growth factor-1 (IGF-1) and REE_ratio (r = 0.771, p = 0.009). The relationships between metabolic suppression and the markers of energy deficiency were inconclusive. There are possible associations of insufficient EI and IGF-1 levels with metabolic suppression, and further study is required to understand energy deficiency in male soccer players.     

A case of Stage IV gastric cancer with liver and peritoneal metastases responding completely to tailored S-1/CPT- 11 combination therapy

A 75-year-old man with advanced gastric cancer underwent distal gastrectomy with lymph node dissection(D1)and Roux-en Y reconstruction. Pathological staging was Stage IV (T3N3P1CY1M1), and curability was Cur C. He started adjuvant chemotherapy with oral administration of S-1(100 mg/body weight), but experienced grade 3 anorexia for one month. Abdominal computed tomography(CT)2 months postoperatively showed multiple liver metastases and ascites. We then conducted tailored S-1/CPT-11 as second-line chemotherapy(S-1 80 mg/body weight on days 1-5 and 8-12, CPT-11 60 mg/body weight on days 1 and 8). After 5 courses of this therapy, CT showed that the liver metastases and ascites had disappeared, leading to a complete response(CR). The only adverse event was general grade 1 fatigue. He continues to undergo oral administration of S-1(80 mg/body weight)as maintenance therapy, and maintained CR for 12 months since undergoing chemotherapy. Adverse events in tailored S-1/CPT-11 combination therapy are mild and tolerable, making this regimen a potential therapeutic strategy for patients with advanced or recurrent gastric cancer.