Development of pyrrole-imidazole polyamide for specific regulation of human aurora kinase-A and -B gene expression.

Pyrrole-imidazole polyamide (PIP) is a nuclease-resistant novel compound that inhibits gene expression through binding to the minor groove of DNA. Human aurora kinase-A (AURKA) and -B (AURKB) are important regulators in mitosis during the cell cycle. In this study, two specific PIPs (PIP-A and PIP-B) targeting AURKA and AURKB promoter regions were designed and synthesized, and their biological effects were investigated by several in vitro assays. PIP-A and PIP-B significantly inhibited the promoter activities, mRNA expression, and protein levels of AURKA and AURKB, respectively, in a concentration-dependent manner. Moreover, 1:1 combination treatment with both PIPs demonstrated prominent antiproliferative synergy (CI value [ED(50)] = 0.256) to HeLa cells as a result of inducing apoptosis-mediated severe catastrophe of cell-cycle progression. The novel synthesized PIP-A and PIP-B are potent and specific gene-silencing agents for AURKA and AURKB.

Prolonged mild therapeutic hypothermia versus fever control with tight hemodynamic monitoring and slow rewarming in patients with severe traumatic brain injury: a randomized controlled trial.

Although mild therapeutic hypothermia is an effective neuroprotective strategy for cardiac arrest/resuscitated patients, and asphyxic newborns, recent randomized controlled trials (RCTs) have equally shown good neurological outcome between targeted temperature management at 33 °C versus 36 °C, and have not shown consistent benefits in patients with traumatic brain injury (TBI). We aimed to determine the effect of therapeutic hypothermia, while avoiding some limitations of earlier studies, which included patient selection based on Glasgow coma scale (GCS), delayed initiation of cooling, short duration of cooling, inter-center variation in patient care, and relatively rapid rewarming. We conducted a multicenter RCT in patients with severe TBI (GCS 4-8). Patients were randomly assigned (2:1 allocation ratio) to either therapeutic hypothermia (32-34 °C, n = 98) or fever control (35.5-37 °C, n = 50). Patients with therapeutic hypothermia were cooled as soon as possible for ≥ 72 h and rewarmed at a rate of <1 °C/day. All patients received tight hemodynamic monitoring under intensive neurological care. The Glasgow Outcome Scale was assessed at 6 months by physicians who were blinded to the treatment allocation. The overall rates of poor neurological outcomes were 53% and 48% in the therapeutic hypothermia and fever control groups, respectively. There were no significant differences in the likelihood of poor neurological outcome (relative risk [RR] 1.24, 95% confidence interval [CI] 0.62-2.48, p = 0.597) or mortality (RR 1.82, 95% CI 0.82-4.03, p = 0.180) between the two groups. We concluded that tight hemodynamic management and slow rewarming, together with prolonged therapeutic hypothermia (32-34 °C) for severe TBI, did not improve the neurological outcomes or risk of mortality compared with strict temperature control (35.5-37 °C).

Effect of posttraumatic hyperglycemia on contusion volume and neutrophil accumulation after moderate fluid-percussion brain injury in rats.

The purpose of this study was to evaluate the effects of posttraumatic hyperglycemia on contusion volume and neutrophil accumulation following moderate traumatic brain injury (TBI) in rats. A parasagittal fluid-percussion (F-P) brain injury (1.8-2.1 atm) was induced in male Sprague-Dawley rats. Rats were then randomized into four trauma groups (n = 7/group) by the timing of dextrose injection (2.0 gm/kg/ip), which included (1) early (E) group: 5 min after TBI; (2) delayed (D) group: 4 h after TBI; (3) 24-h group: 24 h after TBI; or (4) control (C) group: no dextrose injection. A sham operated control group also received dextrose to document physiological parameters (n = 4). Rats were perfusion fixed 3 days following TBI, and the brains were processed for routine histopathological and immunocytochemical analysis. Contusion areas and volumes, as well as the frequency of myeloperoxidase immunoreactive polymorphonuclear leukocytes (PMNLs) were determined. Dextrose injections significantly increased blood glucose levels (p < 0.005) in all treated groups. Although acute hyperglycemia following TBI did not significantly affect total contusion volume, contusion area was significantly elevated in the early treatment group. In addition, early posttraumatic hyperglycemia enhanced neutrophil accumulation in the area of the cortical contusion (p < 0.005). In contrast, delayed induced hyperglycemia (i.e., 4 h, 24 h) did not significantly affect histopathological outcome or neutrophil accumulation. Taken together, these findings indicate that acute but not delayed hyperglycemia aggravates histopathological outcome and increased accumulation of PMNLs. Posttraumatic hyperglycemia in the acute phase may worsen traumatic outcome by enhancing secondary injury processes, including inflammation.

Risk factors for intraoperative hypotension in traumatic intracranial hematoma.

Patients suffering from traumatic intracranial hemorrhage (TICH) may experience an episode of catastrophic intraoperative hypotension (IHT), after decompression of the brain. The aim of this study was to investigate the risk factors for IHT during emergency craniotomy A total of 67 patients, who underwent emergency craniotomy due to TICH, were divided into two groups: IHT ( n=31 ) or without IHT ( n=36 ). Data concerning (1) age; (2) gender; (3) mechanism of injury; (4) Glasgow Coma Scale (GCS) on admission; (5) abnormality of the pupils (anisocoria or mydriasis); (6) mean arterial blood pressure; (7) heart rate; (8) time elapsed before craniotomy from injury; (9) initial brain CT scans; (10) duration of craniotomy; and (11) total infusion or urine volume until craniotomy were collected prospectively as IHT risk factors. Low GCS score (<5), tachycardia (heart rate >112min(-1)) and hypertension (mean blood pressure >131mmHg) before emergency craniotomy were strongly ( P<0.05 ) associated with IHT. Delayed surgery (>173min until craniotomy) also had a significant ( P<0.005 ) effect on IHT. The risk factors for IHT were considered as a low GCS score on admission, tachycardia, hypertension before emergency craniotomy and delayed surgery. These results suggested the patients with IHT had a high sympathetic tone before emergency craniotomy A sudden reduction in sympathetic tone after surgical decompression of the brain might cause IHT. We concluded that an important factor in the occurrence of IHT was not only the injury severity, but also the balance between sympathetic and parasympathetic activity before decompression surgery.

The peculiar processing of glucagon and glucagon-related peptides in patients after pancreatectomy.

BACKGROUND/AIMS: Although, glucagon has been detected even in the serum of totally pancreatectomized patients and the origin was suggested to be the intestine, the kinetics of glucagon is not well known after pancreatectomy. In this study, we investigated the kinetics of glucagon and glucagon-related peptides and discuss the glucagon processes in the pancreas and intestine in pancreatectomy patients. METHODOLOGY: Seven patients who had undergone pancreatoduodenectomy reconstruction using Child's procedures (distal pancreatojejunostomy, choledochojejunostomy and gastrojejunostomy) (group PD) and five patients who had undergone distal pancreatectomy (group DP) served as the subjects of this study. In addition to these two groups, five patients who had undergone gastrectomy reconstructions using the Billroth II procedure (group GX), to examine whether the alimentary tract reconstructions themselves would have any effect on the kinetics of glucagon, and 10 normal subjects (group C) were also enrolled in this study. All patients received a 75-g oral glucose tolerance test in the early morning fasting state. Serum glucagon levels were assessed using the glucagon non-specific N-terminal (glucagon-like immunoreactivity: GLI) and specific C-terminal (immunoreactive glucagon: IRG) radioimmunoassays. The molecular forms of these glucagon-related peptides were also estimated using the gel filtration chromatography method before and after the 75-g oral glucose load. RESULTS: After the glucose load, serum GLIs were increased significantly in groups GX and PD suggesting that these were affected by the alimentary tract reconstructions. Serum IRGs including true pancreatic glucagon were slightly increased in groups PD and DP after oral glucose load suggesting that these paradoxical responses might be associated with the glucose tolerance deficiencies observed in both groups, but not associated with the alimentary tract reconstruction. CONCLUSIONS: The paradoxical rise of IRGs based on the findings of gel filtration chromatography were possibly due to the generated peculiar glicentin-like peptide and pancreatic glucagon from the glucagon precursor, preproglucagon, after pancreatectomy, which is processed in association with the glucose tolerance deficiencies after pancreatectomy.

Peculiar glucagon processing in the intestine is the genesis of the paradoxical rise of serum pancreatic glucagon in patients after total pancreatectomy.

BACKGROUND/AIMS: Although glucagon has been detected even in the serum of totally pancreatectomized patients and the origin was suggested to be the intestine, the kinetics of glucagon are not well known after pancreatectomy. In the present study, we investigated the kinetics of glucagon and glucagon-related peptides in pancreatectomy patients and discuss the glucagon processes. METHODOLOGY: Ten patients who had undergone total pancreatoduodenectomy reconstruction using Billroth II type procedures (group PX) and 12 normal subjects (group C) were also enrolled in this study. All patients received a 75-g oral glucose tolerance test in the early morning fasting state. Serum glucagon levels were assessed using the glucagon specific C-terminal (immunoreactive glucagon: IRG) and nonspecific N-terminal (glucagon-like immunoreactivity: GLI) radioimmunoassays. The molecular forms of these glucagon-related peptides were also estimated using the gel filtration chromatography method before and after the oral glucose load. RESULTS: After the glucose load, serum GLIs were increased significantly in group PX suggesting that these were affected by the alimentary tract reconstructions. Serum IRGs were significantly increased in group PX, but decreased in group C after oral glucose load suggesting that these paradoxical increased responses in group PX might be associated with the insulin secretion deficiencies, but not associated with the alimentary tract reconstruction. CONCLUSIONS: The paradoxical rise in IRGs based on the findings of gel filtration chromatography in group PX were possibly due to the generated peculiar glicentin-like peptide from the glucagon precursor, preproglucagon, after total pancreatectomy, which might be processed in intestines in association with the insulin deficiencies.

Metabolic modulation by concomitant administration of insulin and glucagon in pancreatectomy patients.

BACKGROUND/AIMS: The metabolic kinetics after a total pancreatectomy, especially the postoperative loss of pancreatic endocrine function including insulin and glucagon secretions, remain unexplored. We examined the metabolic status of patients after a total pancreatectomy under insulin control and the effects of glucagon administration on metabolic kinetics. METHODOLOGY: The metabolic assessments of 11 patients who had undergone total pancreatectomy were performed using indirect calorimetry with blood amino acid and lipid analysis in the following three metabolic conditions: 1) under conventional insulin control alone, 2) when a pharmacologically active amount of glucagon was administered, and 3) when an additional amount of insulin was administered concomitantly to attain the insulin/glucagon molar ratio in the serum similar to the physiological value. RESULTS: Under conventional insulin control alone, the level of calorigenesis from proteins was decreased mainly due to an impairment in the utilization of glycogenic amino acids. Furthermore, carbohydrates were not sufficiently utilized, and lipid metabolism was increased as a result of the reduced lipid catabolism. When a pharmacologically active amount of glucagon was administered, the utilization of glycogenic amino acids and lipids increased. However, the utilization of carbohydrates was slightly reduced. When an additional amount of insulin was administered, an increased utilization of protein was observed. At the same time, marked improvement was observed in the utilization of carbohydrates which was reduced in 1) and 2) indicated above. Furthermore, the utilization of lipids in calorigenesis was not reduced, but it was reduced in lipid catabolism. CONCLUSIONS: The above findings indicate the importance of administering insulin and glucagon concomitantly and maintaining the blood insulin/glucagon ratio at a level similar to the physiological value regardless of the administered dosages of these two hormones in order to improve the energy and nutritional metabolism following a total pancreatectomy.

Moderate low temperature preserves the stemness of neural stem cells and suppresses apoptosis of the cells via activation of the cold-inducible RNA binding protein.

We hypothesized that one of the mechanisms underlying the protection of brain injury by therapeutic hypothermia is associated with preservation of neural stem cells. We investigated effects of moderate low temperature and the contribution of a cold-inducible molecule for the stemness of neural stem cells. The MEB5 mouse neural stem cell line was cultured in the presence or absence of EGF, and apoptosis, mRNA expression, and immunocytochemistry of the differentiation markers nestin and GFAP were evaluated at 37 or 32°C. We investigated the contribution of the cold-inducible RNA binding protein (CIRP) on apoptosis and differentiation of MEB5 cells at 32°C. EGF deprivation increased the number of apoptotic cells, decreased expression of nestin, and increased expression of GFAP. The moderate low temperature prevented apoptosis and decreases in expression of GFAP in MEB5 by EGF deprivation. The moderate low temperature significantly increased expression of CIRP. siRNA against CIRP significantly increased the apoptotic cell population of MEB5 cells via EGF deprivation at 32°C. These findings suggest that moderate low temperature preserved stemness of neural stem cells and prevented cell apoptosis via the stimulation of CIRP, and one of the mechanisms of rescue of brain injury by the moderate hypothermia is associated with preservation of neural stem cells.

Management of pitfalls for the successful clinical use of hypothermia treatment.

Therapeutic hypothermia is a promising method for controlling intracranial pressure (ICP) in severely brain-injured patients. However, clinical data regarding the effect of brain hypothermia on overall outcome of these patients is limited. This may be because there are specific pitfalls associated with the clinical management of induced hypothermia in patients with severe traumatic brain injury (TBI). These pitfalls may be avoided by preventing specific risk factors when cooling is induced and with rewarming. However, these risk factors have not been well systematically discussed in the literature. In this paper, three categories of clinical issues regarding the management of brain hypothermia are discussed: (1) stress-induced secondary brain injury mechanisms; (2) technical aspects of intensive care unit (ICU) cooling management; and (3) rewarming rates and methods. For patients with a Glasgow Coma Scale (GCS) score of less than 8, management of stress-induced insulin-resistant hyperglycemia, and unstable systemic circulation due to impaired cardiac contractility are especially important. For example, in our experience, posttraumatic hyperglycemia, exacerbated by cooling, may be ameliorated by the administration of a ketone body with mannitol. Prevention of selective free radical damage to neurons is also an important target for successful brain hypothermia treatment. Taken together, it is clear that several orchestrated steps should be initiated to enhance the protective effects of hypothermia therapy and prevent these possible pitfalls.

Resuscitative value of B-type natriuretic peptide in comatose survivors treated with hypothermia after out-of-hospital cardiac arrest due to cardiac causes.

BACKGROUND: Two randomized studies have shown a neurological benefit of therapeutic hypothermia in comatose survivors after out-of-hospital cardiac arrest, but there are no studies of the cardiac neurohormone of B-type natriuretic peptide (BNP) in patients treated with hypothermia. METHODS AND RESULTS: A prospective study was conducted of 109 comatose patients who were treated with mild hypothermia after out-of-hospital sudden cardiac arrest due to cardiac causes and whose BNP level was measured on arrival at the emergency room. The primary endpoint was a favorable neurological outcome at the time of hospital discharge. A total of 45 of the 109 patients had a favorable neurological outcome. The unadjusted rate of a favorable neurological outcome decreased in a stepwise fashion among patients in increasing quartiles of BNP level (p<0.001) and this association remained significant in subgroups of patients. The BNP cutoff value of 80 pg/ml for a favorable neurological outcome had an accuracy of 87.2%. In the multiple logistic-regression analysis, a BNP level of 80 pg/ml or less was an independent predictor of favorable neurological outcome. CONCLUSIONS: The measurement of BNP was found to provide valuable information regarding the neurological outcome of comatose survivors treated with mild hypothermia after out-of-hospital cardiac arrest due to cardiac causes.

Prognostic value of blood glucose in patients with cardiogenic shock.

BACKGROUND: Although an elevated blood glucose has prognostic value in cardiovascular disease, few data are available regarding its prognostic value for patients across the spectrum of cardiogenic shock. METHOD AND RESULTS: A total of 81 patients with cardiogenic shock whose blood glucose and adrenaline were measured on arrival at the emergency room (ER) were enrolled in this prospective study. The primary endpoint was death from any cause in hospital. The rate of death was 12.3% (10/81), and the glucose level was lower among patients who were discharged alive than among those who died (8.7+/-3.7 mmol/L vs 13.8+/-6.7 mmol/L, p<0.001). The unadjusted rate of death increased in a stepwise fashion among patients in increasing quartiles of glucose level (p<0.05). The blood glucose level of 9.2 mmol/L had the highest combined sensitivity and specificity for the identification of death. In the multiple logistic-regression analysis for the primary outcome, the adjusted odd ratio for a glucose level of 9.2 mmol/L or more was 5.8 (95% confidence interval, 1.0-32.8, p=0.047). There was a significant positive correlation between the glucose and adrenaline levels (R=0.726, p<0.0001). CONCLUSION: The measurement of blood glucose level on ER arrival provides predictive information for use in risk stratification across the spectrum of cardiac emergencies complicated by cardiogenic shock.

B-type natriuretic peptide as a marker of resuscitation in patients with cardiac arrest outside the hospital.

BACKGROUND: Although the circulating concentration of B-type natriuretic peptide (BNP) has both a prognostic and diagnostic value in heart disease, no data are available regarding its resuscitative value for out-of-hospital cardiac arrest. METHODS AND RESULTS: The present study was a prospective study of 401 patients whose BNP was measured on arrival at the emergency room after an out-of-hospital cardiac arrest with a cardiac cause. The primary endpoint was survival to hospital discharge. The unadjusted rate of survival to hospital discharge decreased in a stepwise fashion among patients in increasing quartiles of BNP concentration (p<0.001). After adjusting for independent predictors of resuscitation, the odds ratios for survival to hospital discharge in the second, third and fourth quartiles of BNP were 0.13 (95% confidence interval (CI), 0.04-0.46), 0.10 (95% CI, 0.03-0.41), and 0.004 (95% CI, 0.00-0.16), respectively. The BNP cutoff value of 100 pg/ml for survival had a sensitivity of 83% and a negative predictive value of 96%. CONCLUSIONS: The measurement of BNP was found to provide valuable predictive information for survival to hospital discharge in patients with out-of-hospital cardiac arrest of cardiac etiology.

An early and complete reperfusion strategy for acute myocardial infarction using fibrinolysis and subsequent transluminal therapy--The FAST trial.

The efficacy and safety of fibrinolysis and subsequent transluminal (FAST) therapy were evaluated in 195 patients with acute myocardial infarction (AMI) for the early achievement of thrombolysis-in-myocardial-infarction grade 3 (TIMI-3) flow in the infarct-related artery. Intravenous thrombolysis using the optimal dose of a thrombolytic agent was initiated immediately after arrival in the emergency room, followed by coronary angiography and adjuvant percutaneous coronary intervention. A comparison of the thrombolysis alone (n=83) and thrombolysis plus intervention (n=112) groups showed significant differences in the time interval from hospital arrival to achievement of TIMI-3 flow (66.2+/-23.7 vs 111.6+/-29.6 min, p<0.0001), creatine kinase-MB release (295+/-201 vs 468+/-322 U/L, p=0.0003) and peak troponin T (23.6+/-16.9 vs 38.9+/-25.9 ng/ml, p<0.0001). No significant differences were observed in either 30-day mortality or complications. The TIMI-3 flow at the initial angiography was significantly higher with a single bolus of mutant tissue-type plasminogen activator (t-PA) monteplase than with an accelerated infusion of t-PA (60% vs 32%, p=0.005). In conclusion, the early restoration of TIMI-3 flow by FAST therapy reduced the degree of myocardial damage with a low risk of complications. TIMI-3 flow was achieved at an earlier stage with monteplase and this agent may be beneficial in the FAST therapy.