Aberrant DNA methylation of alternative promoter of DLC1 isoform 1 in meningiomas.

DLC1 encodes GTPase-activating protein with a well-documented tumor suppressor activity. This gene is downregulated in various tumors through aberrant promoter hypermethylation. Five different DLC1 isoforms can be transcribed from alternative promoters. Tumor-related DNA methylation of the DLC1 isoform 1 alternative promoter was identified as being hypermethylated in meningiomas in genome-wide DNA methylation profiling. We determined the methylation pattern of this region in 50 meningioma FFPE samples and sections of 6 normal meninges, with targeted bisulfite sequencing. All histopathological subtypes of meningiomas showed similar and significant increase of DNA methylation levels. High DNA methylation was associated with lack of DLC1 protein expression in meningiomas as determined by immunohistochemistry. mRNA expression levels of 5 isoforms of DLC1 transcript were measured in an additional series of meningiomas and normal meninges. The DLC1 isoform 1 was found as the most expressed in normal control tissue and was significantly downregulated in meningiomas. Transfection of KT21 meningioma cell line with shRNA targeting DLC1 isoform 1 resulted in increased activation of RHO-GTPases assessed with pull-down assay, enhanced cell migration observed in scratch assay as well as slight increase of cell metabolism determind by MTT test. Results indicate that isoform 1 represents the main pool of DLC1 protein in meninges and its downregulation in meningiomas is associated with hypermethylation of CpG dinucleotides within the corresponding promoter region. This isoform is functional GAP protein and tumor suppressor and targeting of its expression results in the increase of DLC1 related cell processes: RHO activation and cell migration.

[Hygiene status of ambulances and equipment in rescue services]. / Hygienestatus von Fahrzeugen und Ausrüstung im Rettungsdienst.

PURPOSE: Focus on hygienic management in ambulances is presented based on findings from hygienic microbiological sampling. METHODS: In 44 ambulances, the handwashing area, equipment for artificial respiration, insufflation, intubation, aspiration, intravasal catheterisation, blood-pressure measurement, and sterile materials storage were examined hygienically and microbiologically using the following methods: water sampling, imprint technique, and swab technique with subsequent cultivation on blood-, endo-, and Sabouraud agar. RESULTS: The highest contamination was found in the hand-washing area and on insufflation equipment (up to > or = 100,000 cfu/ml), where proof of the potential pathogens Pseudomonas, Acinetobacter, and Alcaligenes spp. was found. The second highest contamination level was found on the sphygmomanometer cuffs, stethoscopes, and respirator masks (e.g., Enterococci and S. aureus were identified). Apparently, the germs chiefly originate from the drinking water at the handwashing station, from the environment and mucous membranes from the skin, and intestines of the ambulance personnel. The predominant hygienic deficiensies were found in the equipment of the handwashing areas, and in the storage of resuscitation equipment and sterile materials. CONCLUSIONS: Based on the results of hygienic microbiological analysis, the main focus of hygienic measures--besides hand disinfection--should be on spray disinfection of the sphygmomanometer cuffs and stethoscopes as well as disinfection of ambulance equipment and storage areas for sterile devices, supplemented by dust protection of ready-to-use materials and devices. Stationary dispensers for hand disinfection possessing at least midlength levers must become the standard in ambulances, and conventional handwashing basins with plastic drinking-water containers and pump must be eliminated entirely.

In anterior myocardial infarction, frequency domain is better than time domain analysis of the signal-averaged ECG for identifying patients at risk for sustained ventricular tachycardia.

Low amplitude signals at the end of the QRS in patients with prior myocardial infarction (MI) are related to fragmentation of the electrical impulse in ventricular myocardium and are known to correlate with an increased risk of sustained ventricular tachycardia (VT). We hypothesized that in patients with anterior MI (AMI), earlier activation of the damaged anterior wall would cause an earlier fragmentation of the signal-averaged ECG (SAECG) signal, making conventional time domain analysis of late potentials difficult. We performed SAECG in 213 patients (62 with AMI and 58 with inferior MI [IMI]). Fifty-seven had prior sustained VT; 23 with AMI and 24 with IMI. We examined the standard time domain SAECG parameters including the duration of the filtered QRS (40-250 Hz), the duration of the late QRS < 40 microV, and the root mean square amplitude of the last 40 msec of the QRS. We also examined the power law scaling (PLS) in the frequency domain. Receiver operating characteristic curve analysis of a discriminant function demonstrated significant differences for PLS as compared to time domain indices. An important finding was the significance of MI locus in the time domain indices. PLS did not exhibit this dependence. These data suggest that the usual indices are insufficient for identifying AMI patients at risk of VT. PLS, on the other hand, is valuable regardless of MI location.

[The hygienic status in community health nursing stations]. / Zur hygienischen Situation in Gemeindeschwesternstationen.

199 bases of so-called "Gemeindeschwestern" (nurses delegated by communal authorities as medical auxiliary personnel to practising physicians) in the territory of the former district of Neubrandenburg (now part of the new Land called Mecklenburg-Vorpommern) were examined to determine the sanitary and hygienic conditions obtaining in these bases. Many of these were found to be unsatisfactory: not only the desolate state of the building materials, but also the available space and the water supply. In 14.6% of the consulting rooms hands could not be washed in running water. 39 nurses were found to be violating the rules governing the handling of drugs in respect of storage practices. Sterilisation equipment was often obsolete, and some of the nurses were not complying with their obligation to keep records of the performance of the equipment and to conduct obligatory checks on its proper functioning. Other violations concerned improper storage and unsatisfactory packing of the sterilised items. In 20.1% of the cases disinfectants were incorrectly employed. 45.2% of the ready solutions did not conform with the nurses' statements. A multitude of rules governing the safety of working were ignored due to neglect, quite apart from the low overall standard of hygiene; due to all these unpleasant circumstances, the safety upon which a patient must be able to rely, was no longer assured.

Increased inotropic state during splanchnic artery occlusion shock in the dog.

Myocardial function during various forms of shock and its assessment remain the subjects of continuing controversy. Dogs were instrumented for the measurement of a lead II ECG, systemic arterial pressure, LV pressure, LVdP/dt, and LV anterior wall thickness. Afterload was varied with a snare occluder around the aorta. Contractility was assessed by the end-systolic pressure-wall thickness relationship. After taking control measurements, shock was induced by occluding the celiac and the superior and inferior mesenteric arteries for 2 hr followed by reperfusion. The end-systolic pressure-wall thickness relationship was determined at 1 and 2 hr postocclusion and then at 15, 30, 45, and 60 min postrelease and hourly thereafter. The slope of this relationship was either unchanged or increased (ie greater negative slope) during occlusion. Over the first hour of reperfusion of the splanchnic bed the slope averaged 204 +/- 17% of control, and it reached 221 +/- 41% of control by 3 hr postrelease. Myocardial depression was seen only as an agonal event (39 +/- 6% of control). Sham control dogs were stable over a 4 hr period following sham occlusion. Thus, myocardial contractility was increased during splanchnic artery occlusion shock except as a terminal event. Early depression of circulatory performance was a result of decreased venous return and not cardiac dysfunction.

Myocardial depression during acute pancreatitis: fact or fiction?

Clinical and experimental evidence suggests that myocardial depression occurs during severe pancreatitis, but this evidence is derived from techniques that are not optimal for assessing myocardial contractility (e.g., rate of rise in ventricular pressure [dP/dt]). The slope of the left ventricular (LV) and systolic pressure dimension relationship (Ees), a better indicator of myocardial function, has not been measured in pancreatitis. Ten mongrel dogs underwent surgical instrumentation to monitor systemic arterial and LV pressure, cardiac output, LV dP/dt, and anterior LV wall thickness. End of systole was defined by the peak negative dP/dt. The end-systolic points used to calculate Ees were obtained by aortic and vena caval occlusion. After surgical recovery, pancreatitis was induced via cannulation of the pancreatic duct and injection of autologous bile (1 ml/kg) at 200 mm Hg perfusion pressure. All measurements were taken during a control period and daily after pancreatitis was induced. Pancreatitis was confirmed by a significant increase in serum amylase throughout the study and by autopsy finding of hemorrhagic necrosis. Ees was increased throughout the experimental protocol (1 to 7 days) (p less than 0.05). Myocardial performance as assessed by Ees was significantly increased and myocardial depression did not occur in untreated, conscious dogs with severe pancreatitis. Peak positive LV dP/dt was a poor index of contractility during pancreatitis since it decreased while myocardial contractility was increased. Cardiac depression in pancreatitis noted in other reports was likely due to decreased preload and not to intrinsic cardiac dysfunction.

Glucose dyshomeostasis and cardiovascular failure in endotoxic dogs.

Fasted mongrel dogs were anesthetized with pentobarbital sodium and instrumented for the on-line measurement of blood glucose (BG), a lead II electrocardiogram, and pressures in the left ventricle and pulmonary and systemic artery concomitant with on-line monitoring of BG. Serum insulin was measured by radioimmunoassay, and cardiac output (CO) was determined by thermodilution. Stroke work (SW) and pulmonary and systemic resistances were calculated. After a 30-min control period dogs were treated with Escherichia coli endotoxin (E) or normal saline (S) and then observed for 10 h or until death. Preinjection control BG was maintained in S dogs, and early hyperglycemia (H) was observed in six dogs; in contrast 10 E dogs showed no hyperglycemia (NH). During the late stages all E dogs were markedly hypoglycemic. In both groups of E dogs an early hyperinsulinemia occurred. CO and SW were depressed in both groups of E dogs. These variables were significantly lower in NH than in H dogs. Pulmonary and systemic resistance progressively increased in NH dogs after endotoxin administration. The results suggest that the ability to increase blood glucose levels after endotoxin injection is important for the maintenance of cardiovascular function. Glucose dyshomeostasis leading to hypoglycemia, however, may be a factor in the development of endotoxic cardiovascular failure.

Glucose clamping and cardiovascular function in endotoxic dogs.

Fasted mongrel dogs were anesthetized with pentobarbital sodium and instrumented for the continuous measurement of blood glucose (BG), a lead II electrocardiogram, and pressures in the left ventricle (LV), pulmonary artery, and aorta. Cardiac output was measured every 15 min using thermodilution and LV stroke work, and pulmonary and systemic resistances were calculated. After a 30-min pretreatment period, glucose clamping was initiated. The desired glucose levels were reached within 45 min (hypoglycemic 20 +/- 1 mg/dl, n = 11; normoglycemic 85 +/- 1, n = 7; hyperglycemic 156 +/- 3, n = 7). At this point dogs were treated with either endotoxin (8 mg/kg to 6 hypoglycemic, 4 normoglycemic, and 4 hyperglycemic) or saline (5 hypoglycemic, 3 normoglycemic, and 3 hyperglycemic). All infusions were terminated after 2 h glucose clamping, and all dogs were monitored either until death or for a maximum of 10 h. Hypoglycemic clamping curtailed survival in endotoxic dogs. Hyperglycemic clamping markedly prolonged survival. Normoglycemic clamping left survival time unchanged compared with untreated dogs. The effects of glucose clamping on cardiovascular function during endotoxic shock paralleled the effects on survival. Cardiovascular function was also depressed in hypoglycemic-clamped saline dogs. It is concluded that glucose dyshomeostasis may be a crucial factor in the development of fatal cardiovascular dysfunction and shock after endotoxin administration.

Increased myocardial contractility during endotoxin shock in dogs.

The slope of the left ventricular (LV) end-systolic pressure-diameter relationship (Ees) was analyzed in open-chest, pentobarbital-anesthetized dogs before and after endotoxin administration. A lead II electrocardiogram, systemic arterial pressure, LV pressure, LV dP/dt, and LV minor axis diameter were measured. After control measurements were taken, dogs were given either 1 mg/kg Salmonella enteritidis endotoxin (n = 5) or an equivalent volume of saline (n = 4). Control dogs were followed for 240 min. Endotoxic dogs were monitored until death (246 +/- 44 min). There were no significant changes in Ees in control dogs (17 +/- 3 mmHg/mm), which were hemodynamically stable for 4 h. Ees was significantly increased in endotoxic dogs even into the late stages of shock (41 +/- 11 mmHg/mm, P less than 0.01). Only during the terminal phase did Ees fall significantly below control (11 +/- 2 mmHg/mm, P less than 0.05). End-diastolic diameter decreased following endotoxin administration (P less than 0.05) but returned toward control by the terminal stage. Peak + LV dP/dt was depressed following endotoxin injection. Myocardial contractility was not depressed except as a terminal event. Early depression of cardiovascular performance in endotoxic dogs was therefore due to decreased preload and not cardiac dysfunction.

Hypoglycemic depression of hepatic phagocytosis in vivo and in the in situ perfused rat liver.

Depression of the phagocytic function of the reticuloendothelial system (RES) during endotoxic hypoglycemia has been implicated in the pathogenesis of endotoxin shock. The present study evaluated the in vivo effects of hypoglycemia on RES function and assessed the effects of an vivo bout of hypoglycemia on phagocytosis in the in situ perfused rat liver. Hypoglycemia was produced in male Holtzman rats using either 1 U of regular insulin (RI) (ILETIN, Lilly) or 0.75 U of long-acting insulin (LAI) (85% LENTE/15% ULTRALENTE, Lilly). RES function was quantitated by intravascular clearance of 8 mg/100 gm body weight colloidal carbon (CC). Two hr after RI and 2.5 hr after LAI, the intravascular halftimes of CC clearance were 19 +/- 2 min (N = 22) and 18 +/- 1 min (N = 19), respectively, as compared to control, 11.3 +/- 0.4 min (N = 53, P less than 0.001). The corresponding plasma glucose (PG) levels were 95 +/- 2 mg/dl in control, 14.4 +/- 0.9 for the RI group, and 17 +/- 1 for LAI. Two hr after RI, livers were perfused for 10 min in situ with 50 mg/liter CC in saline 5% rat serum. PG for control liver donors were 90 +/- 3 mg/dl, while those for hypoglycemic liver donors were 15 +/- 2. CC uptake was decreased from 22 micrograms/min/gm liver in the control (+ serum, n = 19) to 11 +/- 2 in hypoglycemia livers (N = 6); no effect of serum on hypoglycemic depression of the RES was seen. There were no differences in flow rates in the 2 groups. These results indicate that hypoglycemia directly impairs RES function and that the in vivo depression of intravascular clearance is not related to either the presence or absence of serum factors or total hepatic blood flow. Thus, the characteristic hypoglycemia of endotoxin shock may contribute to RES depression and the lethal shock syndrome.