The genetic predisposition and its impact on the diabetes mellitus development in patients with alcoholic chronic pancreatitis.

The most common cause of chronic pancreatitis (CP) is alcohol abuse. The aim of the present study was to identify patients with genetic predisposition to CP abusing alcohol. The question posed was whether CP manifests at a younger age and diabetes mellitus develops earlier in individuals with genetic predisposition. The study encompassed 79 patients with alcoholic chronic pancreatitis (ACP) and control group (100 persons). The following mutations were determined: R122H and N29I of PRSS1 and N34S of SPINK1 as well as E366K and E288V of SERPINA 1. No R122H and N291 mutations were observed in the group of ACP patients and in controls. Moreover, there was no E288V mutation. In 79 ACP patients, six SPINK 1 (N34S/wt) mutations were observed. In the control group, one heterozygous SPINK 1N34S gene mutation was found (P = 0.0238). Two PiZ mutations were identified in patients with ACP and one analogical mutation in controls. Amongst patients with ACP as well as SPINK1 and PiZ mutations, the onset of disease was observed earlier and developed earlier. The prevalence of SPINK1 mutation is higher in patients with ACP than in healthy populations. This mutation together with the effects of alcohol accelerates the development of ACP and of diabetes mellitus.

Glucocorticosteroids as markers of death from hypothermia.

In the course of hypothermia, biochemical changes occur that are associated with stimulation of protective thermogenic mechanisms as well as mobilization of internal energy resources mediated by the hormone system. The objective of the investigation was the assessment of validity of determinations of cortisol, cortisone and corticosterone as hypothermia markers in cases of fatal hypothermia combined with concomitant insobriety of the victims. The experimental group consisted of blood samples collected in the course of medico-legal autopsies of 23 hypothermia victims. The controls included blood samples originating from 34 victims of violent sudden deaths (deaths by hanging and traffic road accidents at the scene) and from ten individuals deceased after prolonged agony in consequence of post-traumatic subdural hematomas. In both groups, three subgroups were distinguished that included cases with ethanol levels within the following ranges: 0.0-0.99, 1.0-2.99 and ≥3.0‰. The comparison of determination results showed that irrespectively of blood ethanol concentration, cortisol, cortisone and corticosterone levels seen in hypothermia victims were significantly higher as compared to the controls (P<0.001).

Low dose of bupropion significantly enhances the anticonvulsant activity of felbamate, lamotrigine and topiramate in mice.

Experimental evidence indicates that bupropion hydrochloride, an antidepressant and a first-line smoking cessation aid, exerts dose-dependently anticonvulsant and convulsant effects. In this study, chronic bupropion pretreatment intraperitoneally (i.p.) for 14 days in a dose of 5 mg/kg reduced the ED(50) (i.e. the dose protecting 50% of mice against electroconvulsions) of lamotrigine, topiramate, and felbamate from 4.58, 60.95, and 48.79 (antiepileptic+vehicle) to 3.01, 41.68, and 37.28 mg/kg (antiepileptic+bupropion), respectively, against maximal electroshock-induced seizures in mice. Bupropion significantly increased the plasma and brain concentrations of lamotrigine. Plasma concentration of topiramate was elevated, however, the brain concentration of the drug was not affected. Neither plasma nor brain concentrations of felbamate were elevated by bupropion administration. Bupropion did not exacerbate motor coordination impairment caused by the antiepileptic drugs in the rotarod test. Chronic administration of bupropion significantly potentiates the protective activity of lamotrigine, topiramate, and felbamate against maximal electroshock-induced seizures. A pharmacokinetic interaction is responsible for the effect of bupropion co-administered with lamotrigine.

Pharmacodynamic and pharmacokinetic interactions between common antiepileptic drugs and acetone, the chief anticonvulsant ketone body elevated in the ketogenic diet in mice.

PURPOSE: Acetone is the principal ketone body elevated in the ketogenic diet (KD), with demonstrated robust anticonvulsant properties across a variety of seizure tests and models of epilepsy. Because the majority of patients continue to receive antiepileptic drugs (AEDs) during KD treatment, interactions between acetone and AEDs may have important clinical implications. Therefore, we investigated whether acetone could affect the anticonvulsant activity and pharmacokinetic properties of several AEDs against maximal electroshock (MES)-induced seizures in mice. METHODS: Effects of acetone given in subthreshold doses were tested on the anticonvulsant effects of carbamazepine (CBZ), lamotrigine (LTG), oxcarbazepine (OXC), phenobarbital (PB), phenytoin (PHT), topiramate (TPM) and valproate (VPA) against MES-induced seizures in mice. In addition, acute adverse effects of acetone-AEDs combinations were assessed in the chimney test (motor performance) and passive avoidance task (long-term memory). Pharmacokinetic interactions between acetone and AEDs were also studied in the mouse brain tissue. RESULTS: Acetone (5 or 7.5 mmol/kg, intraperitoneally [i.p.]) enhanced the anticonvulsant activity of CBZ, LTG, PB, and VPA against MES-induced seizures; effects of OXC, PHT, and TPM were not changed. Acetone (7.5 mmol/kg) did not enhance the acute adverse-effect profiles of the studied AEDs. Acetone (5 or 7.5 mmol/kg, i.p.) did not affect total brain concentrations of the studied AEDs. In contrast, VPA, CBZ, LTG, OXC, and TPM significantly decreased the concentration of free acetone in the brain; PB and PHT had no effect. CONCLUSIONS: Acetone enhances the anticonvulsant effects of several AEDs such as VPA, CBZ, LTG, and PB without affecting their pharmacokinetic and side-effect profiles.

Acetonaemia as an initial criterion of evaluation of a probable cause of sudden death.

The archived head-space chromatograms of ethanol determinations in autopsy blood in the years 1996-2003 were analysed. One hundred and two cases with elevated acetone level >250mmol/l were selected in which the biochemical profiles of volatile alcohols (methanol, isopropanol and n-propanol) were determined after "post-hoc" calibration of the constant internal standard. Based on the files obtained from the Prosecutor's Office, the circumstances of death and those preceding death (alcoholism, prolonged or single consumption of alcohol, intoxications with other substances, hypothermia, undernourishment, diabetes) were analysed and the most probable cause of endogenous or exogenous ketonaemia were determined. All cases of unexplained deaths in alcoholics with the ethanol concentration <0.4g/l occurred after withdrawal of long-term consumption of alcohol while all alcoholics with the ethanol concentration >0.4g/l died during the so-called drinking bout. In the group of hypothermia-related deaths with ethanol concentrations <0.4g/l, the acetone concentration was statistically significantly higher than that in hypothermia group with ethanol concentration >0.4g/l in which "congeneric" concentrations of methanol and isopropanol were additionally observed. Furthermore, an algorithm of further diagnostic management was suggested to distinguish the most likely origin of acetonaemia, i.e. accumulation of exogenous "denaturants" of alcohol consumed and cases of endogenous ketogenesis.

[Diagnostic usefulness of the beta-hydroxybutyrate/acetone ratio in medico-legal diagnostics of sudden deaths]. / Ocena przydatnosci diagnostycznej wspólczynnika beta-hydroksymaslan/aceton w sadowo-lekarskiej diagnostyce naglych zgonów.

Our previous studies demonstrated the usefulness of screening determinations of acetone as an initial diagnostic criterion in deaths due to hypothermia, alcoholic ketoacidosis, diabetes mellitus, starvation and some poisonings. In alcoholemia, particularly in cases of prolonged ethanol consumption, the above-mentioned conditions may not result in acetonemia, despite marked concentrations of beta-hydroxybutyrate acid (beta-HBA). Therefore, for the purpose of the present study, the method of beta-HBA determination was modified using GC-MS-EI and applied to analyze 47 autopsy blood samples of individuals who died suddenly due to unknown causes. In 15 cases, the concentration of beta-HBA was higher than 1000 micromol/l; in six subjects from this group, the acetone concentration was lower than 250 micromol/l. In some cases, thus, the use of beta-HBA as an additional diagnostic criterion allows for explaining the pathomechanism of premortal metabolic disturbances.

In vitro co-metabolism of acetoacetate and ethanol in human hepatic mitochondrial and cytosolic fractions.

The rate of alcohol elimination is highly resistant to acceleration in vivo in well-nourished individuals. The acceleration of ethanol elimination may be achieved by providing the conditions in which the action of alcohol dehydrogenase is not delayed by the insufficiency of the oxidized NAD form. The aim of the study was to verify the theoretically assumed mechanism of accelerating alcohol elimination by administering excessive acetoacetate (Ac-Ac) in the experimental in vitro model. Ac-Ac forming the redox system with beta-hydroxybutyrate (beta-HBA) is the natural acceptor of excessive protons from ethanol oxidation. Ac-Ac and beta-HBA penetrate freely through the cell membranes and are easily assimilated energetic substrates. The examinations were performed using the hepatic homogenates (collected from the cadavers shortly after death) supplemented with ethanol and Ac-Ac. The ethanol levels were determined at 0, 15, 60, 90 and 150 min of the experiment. The findings showed that the equimolar addition of Ac-Ac resulted in a two- to three-fold increase in ethanol oxidation in hepatic homogenates. The biochemical system discussed above resembles the natural way of utilizing the excessive NADH, which is formed during ethanol combustion in chronic alcoholics. The results indicate that further investigations are necessary to assess the clinical importance of this metabolic system.

Biochemical background of ethanol-induced cold susceptibility.

The process of cooling is always associated with the depletion of energetic reserves and burning the ketone bodies covers the tissues' needs. Ethanol shows antiketonaemic effects changing the cellular redox potential, inhibiting beta-oxidation of fatty acids, stimulating the release of insulin and inhibiting the release of its antagonist. The aim of the study was to determine whether the cooling process of the organism in the presence of ethanol intoxication may be related to inhibition of the physiological mechanism of ketogenesis induced by hypothermia. The study involved the 67 autopsy cases from 1996 to 2002, in which the circumstances of death indicated the effects of overcooling. This was confirmed on the basis of the data from the Prosecutor's Offices. Then, the chromatograms of autopsy blood alcohol determinations were analyzed and the acetone levels recorded. The analysis supported the hypothesis that the severity of ketosis is inversely proportional to the blood ethanol concentration. Furthermore, it demonstrated that signs of prolonged cold exposure were less frequently observed in unsober persons (frostbites, gastric hemorrhages). Increased sensitivity of intoxicated individuals to cold may be related not only to the dilation of the peripheral vessels, inhibition of shivering thermogenesis caused by muscle relaxation, central nervous system depression and behavioral factors but also to the antiketonaemic effects of ethanol.

[Post mortem diffusion of carbon monoxide to muscles and blood--preliminary examinations]. / Posmiertna dyfuzja tlenku wegla do miesni i krwi--badania wstepne.

The purpose of the study was to determine whether the postmortem diffusion of carbon monoxide (CO) significantly affected the results of carboxyhemoglobin (COHb) and carboxymyoglobin (COMb) determinations. The musculocutaneous and muscular specimens collected from adult cadavers were used. The specimens were treated with CO for 24 h at room temperature. COHb and COMb were determined using gas chromatography. It was demonstrated that the skin substantially limited the diffusion of CO which slightly penetrated only the superficial layers of the muscle and did not change the blood level of COHb in the 4.5-cm layer of the muscle located underneath. The CO diffusion through the superficially charred and thermally coagulated muscle did not differ from that observed in the intact integuments. On the other hand, the membrane of the skin completely deprived of the adipose layer was not the barrier to moderate diffusion into the blood layer situated below. Thus, in charred corpses the results pf COHb and COMb determinations in the material collected under the layer of charred and coagulated tissues enable us to determine whether the victim was alive at the moment of the outbreak.

Stability of toluene and reduction of acetone to 2-propanol in homogenates of the human liver, brain and lungs.

The homogenates of the livers, lungs and brains collected from five different cadavers were placed in the desiccator filled with vapours of rubber glue solvents and the concentrations of toluene, acetone and 2-propanol were determined during the 28-day storage at +25, +4 and -20 degrees C. It was demonstrated that only freezing of the material stabilised the initial concentration of these three xenobiotics while cooling to +4 degrees C resulted in limited conversion of acetone to 2-propanol and additionally reduced the biodegradation of toluene in the brain homogenates. Moreover, it was showed that at +25 degrees C the loss of acetone was almost equimolarly balanced by the 2-propanol increase, which allowed to estimate the initial concentration of acetone with the mean error of about 10%.

Genetic data on 19 STR loci in south-east Poland.

Allele frequencies for the 19 STRs loci, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, CSF1PO, F13A01, F13B, FESFPS, FGA, LPL, Penta D, Penta E, TH01, TPOX and VWA were obtained from a sample of 203-1188 unrelated individuals living in the area of south-east Poland.

Evidential value of injuries useful for reconstruction of the pedestrian-vehicle location at the moment of collision.

The paper presents final results of the studies concerning the usefulness of knee, ankle, hip, pelvis and neck injuries in reconstructing the circumstances of car-to-pedestrian accidents. Each type of injuries was evaluated with regard to possible reconstruction of the victim's position (upright or recumbent) at the moment of collision and in the upright hits-the side of the pedestrian's body hit. In each group, a chance of proper reconstruction of the pedestrian's location (which determined the frequency of injury pattern assumed as the typical one of a given position or impact side) and error risk (percentage of cases in which the injury pattern showed improper position or impact side) were calculated. These data were compared with similarly calculated possibilities of deducing based on the classical "bumper" injuries to soft tissues and "bending" fractures of the lower limb bone diaphyses. It was shown that the bone bruises within the knee epiphyses were very specific evidence for upright hits as they confirm the limb load by body mass at the moment of pathological dislocation of joint structures on impact. The evidential value of knee injuries was found to be similar to that of other "classical" methods based on bumper injuries or even higher in lateral and front hits. The injuries to the remaining structures were less frequently found and their correlation with the victim's position or impact side was lower. Nevertheless, once the whole complex of these injuries is taken into consideration the chances of proper reconstruction of the pedestrian-vehicle location increase and the risk of opinion error is minimized.

The influence of ethanol on the level of ketone bodies in hypothermia.

Presently available possibilities of macro- and microscopic diagnosis of death from hypothermia are very limited as the changes observed are either weakly specific (ecchymoses in the mucous membrane of the stomach, histological features of haemorrhagic pancreatic necrosis, cardiomyocyte necrosis or decreased content of glycogen in hepatocytes) or represent only local action of low temperatures (frostbites, violet patches in the region of knees and elbows, red livores) and they may not be present in cases of death from cooling at environmental temperature close to zero or higher. The study evaluated the usefulness of acetoacetic acid (Ac-Ac), beta-hydroxybutyric acid (beta-HBA) and acetone determinations in blood, urine and vitreous humour for diagnosis of death from hypothermia. These three substances called ketone bodies, are easily assimilated energetic substrates that get oxidized preferentially before glucose and fatty acids. In hypoglycaemia (also hypothermia-induced one), the tissues dependent on glucose (e.g. the brain) cover most of their energetic needs from oxidation of these compounds. The analysis of 16 cases of death in circumstances suggesting hypothermia (mainly of the alcohol abusers) showed that the degree of ketosis was inversely proportional to the blood ethanol concentration. This relation may result from stimulation of insulin release and a decrease in the release of its antagonists by ethanol, as well as from inhibition of free fatty acid (FFA) beta-oxidation due to increase in the NADH/NAD ratio. So, the antiketonaemic effects of ethanol (together with its influence on the dilatation of the peripheral vessels and inhibition of shivering thermogenesis by muscle relaxation), explain increased sensitivity of intoxicated persons to low temperatures.