PCR-based identification of drowning: four case reports.

Proper diagnosis in drowning victims is often difficult due to the lack of signs specific to drowning. The diatom test is a widely used procedure for the diagnosis. Some types of water contain only minimal amounts of diatom cells which may provide false-negative results, while a negative diatom test result does not exclude drowning. In proving drowning, we used a polymerase chain reaction (PCR)-based biological method in addition to the conventional methods. DNA was extracted from postmortem spleen tissues and water of the drowning site. Samples were tested with algae (diatoms and small green algae)- and cyanobacteria (blue-green algae)-specific primers. We present here multiple drowning cases in which diatom tests of the postmortem tissue samples and the water were negative. In each case, the presence of phytoplanktonic DNA strengthened the autopsy diagnosis of drowning even in the absence of visible diatoms. In the future, the PCR method may be of consideration as a possible supplement of the diatom test in the examination of presumed drowning cases.

Drowning-related fatalities during a 5-year period (2008-2012) in South-West Hungary--a retrospective study.

The purpose of the present study was to investigate all the drowning-related cases in South-West Hungary between 2008 and 2012. It is a retrospective and descriptive study of 114 drowning-related deaths during that 5-year period. The investigation includes both unintentional and intentional drowning cases. We found that the most considerable risk factor of drowning in this area of Hungary is alcohol consumption, because in more than half of the cases the victims were under the influence of alcohol. We also concluded that more than two third of the cases included males. The cause of drowning in younger victims was mostly accident. In both genders subjects aged 50-70 had the highest risk of drowning in the period investigated. Seniors frequently committed suicide by drowning. Drowning deaths occurred in all types of water, mostly in lakes, rivers, canals and other types of catchment in the vicinity of victims' homes (most commonly in wells). This study might help to understand the circumstances and the causes leading to drowning and it may draw the attention to the possible preventive interventions.

[The tumor cured, but what about the patient?]. / A tumor meggyógyult, és a beteg?

BACKGROUND: Polyneuropathy is defined as a simultaneous malfunction of several peripheral nerves, which could be a toxic side effect of cancer therapy; however, this is reported to occur rarely and difficult to confirm. AIMS: The purpose of the study was to introduce, at the first time, the calorimetry in the diagnosis of neuropathy in an experimental animal model. The study was inspired by the forensic investigation of a 53-year-old cancer survivor female patient, in whom the development of polyneuropathy could be caused by cyclophosphamide therapy. METHODS: Adult guinea pigs were injected intraperitoneally with the dose of cyclophosphamide, comparable to the human dosage. Animals were euthanized, then nerve and muscle samples were analyzed using a SETARAM Micro calorimeter. Denaturation temperatures were measured and the calorimetric enthalpies were calculated based on the areas under thermal absorption curves. RESULTS: Thermal denaturations of the samples were decreased and the calorimetric enthalpies were increased depending on the doses of cyclophosphamide. The nerves were more sensitive to chemotherapy as compared to the muscles. CONCLUSION: It is concluded that toxic effects of cyclophosphamide on peripheral nerves and muscles can be measured and analysed by calorimetry, and that the effect was dose dependent.

Differential scanning calorimetry study of glycerinated rabbit psoas muscle fibres in intermediate state of ATP hydrolysis.

BACKGROUND: Thermal denaturation experiments were extended to study the thermal behaviour of the main motor proteins (actin and myosin) in their native environment in striated muscle fibres. The interaction of actin with myosin in the highly organized muscle structure is affected by internal forces; therefore their altered conformation and interaction may differ from those obtained in solution. The energetics of long functioning intermediate states of ATP hydrolysis cycle was studied in muscle fibres by differential scanning calorimetry (DSC). RESULTS: SETARAM Micro DSC-II was used to monitor the thermal denaturation of the fibre system in rigor and in the presence of nucleotide and nucleotide analogues. The AM.ADP.Pi state of the ATP hydrolysis cycle has a very short lifetime therefore, we mimicked the different intermediate states with AMP.PNP and/or inorganic phosphate analogues Vi and AlF4 or BeFx. Studying glycerol-extracted muscle fibres from the rabbit psoas muscle by DSC, three characteristic thermal transitions were detected in rigor. The thermal transitions can be assigned to myosin heads, myosin rods and actin with transition temperatures (Tm) of 52.9 +/- 0.7 degrees C, 57.9 +/- 0.7 degrees C, 63.7 +/- 1.0 degrees C. In different intermediate states of the ATP hydrolysis mimicked by nucleotide analogues a fourth thermal transition was also detected which is very likely connected with nucleotide binding domain of myosin and/or actin filaments. This transition temperature Tm4 depended on the mimicked intermediate states, and varied in the range of 66-77 degrees C. CONCLUSION: According to DSC measurements, strongly and weakly binding states of myosin to actin were significantly different. In the presence of ADP only a moderate change of the DSC pattern was detected in comparison with rigor, whereas in ADP.Pi state trapped by Vi, AlF4 or BeFx a remarkable stabilization was detected on the myosin head and actin filament which is reflected in a 3.0-10.0 degrees C shift in Tm to higher temperature. A similar effect was observed in the case of the nonhydrolyzable AMP.PNP analogue. Differential DSC measurements suggest that stabilization actin structure in the intermediate states of ATP hydrolysis may play an additional role in actin-myosin interaction.