Prediction of adrenocortical insufficiency after pituitary adenoma surgery using postoperative basal cortisol levels.

Our aim was to analyze the correlation of early postoperative cortisol levels in patients after transsphenoidal pituitary adenoma surgery compared to the standard dose ACTH test and Insulin tolerance test (ITT) several months later. We retrospectively reviewed data from 94 patients operated for pituitary adenoma in years 2009-2012. The comparison of day 7 (median) postoperative basal cortisol levels and 3.6 months (median) after pituitary adenoma surgery stimulation test - standard dose 250 microg 1-24ACTH test in 83 patients or ITT in 11 patients were performed. All 16 patients with early postoperative cortisol levels >500 nmol/l proved a sufficient response in the stimulation tests. At basal cortisol levels of 370-500 nmol/l the sufficient response was found in 96 % (27/28) of patients. In the postoperative basal cortisol levels 200-370 nmol/l we found a preserved corticotroph axis later on in 88 % (28/32) of cases. Patients with basal cortisol levels 100-200 nmol/l had a maintained corticotroph axis function in 8/11 cases - 73 %. All patients with an early postoperative basal cortisol level above 500 nmol/l proved in the stimulation tests a preserved corticotroph axis function. The interval 370-500 nmol/l showed a minimal risk of postoperative adrenal insufficiency.

Impaired microvascular reactivity and endothelial function in patients with Cushing's syndrome: influence of arterial hypertension.

The aim of the study was to evaluate skin microvascular reactivity (MVR) and possible influencing factors (fibrinolysis, oxidative stress, and endothelial function) in patients with Cushing's syndrome. Twenty-nine patients with active Cushing's syndrome (ten of them also examined after a successful operation) and 16 control subjects were studied. Skin MVR was measured by laser Doppler flowmetry during post-occlusive (PORH) and thermal hyperemia (TH). Malondialdehyde and Cu,Zn-superoxide dismutase were used as markers of oxidative stress. Fibrinolysis was estimated by tissue plasminogen activator (tPA) and its inhibitor (PAI-1). N-acetyl-beta-glucosaminidase, E-selectin, P-selectin, and ICAM-1 were used as markers of endothelial function. Oxidative stress and endothelial dysfunction was present in patients with hypercortisolism, however, increased concentration of ICAM-1 was also found in patients after the operation as compared to controls (290.8+/-74.2 vs. 210.9+/-56.3 ng.ml(-1), p<0.05). Maximal perfusion was significantly lower in patients with arterial hypertension during PORH and TH (36.3+/-13.0 vs. 63.3+/-32.4 PU, p<0.01, and 90.4+/-36.6 vs. 159.2+/-95.3 PU, p<0.05, respectively) and similarly the velocity of perfusion increase during PORH and TH was lower (3.2+/-1.5 vs. 5.2+/-3.4 PU.s(-1), p<0.05, and 0.95+/-0.6 vs. 1.8+/-1.1 PU.s(-1), p<0.05, respectively). The most pronounced impairment of microvascular reactivity was present in patients with combination of arterial hypertension and diabetes mellitus.

[Correlation among clinical and morphological findings in a diffuse axonal injury]. / Korelace klinických a morfologických nálezu u difuzního axonálního poranení.

In a man of 56 years--a victim of traffic accident, contusions in both temporal lobes of the brain and a small subdural and subarachnoid haemorrhage were diagnosed by CT and MR examination. According to the findings of petechial haemorrhages in the corpus callosum a diffuse axonal injury was clinically diagnosed. After 2 months the patient died. Excisions of corpus callosum and the brain-stem were examined by histological staining including Palmgren@s method and by immunohistochemical detection of neuron-specific enolase (NSE) in damaged axons. Clinical findings were compared with morphological findings, i.e. autopsy, histological and immunohistochemical examination. The possibilities of clinical diagnostics of diffuse axonal injury are discussed.

[Trauma of cervical spinal cord--an immunohistochemical study]. / Poranení krcní míchy--imunohistochemická studie.

The histologica and immunohistochemical examination of cervical spinal cord were used in the group of 11 deceased of violent death. The positivity of neuron-specific enolase (NSE) in axons of cervical spinal cord was examined, and findings in the group of deceased on craniocervical injury (7 cases) and the group of deceased without cranio-cervical trauma (4 cases) were compared. While a routine histological examination including Palmgren's staining showed no significant differences between bouth groups, we noticed an increased frequency of appearance of NSE deceased with craniocervical injury.

[Effect of hypercortisolism on development of atherosclerotic changes in blood vessels]. / Vliv hyperkortizolismu na vznik a vývoj aterosklerotických cévních zmen.

UNLABELLED: Hypercortisolism is associated with a high risk of sickness rate and death rate particularly in view of facilitated arteriosclerotic processes. It is most frequently induced by drug therapy, but endogenous hypercortisolism (Cushing's syndrome) may serve as a suitable model of the effect of hypercortisolism on vascular wall. Our cohort included the following groups of patients and control individuals: 1. a group of patients with florid so far untreated Cushing's syndrome--14 patients, 2. a control group to these patients--16 individuals, 3. a group recently operated on and healed-up patients with Cushing's syndrome--8 patients, 4. a group of previous of previous cured-up patients with Cushing's syndrome--27 subjects, 5. a control group to those patients of group 4--17 persons. The following differences were found between the respective groups: 1. the ultrasonographic examination of carotid arteries demonstrated sclerotic plates or carotid stenosis in 21.3% of patients with florid Cushing's syndrome and 41.4% in patients with corticolism having been cured-up against 11.7% in the relevant control group; 2. the examination of skin microcirculation by the laser-doppler method revealed a lower velocity of perfusion increase during examination of postocclusion hyperemia in patients with florid Cushing's syndrome and hypercortisolism having been cured-up against a control group (CUSH., P < 0.04; previous cured-up, P < 0.02) as well as thermally-induced hyperemia (CUSH., P < 0.03; formerly cured-up, P < 0.04); 3. the laboratory examination of patients with florid Cushing's syndrome revealed higher values of LDL-cholesterol (P < 0.05) and total cholesterol (P < 0.001), malonyldialdehyde as an indicator of increased formation of oxygen radicals (P < 0.05) and oromucoid, the protein of acute phase, signaling a chronic inflammation (P < 0.05); 4. in patients who previously suffered from hypercortisolism increased levels of fibrinogen (P < 0.03) and the cytoadhesive molecule ICAM-1 (P < 0.05) were accompanied by decreased levels of the growth factor of vascular endothelia (VEGF) (P < 0.05) against patients with florid Cushing's syndrome. CONCLUSION: The findings of the examinations performed indicate that increased incidence of arteriosclerotic processes is present in patients with the florid Cushing's syndrome as well as in those who have suffered from Cushing's syndrome before.

[Endothelial, phospholipid and oxidized LDL antibodies in patients with endogenous hypercortisolism]. / Protilátky proti endoteliím, fosfolipidum a oxidovaným LDL u pacientu s endogenním hyperkortizolismem.

Antiphospholipid (APLA), antiendothelial (AECA) and anti-oxidized LDL (anti-oxLDL) autoantibodies are found in vascular disorders. Pathogenetic contingency of atherosclerosis and these autoantibodies is still discussed, the mechanisms of their action in atherogenesis are not quite clear so far. Patients in various stages of endogenous hypercorticism as a model of accelerated atherosclerosis were investigated. We have sought possible correlations between autoantibodies and parameters of atherosclerosis with regard to the influence of endogenous hypercorticism on the inflammation. Low titres of autoantibodies in patients with active forms of disease result from the immunosuppressive effect of steroids. None of investigated group had high titres of APLA. No differences were found in AECA occurrence. No correlation of APLA, anti-oxLDL nor AECA with urinary free cortisol and plasma cortisol was found. There were no significant differences in autoantibody titres between patients with or without carotid stenosis. These results suggest, that autoantibodies may not always influence the development and progression of atherosclerotic lesions.

Immunostimulatory effect of Bacillus firmus on mouse lymphocytes.

Bacillus firmus (a Gram-positive nonpathogenic and harmless bacterium), was shown to be a strong polyclonal activator of mouse B lymphocytes as estimated by ELISA testing of Ig concentrations in culture supernatants after incubation of BALB/c mouse splenocytes with inactivated bacillus. Synthesis of all main Ig classes and all IgG subclasses was stimulated in vitro, the considerable effect on IgA formation being the most interesting feature. B cell stimulation was T cell dependent, as was demonstrated by the effect of B. firmus on all Ig isotypes and by comparison of lymphocyte response of nu/nu mice and heterozygous nu/+ mice. The effect of B. firmus on splenocyte proliferation was stimulatory or suppressive depending on the dose of the bacterium. Increased synthesis of IFN-gamma and IL-10 (detected by ELISA in splenocyte culture supernatants) showed probable stimulation of Th1 and Th2 subpopulations. Considering the stimulatory effect on IgA formation and macrophage stimulation, B. firmus seems to be a prospective mucosal adjuvant and/or probiotic.

[Detection of metalloproteinases in pulmonary tissue after sudden and violent death]. / Prukaz metaloproteináz v plicní tkáni u náhlé a násilné smrti.

During early stages of chronic hypoxia under experimental conditions in the lungs alveolar macrophages are activated and in the latter an increased amount of matrix-metalloproteinase-collagenolytic enzymes is produced. Their presence was assessed in the present study in the lungs of infants who died suddenly (SIDS) and also in subjects who died from a violent death incl. acute and protracted external suffocation. The positive findings of these matrixins, in particular MMP-9 (gelatinase) were assessed in alveolar macrophages not only after protracted suffocation but also in the majority of sudden infant deaths. These findings supplement the mosaic of changes which indicate that sudden infant deaths are preceded by a period when the infant was exposed to hypoxia.

[Effect of autolysis on histochemical examinations of the central nervous system]. / Vliv autolýzy na imunohistochemické vysetrení CNS.

The objective of the study was to explain the effect of autolysis on immunohistochemical detection of neurone-specific enolase (NSE), beta-amyloid protein precursor (beta-APP) and ubiquitine in cerebral tissue. The examination was made in 6 deceased subjects without mechanical injury of the CNS and 6 subjects with a craniocerebral injury who survived from 6 hours to 3 days. In all deceased subjects the post-mortem examination was made within 24 hours after death. For immunohistochemical examination tissue excisions were taken from standard sites of the brain. The first tissue excisions were immersed into 10% formol after a post-mortem interval of 24 hours. The remaining tissue slices were subjected to autolysis at room temperature and gradually immersed into formol after 24-hour intervals, the longest post-mortem interval being 168 hours, i.e. 7 days. For visualization of the linked primary antibody the biotin-streptavidin system labelled with alkaline phosphatase was selected. In the group of 6 subjects who died after a craniocerebral injury in 4 instances axonal lesions were detected, i.e. axonal oedema or formation of retraction spheroids. The damaged axons were positive on examination with all investigated antibodies, whereby it was possible even after a 168-hour post-mortem interval to differentiate damaged and not damaged axons. In the group of 6 subjects without mechanical injury of the CNS in 5 instances axonal oedema was found, however, it was not positive with anti-NSE antibodies nor with anti-beta-APP. After the 24-hour post-mortem interval in this group in 3 instances ubiquitine positivity was found in axons but already after a post-mortem interval exceeding 2 days the axons were ubiquitine positive in all 6 subjects. Lumpy deposits of this substance could be detected in axons also beyond axonal structures.

[Detection of caspase as a sign of tissue damage]. / Detekce kaspázy jako príznaku poskozené tkáne.

Caspases are cysteine proteases which participate in different stages of apoptosis. Apoptosis, cell death, programmed by its nucleus is associated also with a number of diseases and tissue damage. To this process increasing attention is paid also in the sphere of forensic medicine, in particular to make use of the diagnostic contribution in investigation of cause of death, vital reaction and time when the injury developed. In damaged tissues by immunohistochemical methods caspase activity was assessed. Caspase 8 activity (Flice) was detected in the heart muscle in congenital heart disease in a two-week old infant but also in ischaemia in a 58-year old woman with thrombosis of the coronary artery or in acute circulatory failure after an overdosage of pervitin in a 39-year-old drug addict. In the liver there was a positive finding in hepatocytes in chronic inflammatory changes caused by chronic alcohol abuse. An early caspase 8 reaction after an injury is suggested by positive findings in skeletal muscles of the neck and larynx in a 47-year-old man who was strangled. A positive reaction was proved only at the site of the line caused by the strangulation tool. However macroscopically nor by common histological staining the muscle tissue did not display any signs of injury at the mentioned sites. The results suggest that immunohistochemical evidence of caspase 8 can be used as a suitable complementary examination not only for detection of damaged tissue but also for evaluation of early stages of the vital reaction.

[Glycophorin in forensic medicine]. / Vyuzití glycophorinu v soudním lékarství.

Immunohistological evidence of glycophorin from paraffin sections of tissues fixed with formol, makes its possible to visualize membranes of preserved erythrocytes and fragments after their disintegration. This fact permits even after development of post-mortem changes to differentiate actual extravasation, manifested by a positive finding of glycophorin from mere infiltration of tissue by blood pigment after death, where glycophorin is lacking. The method is very sensitive and can reveal even very discrete haemorrhage. From different grades of erythrocyte disintegration in haematomas in the same subject sometimes conclusions can be drawn also on the different time of development of traumatic tissue changes.

Studies on the potentiometric thallium(III)-selective carbon paste electrode and its possible applications.

Construction, performance characteristics and applications of a carbon paste thallium(III) ion-selective electrode are described. The electrode, which is based on ion-associate compounds formed between cetylpyridinium and chlorothallate(III) complexes dissolved in tricresyl phosphate as pasting liquid, showed near-Nernstian response over the concentration range of 5.8 x 10(-6) - 2.9 x 10(-3) mol/L. Potentiometric titrations of thallium(III) with cetylpyridinium chloride were affected by higher concentrations of excess halides, probably due to the formation of higher halogenothallates.

[Determination of the ABH blood group system in the placenta]. / Príspevek k urcování krevnich skupinových vlastností systému A B H v placente

Blood group substances A, B, H were detect in placental tissue paraffin cuts (after 10% formaline fixation) by immunohistochemistry using indirect immunoperoxidase-two-layers technology or biotin-streptavidin complex signed by alkaline phosphates. Both fetal and maternal part of placenta are to be investigated which enables their group markers to be identified. Immunohistochemical detection of the A, B, H blood group substances in placenta can give conclusion about the blood group of fetus and of fetus and of mother as well.

Some aspects of the use of heteropoly anions in elemental analysis by simple potentiometric ion-pair formation-based titration.

The possibility of the use of heteropoly anions formed by addition of phosphate to the solutions of either molybdate or tungstate in the determination of phosphorus by the simple method of potentiometric titration was studied. The heteropoly anions are titrated on the ion-pairing principle using a titrant containing a lipophilic counter-ion, i.e. 1-(ethoxycarbonyl)pentadecyltrimethylammonium bromide (Septonex), the titration being monitored by carbon paste electrode, although other liquid membrane-based electrodes can also be used. Calibration plots of the titrant end-point consumption vs. concentration are not linear, but allow one to evaluate the content of phosphorus. Similar procedures, when optimized, should be elaborated for determination of numerous other elements forming heteropoly anions.

[Detection of free histamine and serotonin in the evaluation of the vital reaction in injuries]. / Prukaz volného histaminu a sérotoninu pri posuzování vitální reakce poranení.

Skin injury alive produces quick and distinct increase of free histamine and serotonin. A fluorimetric method for detection of these compounds in tissues has been the only one described in the literature up to now but it needed a time consuming extraction and big amount of primary material (about 3 g of skin devoid of subcutaneous fat tissue) as well. The authors succeeded in substituting the method and developed a voltametric detection of histamine and serotonin using the computerized ECO-TRIBO-Polarograph. It enabled to omit the extraction of skin compounds and to decreased the amount of primary tissue to a quarter. The detection of free histamine and serotonin levels is presumed to enable a more accurate determination of the time of injury, particulary in short interval before death; it presents, with a simultaneous of glycophorin, a convenient complementation of immunohistochemical investigation of fibronectin.

[Evaluation of the vital reaction in injuries]. / Posuzování vitality vzniku poranení.

Evidence of an early vital reaction after a skin injury is sometimes of decisive importance in forensic practice. Attention is devoted increasingly to immunohistochemical evidence of substances, the presence of which in the borders of the wounds signalizes an intravital origin of the injury. In this context evidence was focused on detection of glycophorine at sites of haematomas of the skin and subcutaneous layer and to fibronection in the borders of injuries and in haematomas. To visualize the two substances in paraffin tissue sections, the immunohistochemical method of indirect immunoperoxidase reaction was selected where in the first layer specific sera were used wither against glycophorine A, B or against fibronectin. Glycophorine is a sialoglycoprotein which is part of the red cell membrane. It can be detected at the site of haematomas and after the disintegration of erythrocytes during post-mortem autolysis. Conversely it is lacking where membrane structures were not present. This makes it possible even during advanced stages of post-mortem changes to differentiate actual haematomas from mere post-mortem inhibition of tissue by blood pigment (e.g. in hypostasis) where glycophorine cannot be detected. Fibronectin, another glycoprotein substance can, already in the early stage of injury, form the basis for a netlike structure in the shape of fine "strings". Detection of fibronectin suggests the intravital development of injury and in the authors cases it could be detected already within several minutes after the development of the injury.

[Immunohistochemical detection of ABH blood group factors in hair]. / Imunohistochemický prukaz krevních skupinových substancí A B H ve vlasu.

Commonly used serological methods i. e. absorption-elution test and mixed agglutination do not always allow discriminate the nature of blood group substances A, B, H in hair investigation. The finding need not sometimes be unequivocal. A completion of forensic traditional serological methodology by indirect immunoperoxidase technique enables to eliminate doubts about interpretation of results. There is possible to identify blood group substances A, B, H in preserved medullary core of hair in this way.

[2-dimensional echocardiography in the evaluation of right ventricular volume and function. Comparison with echocardiography]. / Dvourozmerná echokardiografie pri hodnocení objemu a funkce pravé komory. Srovnání s echokardiografií.

Using two-dimensional echocardiography (ECHO), the authors examined within one week preceding right-sided catheterization of the right ventricle (RV) 44 patients. Of three tested ECHO techniques for calculation of right ventricular volumes and/or function the best correlations were obtained by the method of evaluation of area-length of the right ventricle from two orthogonal projections (apical 4-cavity and subcostal with visualization of the outflow portion of the RV). The following correlations were assessed: telediastolic volume--r = 0.831, telesystolic volume--r = 0.815 stroke volume--r = 0.810 and ejection fraction (EF)--r = 0.752 (p less than 0.001 for all correlations). The tested method enables us at the same time to screen at least existing right ventricular dysfunction (sensitivity 0.68; specificity 0.82). In the presence of tricuspidal insufficiency (TI) increase in right ventricular function occurs which may mask its existing failure (dependence of EF on the mean pressure level in pulmonary artery was proved only in patients without TI--r = 0.594). We assume that information on right ventricular function is important not only in diseases affecting primarily the right heart but also in advanced stages of left-sided failure. Follow-up of the natural course of the disease and the effect of provided treatment by the non-invasive ECHO method is thus useful not only in research but also in clinical practice.

Three echocardiographic methods in right ventricular function evaluation.

We employed two-dimensional echocardiography for the assessment of right ventricular (RV) volumes and/or function in a series of 44 patients. The results of three different echocardiographic approaches were compared with the data obtained from single-plane RV angiography following ultrasound within a 7-day interval. Only the echocardiographic area length method with two orthogonal imaging planes employed (apical 4-chamber and subcostal projections) yielded the beneficial results. The correlations between echocardiographic and angiographic RV volume estimates were rather high (end-diastolic volume: r = 0.83, end-systolic volume: r = 0.82, stroke volume: r = 0.81) and satisfactory in ejection fraction (r = 0.75). Using the method mentioned, the differentiation of patients with an angiographic evidence of RV failure (echocardiographic ejection fraction less than 0.55) from those without it was possible with a sensitivity of 0.68 and a specificity of 0.82. Concerning the clinical impact of the presented study, we can recommend the technique in question as a screening procedure for the detection of changes in RV function exceeding 12% (95% confidence limits).

The effect of acute and chronic nitrate administration in precapillary and postcapillary pulmonary hypertension.

The effect of nitrates was monitored in 16 patients with precapillary, and in 12 patients with postcapillary pulmonary hypertension (PH). The patients had haemodynamic examination on acute administration of 10 mg of isosorbide nitrate (ID) in infusion and after 2-month therapy with 120 mg of isosorbide dinitrate retard daily. Acute ID administration decreased filling pressure of both ventricles, pulmonary and aortic pressure, pulmonary and systemic resistance and increased blood flow. No significant difference was found between the two types of PH. Long-term administration resulted in a statistically significant decrease in filling pressure (in both ventricles) and pulmonary pressure, even though the decrease was less pronounced than those seen following acute administration. Again, no statistical differences were noted between the two types of PH. While acute administration of nitrates decreases both preload and afterload, chronic administration decreases only preload. The implications of this finding for therapeutic practice are discussed.