Histopatological parameters of the spinal cord in different phases of experimental autoimmune encephalomyelitis. A mouse model of multiple sclerosis examined by classical stainings combined with immunohistochemistry.

Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE) are characterized by three main histopathological parameters: inflammation, demyelination and axonal damage. In this study, these parameters were assessed in spinal cords of mice in the successive phases of EAE by quantitative histology and immunohistochemistry. The number of inflammatory lesions, the intensity of inflammation and expression of CD45 corresponded with the severity of clinical symptoms: they increased from the onset phase to the peak phase of the disease and subsided in the chronic phase. Demyelination increased in the peak phase and did not change in the chronic phase of EAE, although axonal damage gradually increased from the onset phase to the chronic phase, suggesting compensatory hypermyelination in that phase. The markers of myelin and axonal injury: myelin basic protein (MBP) and beta amyloid precursor protein (ß-APP) showed changes (decrease and increase, respectively) of expression parallel to changes in demyelination and axonal damage. Results of this study indicate that although inflammation intensity subsides in the chronic phase of EAE, the neurodestructive processes: demyelination and axonal damage continue in that phase.

Biomechanical changes in the liver tissue induced by a mouse model of multiple sclerosis (EAE) and the effect of anti-VLA-4 mAb treatment.

Experimental autoimmune encephalomyelitis (EAE) is a mouse model of demyelinating diseases, such as multiple sclerosis (MS). MS can be accompanied by autoimmune hepatitis. In this study, nanomechanical, biorheological and histological examinations were carried out by atomic force microscopy (AFM), rheology, and immunofluorescence microscopy to investigate changes in the liver tissue of EAE mice and the effect of natalizumab, a monoclonal antibody against α4-integrin (VLA-4) cell adhesion molecule, used in MS therapy. Liver samples collected from EAE mice in three successive phases of the disease showed inflammatory changes manifested by leukocyte infiltrations and elevated levels of proinflammatory cytokine IL-1ß. Liver stiffness and viscoelasticity increased in the onset phase of EAE, decreased in the peak phase and increased again in the chronic phase to reach the highest values. These changes were not associated with inflammation parameters which increased in the peak phase and decreased to the lowest values in the chronic phase. Moreover, anti-VLA treatment, which reduced the inflammation parameters, had an ambiguous effect on stiffness and viscoelasticity: it increased them in the peak phase but decreased in the chronic phase. The observed discrepancies can result from a complex network of interactions between inflammation and fibrosis, as well as between liver cells and the extracellular matrix influencing the biomechanical properties of the liver tissue.

Non-ischemic heart preconditioning.

Ischemic heart conditioning has been shown to protect the organ against ischemia/reperfusion injury. Animal studies have revealed that the heart can also be conditioned by non-ischemic procedures, namely physical exercise and tachycardia. Long and short term endurance training, sprint training, resistance or interval training and even one bout of exercise induce cardiac preconditioning, which is manifested by a reduction in post ischemia/reperfusion infarct size, ventricular arrhythmia and improved heart function. Several factors contribute to the exercise-induced heart preconditioning, among which the most important can be: increased activity of the anti-radical defense system, opioids, interleukin-6, nitric oxide, ATP dependent potassium channels, heat shock protein 72 and sphingosine-1-phosphate. A few studies have also shown that one bout of exercise in patients with stable angina increases tolerated workload. According to some data obtained in swine and dogs, stimulated tachycardia before ischemia/reperfusion reduces the infarct size. Future studies are needed to fully clarify the mechanisms responsible for exercise- or tachycardia-induced heart preconditioning against ischemia/reperfusion. It may lead to the development of new treatment modes of the disease.

Effect of tachycardia on mRNA andf protein expression of the principal components of the lipolytic system in the rat's heart ventricles.

There is a convincing piece of evidence showing that most of free fatty acids (FFA) entering cardiomyocytes are first esterified into triacylglycerols (TG) before being oxidized or used for synthesis of complex lipids. The enzyme adipose triglyceride lipase (ATGL) initiates lipolysis of TG by hydrolyzing the first ester bond of the compound. As a result, free fatty acid and diacylglycerol (DG) are released in that process. Finally, DG may be further hydrolyzed by the enzyme hormone sensitive lipase (HSL). The aim of the present study was to examine effect of tachycardia on mRNA and protein expression of ATGL, CGI-58 (an activator of ATGL), G0S2 (an inhibitor of ATGL) and HSL in the left and right ventricle of the rat. The experiments were carried out on male Wistar rats, 240 - 260 grams of body weight. After anesthesia, two electrodes were inserted in the right jugular vein and connected to SC-04 stimulator. The rats were randomly allocated into one of the three groups, namely: control, 30 min and 60 min of the heart stimulation at the rate of 600 times/min. The expressions of ATGL, CGI-58, G0S2 and HSL were evaluated at both gene and protein levels using real-time PCR and Western Blot analysis, respectively. Both 30 and 60 min stimulation reduced ATGL, HSL, CGI-58 and G0S2 mRNA content in the left ventricle. The stimulation had only insignificant impact on ATGL, HSL and CGI-58 transcript levels in the right ventricle. Interestingly, Western Blot analysis showed that the stimulation, regardless of the time, reduced the ATGL and G0S2 protein expression, but did not change the CGI-58 and HSL expression in the left ventricle. Furthermore, in the right ventricle, the protein expressions of ATGL, HSL and G0S2 decreased after stimulation. In conclusion, the obtained results clearly show that tachycardia affects both mRNA and protein expression of particular compounds of the TG lipolytic system in the heart ventricles. Additionally, there are marked differences in the expressions of the examined compounds between the ventricles.

Effect of tachycardia on lipid metabolism and expression of fatty acid transporters in heart ventricles of the rat.

Tachycardia increases oxidation of the plasma-borne long chain fatty acids in the heart. The aim of the present study was to examine effect of tachycardia on: 1) the total level of free fatty acids, diacylglycerols, triacylglycerols and phospholipids in both heart ventricles; 2) (14)C-palmitate incorporation in the lipid fractions; 3) expression of fatty acid and glucose transporters in the ventricles. Tachycardia was induced in anesthetized rats by electrical atrial pacing at the rate of 600/min. Samples of the left (LV) and right (RV) ventricle were taken after 30 and 60 min pacing. The level free fatty acids, diacylglycerols, triacylglycerols and phospholipids was determined by means of gas-liquid chromatography and (14)C-palmitate incorporation by liquid scintillation counting, respectively. Expression of fatty acid- and glucose-transporters was determined using Western blot technique. In LV, 30min pacing increased the content of diacylglycerols whereas the content of other lipids remained stable. After 60 min of pacing the levels of the examined lipid fractions did not differ from the respective control values. In RV, the content of diacylglycerols and triacylglycerols was reduced both after 30 and 60 min pacing. Tachycardia also affected incorporation of (14)C-palmitate in lipid fractions of goth ventricles. 30 min pacing up-regulated plasmalemmal expression of FAT/CD36 (fatty acid translocase) in both ventricles and reduced its microsomal expression in LV. After 60 min pacing they did not differ from the respective control values. Plasmalemmal expression of FATP-1 (fatty acid transport protein 1) increased and its microsomal expression decreased in RV after 30 min pacing. After 60 min pacing the plasmalemmal FATP-1 expression remained elevated whereas the microsomal expression did not differ from the control value. Pacing did not affect or expression of FABPpm (plasma membrane associated fatty acid binding protein) in either plasma membranes and microsomal compartments. Thirty min pacing increased plasmalemmal and reduced microsomal expression of GLUT-4 (glucotransporter 4) in both ventricles. It increased plasmalemmal expression of GLUT-1 (glucotransporter 1) in RV. It returned to normal after 60 min pacing. It is concluded that tachycardia induces numerous changes in metabolism of myocardial lipids as well as expression of fatty acid and glucose transporters in both heart ventricles.

Effect of atrial pacing on the level of bioactive sphingolipids in the heart ventricles of the rat.

Bioactive sphingolipids play important role in regulation of the function of the cardiomyocytes. There are no data available on metabolism of the sphingolipids in the heart under increased work-load produced by tachycardia. The aim of the present study was to examine effect of tachycardia on the level of the principal bioactive sphingolipids in the left and right ventricles. The experiments were carried out on male Wistar rats. After anesthesia, two electrodes were administered into the right common jugular vein so that their tips were placed at the vein's aperture. The resting heart rate was 355 ± 24/min and the rate of stimulation was 600/min. EKG was continuously monitored. The stimulation time was 30 and 60 min. Thereafter, blood from the abdominal aorta and samples of the left and right ventricle were taken. The following bioactive sphingolipids were quantified by means of high performance liquid chromatography: sphinganine, ceramide, sphingosine, sphingosine-1-phosphate and sphinganine-1-phosphate. In the left ventricle, 30 and 60 min tachycardia elevated the level of sphingosine, reduced the level of sphingosine-1-phosphate and sphinganine-1-phosphate. The level of ceramide was reduced only after 60 min. In the right ventricle, 60 min pacing resulted in elevation in the level of sphingosine and sphinganine and reduction in the level of other compounds studied. It is concluded that tachycardia induces changes in metabolism of bioactive sphingolipids in each ventricle. The changes may affect cardiomyocyte functions. Also, differences in sphingolipid metabolism between both ventricles are reported.

Consolidation of first-line therapy with busulphan and melphalan, and autologous stem cell rescue in children with Ewing's sarcoma.

According to the published report on current practice of hematopoietic SCT in Europe, high-dose therapy (HDT) with autologous stem cell support is a standard of care in paediatric patients with high risk (HR) or relapsed Ewing's sarcoma (ES). Randomized trials, however, have not confirmed the value of this procedure yet. In this retrospective analysis we intended to evaluate the role of HDT as a consolidation therapy in first remission of ES. A total of 102 patients were included in the analysis and divided according to the following risk factors: metastatic disease at presentation, feasibility of surgery and histological response after induction. Forty-one patients were classified as standard risk (SR) patients, while the remaining 61 children, with at least one risk factor, were classified as HR patients. HR group patients were non-randomized and qualified according to the decision of the local clinician to give a conventional consolidation (CC) or to perform high-dose chemotherapy and radiotherapy in selected patients. Twenty-six children were given CC while 35 patients were treated with HDT. The HDT consisted of oral BU 4 mg/kg p.o. in divided doses daily for 4 days (total dose 16 mg/kg) followed by melphalan 140 mg/m(2) i.v. on day -2. Probability of relapse-free survival (RFS) in median observation time was significantly worse in HR patients who were given CC therapy as compared with children with HR features receiving high-dose chemotherapy (0.27 vs 0.66 (P = 0.008); OS 0.31 vs 0.71 (P = 0.007), respectively). Patients from the SR group had a probability of RFS of 0.72 and OS of 0.75, and the difference between SR and HR patients after HDT was NS (P = 0.37). Our observation confirms that the consolidation of the first-line treatment with BU and melphalan improves the outcome in ES patients with HR features.

Changes in ECG and enzyme activity in rat heart after myocardial infarction: effect of TPP and MnCl2.

Heart infarction is one of the main causes of death in the human population. Assurance of a sufficient level of bioenergetic processes is very important for the heart after infarction. Mn2+ as well as thiamine pyrophosphate (TPP) are positive effectors of the pyruvate dehydrogenase complex (PDH) and the 2-oxoglutarate dehydrogenase complex (OGDH), both of which play a very important role in the Krebs cycle. Thus, we have established the effect of MnCl2 (10mg/kg) and TPP (20mg/kg)--4 injections every 12 h--on the activity of PDH, OGDH, lactate dehydrogenase (LDH) and malate dehydrogenase (MDH). Additionally, we perform an analysis of ECG to affirm the changes in the heart electrophysiology of healthy rats after MnCl2 and TPP treatment. We then analyzed changes in the activity of these enzymes after experimental myocardial infarction in rats. We observed a decrease of OGDH and MDH activity in rat hearts after infarction in comparison with sham-operated rats. Treatment of healthy rats with MnCl2 caused an increase of OGDH activity. Moreover both MnCl2 and TPP caused an increase of PDH activity and a decrease of MDH activity (TPP revealed a stronger effect). We found no changes in LDH activity. Electrocardiography data showed a slight shortening of the QT interval and an enhanced heartbeat rate after treatment with MnCl2. TPP caused only elongation of the QT interval. In conclusion, application of MnCl2 enhanced the activity of some very important enzymes in the respiration process (PDH and OGDH). This effect, connected with enhanced heartbeat and a slightly shortened ventricle relaxation, may have potential application during the key period of convalescence following heart infarction.

Changes in ECG and enzyme activity in rat heart after myocardial infarction: effect of TPP and MnCl2

Heart infarction is one of the main causes of death in the human population.Assurance of a sufficient level of bioenergetic processes is very important for theheart after infarction. Mn2+ as well as thiamine pyrophosphate (TPP) are positiveeffectors of the pyruvate dehydrogenase complex (PDH) and the 2-oxoglutaratedehydrogenase complex (OGDH), both of which play a very important role in theKrebs cycle. Thus, we have established the effect of MnCl2 (10mg/kg) and TPP(20mg/kg) - 4 injections every 12 h - on the activity of PDH, OGDH, lactate dehydrogenase(LDH) and malate dehydrogenase (MDH). Additionally, we perform ananalysis of ECG to affirm the changes in the heart electrophysiology of healthy ratsafter MnCl2 and TPP treatment. We then analyzed changes in the activity of theseenzymes after experimental myocardial infarction in rats. We observed a decrease ofOGDH and MDH activity in rat hearts after infarction in comparison with shamoperatedrats. Treatment of healthy rats with MnCl2 caused an increase of OGDHactivity. Moreover both MnCl2 and TPP caused an increase of PDH activity and adecrease of MDH activity (TPP revealed a stronger effect). We found no changes inLDH activity. Electrocardiography data showed a slight shortening of the QT intervaland an enhanced heartbeat rate after treatment with MnCl2. TPP caused onlyelongation of the QT interval. In conclusion, application of MnCl2 enhanced theactivity of some very important enzymes in the respiration process (PDH andOGDH). This effect, connected with enhanced heartbeat and a slightly shortenedventricle relaxation, may have potential application during the key period of convalescencefollowing heart infarction (AU)

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Neurologic complications in children after hemaopoietic stem cell transplantation: a single-center experience.

UNLABELLED: Neurologic complications may occur in patients undergoing haematopoietic stem cell transplantation (HSCT). The aim of the study was to evaluate the frequency and type of neurologic complications in children after HSCT. We performed a retrospective analysis of the incidence and outcome of neurologic complications among 171 consecutive children transplanted in one center. RESULTS: Among 84 autologous and 87 allogeneic (47 matched sibling donors, 31 matched unrelated donors, 8 mismatched family donors, and 1 cord blood) transplants, 7 patients (4%) developed neurologic complications, all of whom had undergone allogeneic transplantation (7/87 = 8%). These patients had relapses of acute leukemia (n = 3; acute myeloblastic in two and acute lymphoblastic in one), chronic leukemia, (n = 1), myelodysplastic syndrome (n = 2), and adrenoleudystrophy X (n = 1). Neurologic complications occurred after a median follow-up of 1 month (range, 14 days to 19 months). Of seven patients, four died. Neurologic complications were the cause in two patients. CONCLUSIONS: Among the analyzed material the risk of neurologic complications was lower than in other studies and these events were observed only in children undergoing allogeneic transplantation.

[Intertrochanteric fractures of the hip treated with dynamic hip screws (DHS)]. / Leczenie zlaman przezkretarzowych kosci udowej z uzyciem sruboplytki zeslizgowej (DHS).

The history and concept of sliding fixation is presented. Treatment results of 72 cases with 76 intertrochanteric fractures treated with DHS were analysed. Fractures were classified according to the Kyle scale into 4 different types. 61 males and 11 women age ranging from 46 to 72 years (average age: 65 years) were treated surgically. In 48 cases good results were achieved, in 22 cases results were satisfactory and in 6 cases results were poor. The use of DHS screws is an effective way of treating intertrochanteric fractures among patients with osteoporosis, as well as young patients requiring anatomic reduction of bone fragments. This technique allows early mobilization and weight bearing.

Our experiences in total condylar knee replacement.

The purpose of this study was to evaluate the outcomes of total knee replacement using endoprostheses. The material consisted of 78 knee endoprostheses in 68 patients operated in our Clinic over a 3-and-a-half-year period beginning in March 1995. Outcome assessment was based on clinical and radiological examinations before and after surgery, with due regard for the patient's subjective evaluation. The average follow-up period was 13 months. The majority of the patients were found to have improved walk efficiency, eliminated or reduced pain, improved mobility, corrected limb axis, and excellent subjective evaluation.

The application of screws covered with a layer of nanocrystaline diamond in tibia fractures treated with a Polfix fixator: further observations.

This article presents further results in the treatment of tibial shaft fractures using a Polfix fixator with screws covered with a layer of nanocrystaline diamond. The advantages of this new protective layer are presented in 16 additional patients. Research to date indicates that nanocrystaline diamond can be used successfully to cover the surface of surgical screws and plates.

[Characteristics of Lactobacillus strains contained in pharmaceuticals]. / Charakterystyka szczepów Lactobacillus wchodzacych w sklad preparatów farmaceutycznych.

The aim of the study was to characterize lactic acid bacteria (LAB) which are components of drugs administered orally in cases of intestinal disturbances, or antibiotic--related diarrhea. Biochemical properties, growth behavior, bile tolerance, and survival at low pH of six LAB strains (four strains L. rhamnosus and two L. acidophilus) were studied. The survival at low pH was determined in MRS broth (Difco) acidified to pH 1; 2; 3; and 4. Bile tolerance was tested on MRS broth with 0.3% oxgall (Difco). Between tested strains differences in ability to grow at low pH and survival in bile were observed. Only 0.01% inoculum of all examined strains survived at pH 1. Differences between strains in survival at low pH (pH 2 and pH 3) and tolerance of bile were observed. However, after 2 h incubation at pH 4, 100% of strains stayed alive. Examined strains demonstrated good 3% bile tolerance. All strains met the criteria for probiotic strains: ability to survive at pH 3 and in the presence of bile.

[Ender nails in the stabilization of trochanteric fractures of the femur in the elderly]. / Gwozdzie endera w stabilizacji zlaman przezkretarzowych kosci udowej u osób w podeszlym wieku.

Results of treatment for trochanteric fracture of the femur in 50 patients aged 25-91 (mean 71 years) are presented. In all cases reviewed the fracture has been stabilized with 3 or 4 Ender nails. In majority of cases bony union has been achieved. Short operating time, small incision remote from fracture site, early weight bearing constitute unquestionable advantages of the method. Most frequent complications as shortening of the extremity involved, excessive external rotation, pain related to migration of the nails, varus deformity at the fracture site were usually found in patients with marked osteoporotic changes.

Alcoholic typology and season of birth.

OBJECTIVE: This study analyzed the half years of birth of a large (n = 113,276) population of alcoholic patients in the U.S. Army Alcohol and Drug Abuse Prevention and Control Program from 1986 through 1990. METHOD: Subjects were enrolled for treatment of alcoholism or alcohol-related problems, and were analyzed for half year of birth. Groupings by age and gender, consistent with current theories of alcoholic typology, were compared, by means of chi-square tests, and by comparisons of two rates or proportions. RESULTS: The 17-21 year old and the 22-39 year old age groups differed by 5.1% in regard to half year of birth (chi-square = 260.317, p < 0.001, 95% CI:4.481 to 5.725, odds ratio: 1.23). Both groups differed significantly from the normal circannual birth pattern, but in opposite directions. CONCLUSIONS: The findings support the differentiation of types of alcoholics by age, which is a characteristic of Cloninger's classification, suggesting a biological, pre-natal factor.

Bone marrow transplantation does not ameliorate the neurologic symptoms in mice deficient in hypoxanthine guanine phosphoribosyl transferase (HPRT).

The use of bone marrow transplantation (BMT) for the treatment of genetic diseases with neurologic involvement has yielded mixed results. We have employed a mouse model of Lesch-Nyhan disease (LND) to assess the efficacy of BMT in ameliorating the neurologic manifestations of the disease. Adult HPRT-deficient mice exhibit a measurable decrease in striatal dopamine levels and a hypersensitivity to amphetamine. Marrow-ablated adult HPRT-deficient mice were transplanted with marrow from congenic HPRT-expressing mice. BMT altered neither the neurochemical nor the behavioral phenotypes in either HPRT-positive or HPRT-deficient mice. Barring any important species differences, these results suggest that BMT in its present form may not be an effective therapy for Lesch-Nyhan syndrome.

Influence of age and strain on striatal dopamine loss in a genetic mouse model of Lesch-Nyhan disease.

Lesch-Nyhan disease is a neurogenetic disorder caused by deficiency of the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT). Affected individuals exhibit a characteristic pattern of neurological and behavioral features attributable in part to dysfunction of basal ganglia dopamine systems. In the current studies, striatal dopamine loss was investigated in five different HPRT-deficient strains of mice carrying one of two different HPRT gene mutations. Caudoputamen dopamine concentrations were significantly reduced in all five of the strains, with deficits ranging from 50.7 to 61.1%. Mesolimbic dopamine was significantly reduced in only three of the five strains, with a range of 31.6-38.6%. The reduction of caudoputamen dopamine was age dependent, emerging between 4 and 12 weeks of age. Tyrosine hydroxylase and aromatic amino acid decarboxylase, two enzymes responsible for the synthesis of dopamine, were reduced by 22.4-37.3 and 22.2-43.1%, respectively. These results demonstrate that HPRT deficiency is strongly associated with a loss of basal ganglia dopamine. The magnitude of dopamine loss measurable is dependent on the genetic background of the mouse strain used, the basal ganglia subregion examined, and the age of the animals at assessment.

Effect of streptozotocin diabetes on fatty acid content and composition of the heart lipids in the rat.

The aim of the present study was to examine the effect of diabetes on long chain fatty acid content and composition in the heart lipids. The experiments were carried out on male Wistar rats. Diabetes was induced by intravenous administration of streptozotocin. Samples of the blood and the left ventricle were taken. Lipids were extracted and separated into different fractions. The following fractions were examined: free fatty acids, diacylglycerols, triacylglycerols and phospholipids. The fatty acids from each fraction were identified and quantified by means of gas-liquid chromatography. It was found that diabetes resulted in an almost four-fold elevation in the content of the free fatty acid fraction in the heart, whereas the plasma concentration of free fatty acids increased only two-fold. The diabetes induced changes in the content of particular acids in the fraction of free fatty acid in the heart did not reflect changes in their concentration in the plasma. The content of total di- and triacylglycerol fatty acids also markedly increased in diabetes. In both the compounds, the elevation in the content of individual acids, with the exception of myristic and palmitoleic acid reflected roughly the elevation in their concentration in the plasma. There were, however, several differences in the percentage composition of fatty acids between the two groups. In the fraction of phospholipids, the content of myristic, palmitic, stearic and linoleic acids remained stable, whilst the content of palmitoleic acid was reduced and the content of arachidonic acid was elevated. It is concluded that insulin deficiency results in marked changes in the endogenous lipid fatty acid content of the heart. These changes are not directly related to alterations in the supply of individual acids through the plasma.