Frequency of BCR-ABL fusion transcripts in Serbian patients with chronic myeloid leukemia.

PURPOSE: The aim of this study was to analyze the occurrence of the most frequent BCR-ABL transcript variants (b3a2, b2a2 and e1a2) in Serbian patients with chronic myeloid leukemia (CML) and compare it with the occurrence reported in other populations. METHODS: We analyzed peripheral blood and bone marrow samples of 136 Serbian patients with CML by RT-PCR and cytogenetic methods. RESULTS: In 100 patients (73.5%) the b3a2 and in 34 (25%) the b2a2 forms of BCR-ABL were detected. One (0.75%) patient was BCR-ABL negative, but in lymphoblastic transformation he expressed the e1a2 [corrected] transcript of BCR-ABL. One (0.75%) patient displayed both b2a2 and b3a2 forms of BCR-ABL. Analysis of this group according to karyotype showed b3a2 predominance (79%) in patients with classic t(9;22); b2a2 was found in 20% and both b2a2 and b3a2 forms in 1%. In variant translocations b3a2 in 65% and b2a2 in 35% of the patients were detected. In contrast, the subgroup with normal karyotype expressed slight predominance of the b2a2 form (50%); b3a2 was found in 43% of the patients and one patient (7%) displayed e1a2. CONCLUSION: Predominance of the b3a2 form in Serbian patients with CML is in concordance with other relevant investigations, conducted mostly on Caucasian ethnic groups, but in contrast to the study performed on the Mestizo ethnic group in Ecuador. Slight predominance of the b2a2 form was also noticed among the patients with normal karyotype.

Alternative respiration of fungus Phycomyces blakesleeanus.

Respiratory characteristics of germinating spores, developing mycelium and mitochondria of the fungus Phycomyces blakesleeanus were investigated by means of oxygen Clark-type electrode. The effects of respiratory inhibitors and metabolic compounds on oxygen consumption were tested. It was demonstrated that P. blakesleeanus apart of cyanide-sensitive respiration, CSR, possess alternative respiration, (cyanide-resistant respiration, CRR) which is constitutive and whose capacity decreases during development. Maximum is observed for activated spores where CRR capacity is significantly greater than CSR. After treatment with antimycin A, a third type of respiration insensitive to antimycin A and low concentration of SHAM (sufficient for inhibition of CRR), but sensitive to cyanide and high concentration of SHAM, has been expressed.

Expression of programmed cell death proteins in patients with chronic myeloid leukemia.

PURPOSE: Chronic myelogenous leukemia (CML) is a malignant myeloproliferative disease developing out of pluripotent hematopoietic stem cells that contain the fusion Bcr-Abl gene. The mechanisms that lead to these changes at molecular level are still unknown as are the mechanisms that increase the proliferative capacity of these cells. Disorders that occur in the process of apoptosis represent one of the possible molecular mechanisms that bring about disease progress. In our study we analyzed the presence of mutated (mut) p53 gene and the amplification of Bax proteins in patients with CML. PATIENTS AND METHODS: This study included 30 patients with CML (23 in chronic phase, 7 in blast transformation). Using immunohistochemistry with alkaline phosphatase / anti-alkaline phosphatase (APAAP) method we analyzed the expression of cell death proteins p53 and Bax in mononuclear bone marrow cells. Polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) method was used to analyze the presence of mut p53 gene in mononuclear peripheral blood cells. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to analyze the presence of Bcr-Abl in peripheral blood cells. RESULTS: High expression of Bax protein was detected in all analyzed patients, but no significant differences were noticed among them. No mut p53 gene was detected in any of the analyzed samples. Bcr-Abl b3a2 protein form was detected in all patients with variant translocations. CONCLUSION: Lack of mut p53 product in the peripheral blood and bone marrow cells in patients with CML suggests that this gene plays no important role in disease pathology. Increased level of Bax protein expression is an essential characteristic of CML cells but it is not related with the clinical stage of disease.

Ventricular and vascular remodelling effects of the angiotensin II receptor blocker telmisartan and/or the angiotensin-converting enzyme inhibitor ramipril in hypertensive patients.

Angiotensin II induces inflammatory activation of vascular smooth muscle cells and can cause left ventricular hypertrophy (LVH). Telmisartan is an angiotensin II receptor blocker with demonstrated beneficial effects on cardiac and vascular structure and function in animal models. The angiotensin-converting enzyme inhibitor ramipril also reduces ventricular and vascular remodelling. The open-label study observed 75 treatment-naive, moderately or severely hypertensive (systolic blood pressure 160-190 mmHg, diastolic blood pressure 90-110 mmHg) patients (age range, 42-58 years) treated with once-daily telmisartan 40 mg force-titrated to 80 mg after 1 month (n=25), once-daily ramipril 2.5 mg force-titrated to 5 mg after 1 month (n=25), or once-daily telmisartan 40 mg plus ramipril 2.5 mg (n=25); the total duration of treatment was 6 months. At baseline, blood pressure, left ventricular mass index (LVMI), carotid intima-media thickness (IMT) and carotid cross-sectional intima-media area (CSA) were measured. Measurements were repeated 1, 3 and 6 months after initiation of treatment. After 6 months, comparable blood pressure reductions were achieved with the three treatments. Reductions in LVMI after 6 months' treatment were 11.4%, 9.9% and 15.6% with telmisartan, ramipril, and telmisartan plus ramipril, respectively. Respective reductions in IMT were 14.6%, 12.0% and 18.2%, and for CSA were 7.8%, 4.3% and 11.5%. In conclusion, treatment with telmisartan or ramipril for 6 months resulted in regression of LVH and vascular remodelling. When a combination of telmisartan and ramipril was administered, additional regression and remodelling occurred.

Light-induced transient ion flux responses from maize leaves and their association with leaf growth and photosynthesis.

Net fluxes of H+, K+ and Ca2+ ions from maize (Zea mays L.) isolated leaf segments were measured non-invasively using ion-selective vibrating microelectrodes (the MIFE technique). Leaf segments were isolated from the blade base, containing actively elongating cells (basal segments), and from non-growing tip regions (tip segments). Ion fluxes were measured in response to bright white light (2600 micromoles m-2 s-1) from either the leaf segments or the underlying mesophyll (after stripping the epidermis). Fluxes measured from the mesophyll showed no significant difference between basal and tip regions. In leaf segments (epidermis attached), light-induced flux kinetics of all ions measured (H+, Ca2+ and K+) were strikingly different between the two regions. It appears that epidermal K+ fluxes are required to drive leaf expansion growth, whereas in the mesophyll light-induced K+ flux changes are likely to play a charge balancing role. Light-stimulated Ca2+ influx was not directly attributable either to leaf photosynthetic performance or to leaf expansion growth. It is concluded that light-induced ion flux changes are associated with both leaf growth and photosynthesis.

[Second allogenic bone marrow transplantation after late graft rejection in a patient with severe aplastic anemia]. / Sekundarna alogena transplantacija kostne srzi kod bolesnika sa teskom aplastickom anemijom nakon kasnog odbacivanja primarnog kalema.

Allogeneic bone marrow transplantation (BMT) is the treatment of choice in young patients (pts) with severe aplastic anemia (SAA) who have an HLA identical sibling donor. Late graft rejection to following allogeneic BMT for SAA is a significant clinical problem and is associated with a high risk of mortality. The optimal treatment strategy for patients with late graft rejection after first BMT is still an open question. We report 12-year-old patient with acquired SAA who underwent BMT from his HLA identical sister. BMT was first-line treatment within 3 months of diagnosis. Preparative therapy was Cyclophosphamide (Cy) 200 mg/kg body mass (BM) during 4 days. Graft versus host disease (GVHD) was prevented with Methotrexate (MTX), Methylprednisolone (MPDN) and Cyclosporin A (CsA). After 17 months, during which patient was with normal blood counts and full donor chimaerism, graft rejection occurred. The patient was re-engrafted from the same donor after conditioning with Cy 200 mg/kg BM plus horse antithymocyte globulin (ATG)--2 vials (á 25 mg)/10 kg BM over 4 days. Before the collection, donor's bone marrow was activated with low dose rhGM-CSF (3 micrograms/kg one day). Following a secondary BMT engraftment has sustained. The patient is alive with full donor chimaerism 26 months from secondary transplantation, without acute or chronic GVHD.

[Diseases of the upper segment of the gastrointestinal tract and Helicobacter pylori infection]. / Oboljenja gornjih segmenata gastrointestinalnog trakta i infekcija Helicobacterom pylori.

Helicobacter pylori infection is the most frequent infection in humans. It is assumed that over 50% of total world population suffer from this infection. Chronic gastritis is the most usual consequence of this infection. Development of peptic ulcer appears in case of 10 to 20 percent of infected people with the increased risk of malignancy. In the group of 819 examined patients at the Clinic for gastroenterology and hepatology of the Institute for internal diseases, the presence of helicobacter pylori was confirmed in case of 69.18% of patients. The infection was equally observed in both sexes and its frequency was higher with the increase of age, reaching its peak in the sixth decade of life (75.24%). The highest frequency was found in patients with duodenal ulcer (84.85%) and hyperplastic stomach polyp (81.82%) and lowest in case of resected stomach (56.52%). We also establish the dependence of the type and degree of pathohistological changes and the Helicobacter pylori infection.

[Ixodes ricinus proven as a vector of Lyme borreliosis in Yugoslavia]. / Ixodes ricinus dokazan vektor lajm borelioze i u Jugoslaviji.

263 fasting female Ixodes ricinus were examined for Borrelia burgorferi, the vector of Lyme borreliosis. Female ticks were collected by flagella in the biotopes (Belgrade and Osijek) in which ticks bite patients with Lyme borreliosis. Borrelia burgorferi was proved in 58 (22%) of 263 female ticks in the native preparations of the intestinal contents by darkfield microscopy. In macerates of two groups of 5 female ticks each, Borrelia burgorferi was isolated by cultivation in the modified Kelly nutrient media for borrelia. The supposition that Ixodes ricinus is the vector of Lyme borreliosis in Yugoslavia is confirmed.

[Risk of infection with Borrelia burgdorferi in an area endemic for Lyme disease]. / Rizik od infekcije borelijom burgdorferi u endemskom podrucju lajmske bolesti.

The results of the study of the prevalence of antibodies against Borrelia burgdorferi in three categories of subjects from the endemic region of Lyme disease in the northeast Croatia are reported. The study comprised 265 subjects: 97 living in the rural region, 51 forest rangers working in this region and 117 members of the Yugoslav People's Army who spent a certain period of time in this region. Positive titer of specific antibodies were found in 11.3% of local inhabitants, 25.5% forest rangers and 6% soldiers. Serologic testing was performed by the direct immunofluorescent method with aspiration and the titer in the dilution of 1:80 was considered significant. Some risk factors causing disease as well as the relationship between inapparent and clinically manifested cases of infection are pointed out.

[Behavior of Zn-phosphate cement in parafunctional conditions. 2. Wear resistance of Zn-phosphate cement]. / Ponasanje Zn-fosfat cementa u uslovima parafunkcionih aktivnosti. II deo: Otpornost na habanje Zn-fosfat cementa.

The paper deals with the phenomenon of complex stress of the Zn-phosphate cement when slope specimens are loaded. In a 5 degree slope the horizontal component was only 0.7% whereas this value began to increase up to 45.3% when the slope was 40 degrees. It is of interest that in the slant specimens the significant part of the total force referred to horizontal component, particularly expressed in parafunctions, which speaks for the unfavourable stress type suffered by specimen slope. The second part of the experiment revealed the degree of wear of Zn-phosphate cement parafunctional activities. The value of 1800 which presents the number of contacts realized by a patient with parafunctions only in the course of daily action of occlusive dental surfaces, was selected as the maximal number of pulses. The following was established: with the increase of pulses the degree of wear was also increased; the wear was more expressed in the mobile specimen; the degree of wear of the mobile specimen was not a linear function of the number of pulses contrary to the fixed one in which a linear dependence was observed.