Secondary structure inducing potential of beta-amino acids: torsion angle clustering facilitates comparison and analysis of the conformation during MD trajectories.

While numerous examples of beta-peptides--exclusively composed of beta-amino acids--have been investigated during the past decade, there are only few reports on the conformational preference of a single beta-amino acid when incorporated into a cyclopeptide. The conformational bias of beta-amino acids on the secondary structure of cyclopeptides has been investigated by NMR spectroscopy in combination with distance geometry (DG) and molecular dynamics (MD) calculations using experimental constraints. The atomic coordinate RMSD criterion usually employed for clustering of conformations after DG and MD calculations does not necessarily group similar peptide conformations, as there is an insufficient correlation between atomic coordinates and torsion angles. To improve on this shortcoming and to eliminate any arbitrary decisions during this process, a torsion angle clustering procedure has been implemented. For the cyclic pentapeptides cyclo-(-Val-beta-Hala-Phe-Leu-Ile-) 1 and cyclo-(-Ser-Pro-Leu-beta-Hasn-Asp-) 3, the beta-amino acid is found in the central position of an extended gamma-turn (pseudo gamma-turn, Psigamma-turn), while the beta-Hpro residue in the cyclic hexapeptide cyclo-(-Ser-beta-Hpro-Leu-Asn-Ile-Asp-) 5 preferentially occupies position i+1 of a pseudo beta-turn (Psibeta-turn). These results further corroborate the hypothesis of beta-amino acids being reliable inducers of secondary structure in cyclic penta- and hexapeptides. They can be employed in the de novo design of biologically active cyclopeptides in pharmaceutical research, since the three-dimensional presentation of pharmacophoric groups in the side chains can be tailored by incorporation of beta-amino acids in strategic sequential positions.

[Early diagnosis, therapy, follow-up and survival in patients with thyroid malignancies]. / Rana dijagnostika, terapija, pracenje i prezivljavanje bolesnika sa malignitetom stitaste zlezde.

UNLABELLED: The Aim of the paper was to give a review of an early diagnosis, therapy, follow-up and survival rate of patients with thyroid malignancy (TM). The paper presented the algorithm of early diagnosis: clinical, scintigraphic and ultrasonographic examination together with fine needle biopsy, cytologic analysis of the smear and biopsy ex tempore of the clear and suspected thyroid node to malignancy. Therapy of all TM forms was mainly surgical: postsurgical treatment was dependent on the type of malignancy: radioiodine 131-I, radiologic treatment, chemotherapy and radioimmunotherapy, (the latest one being in the phase of a clinical research). Follow-up was in accordance with the protocol and it was necessary because it contributes to the survival rate. In the presentation of survival rate for differentiated and medullar carcinomas we gave our results and literature data, while for the other malignancies only data from literature were presented. CONCLUSION: Only an early diagnosis of the nodular goiter together with an up-to-date treatment can cure TM patients in a high percentage and prevent development of a terminal stage of the disease which is extremely severe in all forms of this malignancy.

[131-I-lipiodol in therapy of liver carcinoma--methods and case report]. / 131-J-lipiodol u lecenju karcinoma jetre--metodologija i prikaz slucaja.

INTRODUCTION: Primary malignant liver tumors are successfully treated only by means of surgery, but no more than 10% of patients are candidates for surgical intervention. The rest receive only palliative treatment which is, as a rule, unsuccessful. 131I-Lipiodol therapy (commercial label LIPIOCISTM) applied to the hepatic artery through a catheter has been used since 1984, primarily for treatment of hepatocellular carcinoma (HCC). The aim of this paper is to review the methodology of 131J-Lipiodol treatment of primary liver carcinomas. This treatment was applied for the first time in Yugoslavia. CASE REPORT: A female patient, 46 years of age had an inoperable primary liver carcinoma. Since progression of the disease couldn't be controlled by chemotherapy, treatment with 131I-Lipiodol was indicated. After blocking the thyroid with Lugol, Sol. 2.22 GBq of 131I-Lipiodol was injected into hepatic artery via catheter, and the patient was isolated in a designated facility until discharged. Around 5 months later, second therapeutic dose of 1.11 GBq was administered. Early post-therapy complications were severe, but transient. After 131I-Lipiodol therapy, the tumor growth was stopped, but the patient's general condition slowly deteriorated. The patient died 7 months after receiving the first therapeutic dose. CONCLUSION: In the reported patient, 131I-Lipiodol therapy stopped the tumor growth within the liver and significantly prolonged survival compared to the expected, but no improvement in quality of life was achieved. This treatment methodology is very complex. Medical staff providing care for these patients is exposed to substantial irradiation.

[Detection of K-ras and p-53 oncogene mutations in a patient with acute lymphoblastic leukemia before and after bone marrow transplantation]. / Detekcija mutacija u K-ras i p-53 onkogenima kod obolelog od akutne limfoblastne leukemije pre i posle transplanticije kostine srzi.

We present a case of 22 years old male patient, who was submitted to singenic transplantation of hematopoietic cells originating from the bone marrow in the remission phase of the diagnosed acute lymphoblastic leukemia (ALL). The bone marrow sample was donated by his healthy twin brother. The pretransplantation and transplantation phases were regular. We analyzed the presence of K-ras and p-53 point mutations in our patient with ALL and for the first time we had the opportunity to analyze the samples from two monozygotic twins. DNA was isolated from the peripheral blood mononuclear cells (PBMNC) by the standard procedure, of the patient with ALL before and after bone marrow (BM) transplantation and of his clinically healthy twin brother. Samples were subjected to PCR amplification of K-ras exons 1 and 2 and p-53 exons 5, 6, 7 and 8. In PBMNC of the patient with ALL before BM transplantation, mutations were observed in exon 1 of K-ras and exon 8 of p-53 gene. These mutations were found neither in PBMNC sample of his twin brother, nor in PBMNC of the patient with ALL after BM transplantation. In the p-53 exons 5, 6 and 7 and exon 2 of K-ras, there were no mutations in any analyzed samples. Detected mutations in K-ras and p-53 genes could be a part of larger genetic abnormalities and the obtained results had shown the possibility of using DNA mutational changes in the follow-up of the success of BM transplantation. The molecular disease marker that was found by this method was also significant for the detection of minimal residual disease at the molecular level.

2,8-Dihydroxy-1,3,7,9-tetramethyl-6,12-dihydrodipyrido

The 2,8-dihydroxy-1,3,7,9-tetramethyl-6,12-dihydrodipyrido[1,2-a:1', 2'-d]pyrazinediylium dication possesses 2/m symmetry and lies in the mirror plane together with a chloride anion and the water O atom. The dication also lies on an inversion centre, i.e. C(16)H(20)N(2)O(2)(2+).2Cl(-).2H(2)O. Due to these symmetry constrictions the dication adopts an unexpected planar conformation. Molecules are linked by O-H.O and O-H.Cl hydrogen bonds to form chains, which are cross-connected by C-H.Cl attractive interactions forming a complex three-dimensional hydrogen-bond network.

[Treatment of Hodgkin's disease with high-dose chemotherapy and autologous stem cell transplantation]. / Lecenje Hodgkin-ove bolesti visokim dozama hemioterapeutike uz autologu transplantaciju maticnih celija hematopoeze.

High dose chemotherapy (HDC) with autologous stem cell transplantation (SCT) was applied for the treatment of 13 patients (pts) with Hodgkin's disease (HD) (10 with relapsed form and 3 with conventional chemotherapy resistant form) in the Clinic for Hematology, Military Medical Academy, from May 1997 to October 1999. After the initial treatment for the reduction of tumor, burden stem cells were mobilized by cyclophosphamide 2.5-7.0 g/m2 with G-CSF 5-12 micrograms/kg body mass (BM). The average number of colected mobilized mononuclear cells (MNC) was 2.99 (1.66-5.9) x 10(8)/kg BM by the apheresis large volume from peripheral blood. All patients received BEAM (BCNU, etoposide, Cyto-Ara, and melfalan) conditioning regimen with adequate supportive therapy. Good engraftment (100%) was observed at postransplantation period: number of polymorphonuclear cells was > 0.5 x 10(9)/l, on day 13th (10-21) and number of platelets > 20 x 10(9)/l, on day 17th (11-28). One patient (7.6%) died due to infective complications at day 98th after transplantation, 9 (69.2%) patients achieved complete and 3 (23.1%) patients partial remission of the disease. Out of three patients with partial remission, one relapsed, seven months after autologous SCT, with conventional chemotherapy resistant form and two, after the applied conventional locoregional radiotherapy reached remission. One patient (7.6%) developed secondary malignancy of acute myeloid leukemia form with threelinage displasy 27 months after autologous SCT. HDC with autologous SCT contributes to more successful treatment of early relapsed and standard chemotherapy resistant forms of HD and gives the opportunity for successful quality of living for those patients.

Donor leukocyte infusion--the effect of mutual reactivity of donor's and recipient's peripheral blood mononuclear cells on hematopoietic progenitor cells growth.

Donor leukocyte infusions are an effective therapy for patients who relapse with leukemia after bone marrow transplantation. We report the case of 14-year-old boy who relapsed 34 months after sibling donor bone marrow transplant for Philadelphia-positive chronic myeloid leukemia. Subsequently, he received three infusions of donor mononuclear cells (DMNC) harvested in steady state hematopoiesis and one G-CSF mobilized-peripheral blood mononuclear cells (PBMC) infusion. Simultaneously, test named as--"Test of Mixed Progenitors" (TMP) was performed for the assessment whether the outcome of donor leukocyte infusion treatment could be predicted. Prior to DMNC infusions, the CFU-GM and BFU-E colony assays were performed for donor's and recipient's PBMC individually, as well as for the mixture of these cells at 1:1 ratio. The cells were plated either directly in the semisolid medium or after 24 h preincubation treatment. Significantly lower values for CFU-GM derived colonies were determined in TMP in comparison to the CFU-GM values obtained for the recipient's cells. The reduced number of CFU-GM was determined both in TMP performed without preincubation treatment, app. 80% and after the 24 h preincubation, app. 55%. The reduced number of BFU-E derived colonies (app. 44%) was observed only related to recipient's cells and after the preincubation treatment of the cells. The patient did not develop GVHD and currently (40 months after the first infusion). He remained well in complete hematological, cytogenetic, molecular and clinical remission, which was the most direct evidence of the GVL effect. The novel in vitro TMP test in which the specific contribution of donor's leukocytes to the growth of recipient's hematopoietic precursor cell growth was determined, correlated with the clinical outcome.

[Secondary myelodysplastic syndrome after autologous transplantation of hematopoietic stem cells in a female patient with Hodgkin's disease]. / Sekundarnih mijelodisplazni sindrom nakon autologe transplantacije maticnih celija hematopoeze kod bolesnice sa Hodgkin-ovom bolesti.

Secondary acute myeloid leukemia with threelineage displasy (sAML/MDS) has been described as a complication of therapeutic approach with high-dose chemotherapy and autologous stem cell transplantation (SCT) in the patients with Hodgkin's disease (HD). It is not yet clear whether the sAML/MDS is a consequence of a standard therapeutic regimen, applied before transplantation, or a high-dose chemotherapy. In a female patient with initially resistant form of HD at III-B-b clinical stadium (bulky disease in neck and mediastinum) after the initial treatment (MOPPx4; ABVDx3; BEA-COPPx2), high-dose chemotherapy has been applied according to BEAM protocol with autologous SCT. In a period after transplantation, radiotherapy (RT) has been applied at initial region of the disease and a patient reached complete remission (CR), which lasted for a 27 months. After that period sAML/MDS has been observed. Application of more standard therapeutic cycles and characteristic cytogenetic findings are the facts that support the opinion that sAML/MDS is a consequence of standard treatment before transplantation, rather than high-dose chemotherapy. That finding implies the need for correct choice of the HD patients suitable for early SCT therapeutic approach.

[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.

[In vivo study of spontaneous and therapy-induced apoptosis in patients with chronic lymphocytic leukemia treated with chlorambucil]. / In vivo pracenje spontane i terapijom indukovane apoptoze kod bolesnika sa HLL lecenih hlorambucilom.

The efficiency of Chlorambucil in the induction of apoptosis was investigated in the study, and measurable apoptosis parameters were compared to the other prognostic factors with the aim of possible prediction of clinical response to the therapy in the patients with CD5 + B-cell chronic lymphocytic leukemia (B-CLL). Seven newly diagnosed patients, initially treated with daily high-doses of Chlorambucil (HD-CLB) were analyzed. Quantitative analysis of apoptosis parameters on semi-fine sections obtained from peripheral blood was performed prior and during the first five days of therapy. The level of spontaneous apoptosis (SA) was determined, as well as the maximal response by apoptosis (MAR), and the time needed to establish maximal response by apoptosis (TMAR), respectively. The results revealed that the level of SA in the studied group of patients was 11.39%-20.50%. In three patients with achieved criteria for complete remission (CR) was observed high level of SA, TMAR 2-4 days and MAR 23.42-26.36%, respectively. All patients with CR were with negative LDT, non-diffuse involvement of bone marrow and clinical stage B. Criteria for partial remission (PR) were achieved in 4 patients. Within this group, all three measurable parameters of apoptosis could have been determined in only one patient, while in the rest was noticed the increased percentage of apoptotic cells on the last day of follow-up. In all patients was observed negative LDT, diffuse bone marrow involvement, and 2 out of 4 patients had CLPL of cytomorphological type and clinical stage B. By comparing the obtained values of measurable apoptotic parameters with the clinical response to the applied therapy with HD-CLB, it is possible to divide our patients into two groups: patients who have achieved CR have the highest percentage of cells dying due to the therapy-induced apoptosis, as well as the higher values of measurable parameters compared to the certain parameters of the patients with the criteria for PR. Our preliminary results of therapeutic response to the apoptosis might be useful for the timely decision upon the duration of therapy and change of modality of treatment for every patient during the follow-up period.

Evaluation of cryopreserved murine and human hematopoietic stem and progenitor cells designated for transplantation.

The influence of five different cryopreservation protocols on the quality and/or quantity of frozen cells was investigated on mouse bone marrow cells and human peripheral blood mononuclear cells (MNC). The efficiency of the protocols was evaluated on the basis of the recovery of very primitive pluripotent hematopoietic stem cells (MRA), pluripotent progenitors (CFU-Sd12), committed granulocyte-monocyte progenitors (CFU-GM) of mouse cells after thawing. The recovery of MRA, CFU-Sd12 and CFU-GM varied depending on the type of freezing procedure and cryoprotector (DMSO) concentrations used. It was shown that the controlled-rate protocol was more efficient, enabling better recovery of all categories of progenitor cells in frozen samples. The most efficient was the controlled-rate protocol of the cryopreservation designed to compensate for the release of fusion heat, which enabled the best recovery of CFU-GM (73.0 +/- 8.8%) and CFU-Sd12 (90.0 +/- 15.9%) when combined with 5% DMSO concentration (protocol 4). On the contrary, a better recovery (79.8 +/- 13.5%) of very primitive stem cells (MRA) was achieved only when the higher concentration (10%) DMSO was used in combination with a five-step protocol of cryopreservation (protocol 1). These results pointed out the adequately used controlled-rate freezing to be essential for a highly efficient cryopreservation of some of the categories of hematopoietic stem and progenitor cells. At the same time, it was obvious that a higher DMSO concentration was necessary for the cryopreservation of MRA, but not for more mature progenitor cells (CFU-S, CFU-GM). These results imply the existence of a mechanism that decreases the intracellular concentration of DMSO in MRA cells, which is not the case in less primitive progenitors. For human MNC, the recovery and viability of the cells, as well as the engraftment potential of cryopreserved cells after thawing were investigated. Cryopreservation protocol 1 resulted in better MNC recovery (82.7 +/- 10.4%) than protocol 3 (49.9 +/- 15.1%). The mean recovery of MNCs (collected from patients for autologous transplantation) was 78.5 +/- 7.3% (protocol 1) and 53.1 +/- 26.2% (protocol 3). The obtained favorable recovery of thawed cells and rapid reconstitution of hematopoiesis (on the day 11th following the transplantation) in patients confirmed that the controlled-rate freezing in combination with optimal DMSO concentration was able to obtain sufficient progenitor cryoprotection.

[Neurologic disorders in a patient with autoimmune hemolytic anemia]. / Neuroloski poremecaji kod bolesnika sa autimunskom hemolitickom anemijom.

Completely different entities might be with the same possibility in the baseline of interweaving of symptoms and signs of nervous system damage. One of them, the deficiency of vitamin B12 very frequently causes megaloblastic anemia and funicular myelosis. In the case of our patient, after the clinical picture of hemolytic anemia was revealed, by slow-progressive course was developed neurologic deficiency that, according to its features, could have the deficiency of cobalamin and folic acid in its etiologic background. On the basis of disease course, clinical finding, numerous clinical investigations so as the reaction to applied therapy it was assumed that the patient had besides confirmed autoimmune hemolytic anemia the pernicious anemia as the associated cause of anemic syndrome and the basic reason of the development of neurologic deficiency. Described is the frequent associated occurrence of pernicious anemia and antiglobulin positive hemolytic anemia, so as the significant association of pernicious anemia with the deficiency of immunoglobulins that was otherwise observed in our patient as the permanent IgA deficiency.

Mass spectral fragmentation patterns of some new 3,7-dichloro-benzo[1,2-b:4,5-b']dithiophene-2,6-dicarboxylic acid dianilides and 3,5-dichloro-dithieno[3,2-b:2',3'-d]thiophene-2,6-dicarboxylic acid dianilides. II.

The electron impact mass spectra of some benzo[1,2-b:4,5-b']dithiophene-2,6-dicarboxylic acid dianilides and dithieno[3,2-b:2',3'-d]thiophene-2,6-dicarboxylic acid dianilides are discussed. Dominant peaks in these dianilides are formed by the cleavage of a C-N bond on one side of an anilino group. These ions fragment further by the cleavage of a C-C bond on the other side of an anilino group and a CONRPhR' group may be lost directly. After loss of CO, the characteristic benzodithiophene radical cation, C10H2S2Cl2[symbol: see text], at m/z 256 and the dithienothiophene radical cation, C8S3Cl2[symbol: see text], at m/z 262 are formed from their respective precursor compounds.

[Use and efficacy of radioiodine therapy in the treatment of hyperthyroidism]. / Primena i efikasnost radiojodne terapije kao nacin lecenja hipertireoze.

Radioiodine (131J) therapy is a method of radical treatment for hyperthyroidism. In our study in the period 1971-1993, we administered radioiodine therapy in 163 patients with hyperthyroidism. We performed a long-term follow-up, from one month, to 21 years after the radioiodine therapy. The cured rate was 83.4%: euthyroid state was found in 43.6% of patients and hypothyroidism appeared in 39.9% of patients. After the radioiodine therapy hyperthyroidism was found in 16.6% of cases. Radioiodine therapy is very efficient, non-invasive, radical treatment of hyperthyroidism, not expensive and easy to administer. It has practically no immediate or long-term complications, except hypothyroidism.

Protocol of the diagnostic, therapy and follow-up of differentiated thyroid cancer.

The Protocol of the multidisciplinary diagnostic treatment and follow-up of differentiated thyroid cancers, made in the Institute of Oncology in Sremska Kamenica, is described. It is in use in this institution from 1990. It is in concordance with general oncology principles and follows the guidelines of the International Union Against Cancer (UICC) concerning this problem.

[Antilymphocyte globulin in the treatment of aplastic anemia]. / Antilimfocitni globulin u lecenju aplasticnih anemija.

Between January 1986 and July 1990, 17 patients with acquired aplastic anemia were treated with ALG or ATG combined with high doses of methylprednisolone. The mean age was 24.3 years (from 4 to 51 years). There were 9 cases with idiopathic etiology of acquired aplastic anemia; in 7 cases aplastic anemia was developed during or after HBsAg infection. In one case aplastic anemia was developed during tuberculous kidney infection. The remission of the disease was achieved in 11 of 17 cases (complete remission in 9-53%, and partial in 2-12% patients). Six (35%) patients did not respond to the treatment with ALG. One patient died of infection and hemorrhagic complications, two weeks after the therapy, without responding to the treatment with ALG. The four year survival rate without recidives was 65% (11/17). Four (23.5%) patients developed clonal diseases: PNH in 2; MDS in 1 and AL in 1 patient, 24, 38, 9 and 6 months after the therapy with ALG, respectively. The age of the patients is a valuable prognostic parameter (all patients under 20 years of age entered the remission), which cannot be said for pretreatment levels of reticulocytes, neutrophils and platelets. In none of the patients adverse effects of ALG were observed. The treatment was conducted in isolated rooms with sterile air circulation. ALG combined with high doses of methylprednisolone, for the majority of patients with aplastic anemia represents a drug of choice and is an appropriate alternative therapy to alogenic bone marrow transplantation, especially for patients with no HLA identical bone marrow donor.

[Dosimetric measurements and early complications in patients with differentiated thyroid carcinomas treated with radioactive iodine]. / Dozimetrijska merenja i rane komplikacije kod bolesnika sa diferenciranim karcinomom stitaste zlezde lecenih radioaktivnim jodom.

The aim of the study was to determine the exposure dose rate during the application of radioiodine therapy (ablative or tumoral dose) given in order to treat the differentiated thyroid carcinoma, during the medical visit and examinations of those patients, to establish the safety distance from patients both for population and for medical staff and to perceive early complications after the therapy. The dosimetric measurements were performed in 10 patients. The exposure dose rate during the application of the therapy ranged from 2000 to 10000 pC/kg.s, during the visit from 528 to 15 pC/kg.s and during the examinations of patients from 5500 to 200 pC/kg.s. The average safety distance from patients for population was about 8.5 m on the day O (the very day of the therapy) and 2.0 m on the day 4, while for the medical staff it amounted to 5.0 m on the day 0 and 0.5 m on the day 4. The early complications perceived were as follows: radiation thyroiditis in 5/10 patients, stomach problems in 1/10 patients and transitorial leucopeny, forty days after the therapy, in 2/10 patients.