OC-07 - Decoding risk for thromboembolic events in lymphoma patients.

INTRODUCTION: There are few prediction tools for estimating the risk of thrombosis but they are based on studies performed on hospitalized medical patients without cancer or on hospitalized neutropenic cancer patients without special consideration to lymphoma patients. AIM: Aim of our study was to determine incidence of thromboembolic (TE) events in patients with non Hodgkin lymphoma (NHL), Hodgkin lymphoma (HL) and chronic lymphocytic leukemia (CLL)/ small lymphocytic lymphoma (SLL) who were hospitalized to the lymphoma department in the Clinic of hematology, Clinical Center Serbia, Belgrade and Clinic of hematology, Clinical Center Kragujevac. Also, we assessed 2 predictive models (Padua and Khorana score) and create new model for the identification of lymphoma patients at risk for thromboembolism. MATERIALS AND METHODS: We reviewed all medical records of patients with with NHL, HL and CLL/SLL diagnosed and treated at two previously mentioned institution between January 2006 and December 2014. RESULTS: The study population included 1820 eligible lymphoma patients. Of all the patients included in the study, 99 (5.4%) developed at least one TE during a follow-up period of 3 months from the end of therapy. In the final multivariate analysis, the following variables were independently associated with risk of TE: previous VTE and/or arterial events, reduced mobility (ECOG 2-4), obesity (BMI >30 kg/m(2)), extranodal localization, mediastinum involvement, development of neutropenia during therapy and hemoglobin level less than 100g/L. Subsequently, we assigned points for the risk model based on the regression coefficients obtained from the final model and developed Thrombosis Lymphoma (ThroLy) score consisting of all significant variables from the multivariate analysis. The Throly score was arrived at by assigning 2 points for all parameters with an OR >5 in multivariate regression analyses (e.g., previous VTE and arterial events, mediastinum involvement, and BMI) and 1 point for rest all other significant variables. Finally, population were divided into 3 risk categories for TE based on the score from the risk model: low (score 0-1), intermediate (score 2-3) and high (score >3). High risk score had a positive predictive value (probability of TE in those designated high risk) of 65.2%. CONCLUSIONS: Significance of our investigation is development of score that help phisicians to recruit lymphoma patients at risk for development of thromboembolic complications. Also, we can say that our score is dynamic allowing us to change approach during different phase of therapy and is not limited to outpatient settings or with some complicated laboratory analysis.

Orbital and ocular adnexal Mucosa-Associated Lymphoid Tissue (MALT) lymphomas: a single-center 10-year experience.

Orbital and ocular andexal Mucosa-Associated Lymphoid Tissue Lymphoma (MALT) or ocular adnexal MALT lymphoma (OAML) is the most common of all eye non-Hodgkin lymphomas. Autoimmune inflammatory disorders and chronic infections are important etiological factors and CD5 and CD43 (sialophorin) tumor markers are significant negative prognostic factors. Disease signs and symptoms can occur a long time before diagnosis. Varieties of treatment options are available. The aim of this retrospective analysis was to compare the efficiency of different treatment options and to investigate disease outcome. Twenty OAML patients, diagnosed in the Clinic of Hematology, Clinical Centre of Serbia, between 2003 and 2013, were enrolled. In most cases, OAML developed in the eighth decade with greater incidence in the male population. Median age was 67.5 years. The median period between the appearance of local signs and symptoms and diagnosis was 7 months. The dominant sign at presentation was swelling of involved tissue (40%). The most common was orbital involvement (55%). All patients had localized disease. Observed laboratory parameters on presentation showed low disease activity. Sialophorin prognostic significance was not registered. Our patients were initially treated differently but there was no significant difference in progression-free survival (PFS) due to initial treatment option (p = 0.2957). Median PFS was 22 months (3-89), and 5-year PFS was 60%. Median overall survival (OS) was 43 months (1-105) and 5-year OS 95%. Eight patients (40%) relapsed and one patient died due to non-hematological complications. In our experience, most modern induction treatment options appear to result in the same, favorable outcome.

Visual cleanliness scores of cattle at slaughter and microbial loads on the hides and the carcases.

In two abattoirs, visual cleanliness of 100 cattle was assessed before slaughter (on a scale of 1 to 4). From each animal, two sponge swabs (approximately 2000 cm(2) area, each) were taken: (a) from hide, immediately after sticking, and (b) from final carcase, but before chilling. In each swab sample, total viable count (TVC), Enterobacteriaceae count (EC) and the presence of Escherichia coli O157 were determined. The mean TVC/EC status of hides and final carcases differed significantly only between very dirty cattle (category 4) and all other less dirty or clean cattle (categories 1, 2 and 3), but not between the less dirty and clean cattle (between categories 1, 2 and 3). However, no clear relationship between the visual cleanliness of the hide and the occurrence of E coli O157 on hide or dressed carcases was observed. The study indicated the possibility that visual categorisation of cattle into only two main categories - one containing very dirty animals (category 4 in this work, corresponding to categories 4+5 in The UK Food Standards Agency system) and another containing all the other less dirty or clean animals (categories 1+2+3) - could be sufficient in practice.

Treatment of cattle hides with Shellac solution to reduce hide-to-beef microbial transfer.

The hide-to-beef microbial transfer-reducing effects of a novel Shellac treatment of hides (based on "on-hair immobilization" of microorganisms) were evaluated. In the hide-to-meat direct contact laboratory-based experiments, treatment of hides (of varying visual cleanliness) with Shellac produced significant microbial reductions on beef: up to 3.6 log(10) CFU/cm(2) of total viable count of bacteria (TVC), up to 2.5 log(10) CFU/cm(2) of Enterobacteriaceae (EC) and up to 1.7 log(10) CFU/cm(2) of generic Escherichia coli (GEC). In a small commercial abattoir under "bad-case" conditions (slaughtering dirty cattle, inadequate process hygiene), treatment of hides with Shellac produced significant microbial reductions on beef carcasses: 1.7 log(10) CFU/cm(2), 1.4 log(10) CFU/cm(2) and 1.3 log(10) CFU/cm(2) of TVC, EC and GEC, respectively. In both laboratory- and abattoir-based trials, TVC reductions on beef achieved by the Shellac hide treatment were superior to those achieved by the comparative sanitizer rinse-vacuum hide treatment, but reductions of EC and GEC did not differ significantly between the two hide treatments.

Intravascular large B-cell lymphoma of central nervous system - a report of two cases and literature review.

Intravascular large B-cell lymphoma (IVL) is a rare form of diffuse large B cell lymphoma (DBCL) frequently presenting with skin and/or central nervous system (CNS) involvement. IVL involves CNS in 75 - 85% of patients and neurological symptoms include sensory and motor deficits or neuropathies, meningoradiculitis, paresthesia, hypostenia, aphasia, dysarthria, hemiparesis, seizures, transient visual loss, vertigo and impaired cognitive function. Neuroimaging discloses CNS involvement only in half of patients with neurological symptoms because there are no pathognomonic neuroradiological findings for IVL; ischemic foci are the most common presentation pattern and therefore vasculitis is the most common differential diagnosis. According to all mentioned data, diagnosis of CNS IVL requires a histopathological confirmation. Brain biopsy is absolutely indicated in patients with progressive neurological deterioration with unclear abnormalities in cerebral MR imaging. A general policy is that patients with IVL should be considered to have disseminated disease and should be treated with systemic chemotherapy. In younger patients with unfavorable features the high-dose chemotherapy with autologous stem cell transplantation should be used. Nevertheless, the course of IVL is rapidly progressive and ultimately fatal.

Treatment of cattle hides with Shellac-in-ethanol solution to reduce bacterial transferability--a preliminary study.

A solution of natural, food-grade resin (Shellac) in ethanol was evaluated to treat samples of visually clean and dry cattle hides with the aim to reduce bacterial removability from the hides by swabbing. Hide treatment by 23% Shellac-in-ethanol solution reduced sponge-swabbing recoveries of general microflora (TVC) by a factor of 6.6 logs (>1000-fold larger than the 2.9 log reduction observed by ethanol alone), and of generic Escherichia coli and Enterobacteriaceae by factors of at least 2.9 and 4.8 logs, respectively. These reductions were superior to those achieved by a sanitizer rinse-vacuum hide treatment. Significantly greater reductions of TVC recoveries from hides were achieved when using higher Shellac concentrations (23 and 30% rather than 4.8-16.7%) and when Shellac solution temperatures were 20-40 degrees C rather than 50-60 degrees C. Furthermore, the Shellac-based treatment also markedly reduced the E. coli O157 prevalence (3.7-fold reduction) on natural, uninoculated hides, as well as the counts of E. coli O157 on artificially inoculated hides (2.1 log reduction). This preliminary study indicated that a "bacterial on-hide immobilisation" approach to reducing transmission of microorganisms from cattle hide is promising and so will be further explored.

Primary extranodal lymphomas of gastrointestinal localizations: a single institution 5-yr experience.

This study is aimed at comparison of patients with extranodal lymphomas based on pathohistological findings differences (MALT vs non-MALT) as well as regarding gastric and non-gastric localization, and determining the significance of clinical-laboratory parameters with respect to therapeutic response and length of survival. A total of 56 patients with extranodal non-Hodgkin's lymphomas of the gastrointestinal tract were evaluated over a 5-yr period. Regarding the localization of the disease, the stomach was most frequently affected, 39 patients (70%); followed by small and large intestines, 17 patients. As for the pathohistological findings, MALT lymphoma accounted for 70%, DLBCL 25%, while other subtypes accounted for 5%. Patients' distribution was analyzed according to CS based on both Ann Arbor and Lugano systems; however, the difference obtained between the groups was not statistically significant in both staging types of patients. Statistically significant difference in patients' distribution was not found with respect to IPI index, bone marrow infiltration, anemia, hypoalbuminemia, or histological subtype and localization. Difference in survival between patients according to the pathohistological type was not statistically significant also according to the type of the affected gastrointestinal tract organ. Statistical significance of difference according to survival probability was obtained based on age (survival is longer in patients over 55 yr of age); according to CS and according to Ann Arbor and Lugano classifications (the patients with lower CS live significantly longer); according to IPI index (the survival is significantly longer in patients with lower probability: IPI-0,1, and 2), as well as patients free of bone marrow infiltration whose survival is also significantly longer.

ELAV tumor antigen, Hel-N1, increases translation of neurofilament M mRNA and induces formation of neurites in human teratocarcinoma cells.

Human ELAV proteins are implicated in cell growth and differentiation via regulation of mRNA expression in the cytoplasm. In human embryonic teratocarcinoma (hNT2) cells transfected with the human neuronal ELAV-like protein, Hel-N1, neurites formed, yet cells were not terminally differentiated. Cells in which neurite formation was associated with Hel-N1 overexpression, also expressed increased levels of endogenous neurofilament M (NF-M) protein, which distributed along the neurites. However, steady-state levels of NF-M mRNA remained similar whether or not hNT2 cells were transfected with Hel-N1. These findings suggest that turnover of NF-M mRNA was not affected by Hel-N1 expression, despite the fact that Hel-N1 can bind to the 3' UTR of NF-M mRNA and was found directly associated with NF-M mRNA in transfected cells. Analysis of the association of NF-M mRNA with the translational apparatus in Hel-N1 transfectants showed nearly complete recruitment to heavy polysomes, indicating that Hel-N1 caused an increase in translational initiation. Our results suggest that the stability and/or translation of ARE-containing mRNAs can be regulated independently by the ELAV protein, Hel-N1, depending upon sequence elements in the 3' UTRs and upon the inherent turnover rates of the mRNAs that are bound to Hel-N1 in vivo.

Messenger ribonucleoprotein complexes containing human ELAV proteins: interactions with cytoskeleton and translational apparatus.

Mammalian ELAV proteins bind to polyadenylated messenger RNAs and have specificity for AU-rich sequences. Preferred binding sites in vitro include the AUUUA pentamer and related sequences present in the 3' untranslated regions of many growth regulatory mRNAs. Human ELAV (hELAV) proteins have been implicated in post-transcriptional regulation of gene expression by their effects on the stability and translatability of growth regulatory mRNAs. We have examined the intracellular localization of ELAV proteins in neurons and in tumor cells of neuronal origin using indirect immunofluorescence, confocal microscopy and biochemical separation. Mammalian neuronal ELAV proteins are found predominantly in the cytoplasm of cells in mRNP complexes termed alpha complexes which, when associated with polysomes, form large and high density ss complexes, as assayed by glycerol and accudenz gradients, respectively. Puromycin, cytochalasin or EDTA treatments disrupt beta complexes causing the release of alpha complexes, which then appear, by confocal microscopy, as large hELAV mRNP granules associated with microtubules. Association of partially purified hELAV mRNP alpha complexes with microtubules was confirmed by in vitro reconstitution assays. Furthermore, colchicine treatment of cells suggested that association of hELAV mRNP alpha complexes with microtubules is also necessary for the formation of ss complexes. Our data suggest a model in which a subset of mRNAs is associated with microtubules as ELAV mRNP particles (alpha complexes) which, in turn, associate with polysomes to form a translational apparatus (beta complex) that is, through polysomes, associated with the microfilament cytoskeletal network. hELAV proteins in these mRNP granules may affect post-transcriptional regulation of gene expression via the intracellular transport, localization and/or translation of growth regulatory mRNAs.

Maturation of Hantaan virus glycoproteins G1 and G2.

Hantaan virus-infected Vero E6 cell lysates were used for immunoprecipitation with monoclonal antibodies against glycoprotein G1 (MAbG1) or G2 (MAbG2). When cell lysates were prepared with buffer containing nonionic detergent, both G1 and G2 glycoproteins were precipitated with either MAbG1 or MAbG2. In contrast, when cell lysates were prepared with a buffer containing ionic detergents MAbG1 precipitated only glycoprotein G1 and MAbG2 precipitated only glycoprotein G2. Heterodimers and possibly higher oligomeric forms of the glycoproteins were detected on nonreducing SDS-polyacrylamide gels only after chemical cross-linking and immunoprecipitation with either MAbG1 or MAbG2. In order to determine the sites of Hantaan virus glycoproteins maturation and the G1-G2 complex formation, infected cells were treated with inhibitors that prevent specific steps of oligosaccharide processing. Furthermore, glycoproteins G1 and G2 immunoprecipitated from infected cell lysates or from isolated virus particles were tested for sensitivity to endoglycosidase H, endoglycosidase F, and endoglycosidase D. The results of these experiments show that maturation of both G1 and G2 takes place in the endoplasmic reticulum (ER). Furthermore, G1-G2 complex formation occurs in the ER as well, since the two glycoproteins co-precipitated with either MAbG1 or MAbG2 from infected cell lysates treated with brefeldin A and prepared with buffer containing nonionic detergent.

Comparison of the deduced gene products of the L, M and S genome segments of hantaviruses.

The amino acid sequences deduced from all currently available nucleotide sequences of hantaviruses are compared. Comparisons of three large (L), eight medium (M) and five small (S) genome segments are included. A consensus sequence is provided, allowing easy identification of conserved and unique gene regions. The viruses included in this report represent four serologically distinct hantaviruses which are capable of causing severe, moderate, mild or no human disease.

Selenium status of patients with Balkan endemic nephropathy.

The selenium (Se) contents in common cereals in endemic and nonendemic areas in Serbia are very low. Plasma Se levels of both patients and healthy subjects, were also low, reflecting low Se intakes. Patients with Balkan endemic nephropathy (BEN) had significantly lower (p less than 0.05) plasma Se levels than healthy individuals, both from regions close to endemic areas, and from Belgrade. Mean plasma Se of BEN patients was slightly but insignificantly higher in samples taken immediately after dialysis than in those taken before, suggesting that very little of the Se present in plasma is dialyzable. Plasma SeGSH-Px activities before and after hemodialysis in both BEN and Nonendemic chronic renal failure (NCRF) patients were not significantly different, but BEN patients had lower enzyme activities than those with NCRF and healthy controls. In BEN patients, a significant correlation between plasma Se and SeGSH-Px activity was found.

Molecular characterization of the M genomic segment of the Seoul 80-39 virus; nucleotide and amino acid sequence comparisons with other hantaviruses reveal the evolutionary pathway.

The genomic M segment of Seoul 80-39 virus was characterized by cloning and nucleotide sequence analysis. The virion M RNA segment is 3651 nucleotides long with the 3' and 5' terminal sequences inversely complementary for 20 bases. A single open reading frame was detected in the viral complementary-sense RNA which can encode a polypeptide of 1133 amino acids. The Seoul 80-39 virus M segment was compared with the M segments of related viruses, Hantaan 76-118, Hallnas B1 and Sapporo Rat (SR-11) virus. Our results demonstrate a significant similarity between M RNA segments of the Seoul 80-39, Hantaan 76-118, Hallnas B1 and SR-11 viruses. The degree of conservation of both nucleic acid and protein sequences between these viruses reveals a close evolutionary relationship. Furthermore, it is evident that the serotypic profile of hantaviruses is determined by the rodent host species from which the virus was isolated and not by the geographical area.

Nucleotide sequence and coding capacity of the large (L) genomic RNA segment of Seoul 80-39 virus, a member of the hantavirus genus.

The nucleotide sequence of the large (L) genomic RNA segment of Seoul 80-39 virus was determined from overlapping cDNA clones. The virion L RNA segment is 6530 nucleotides long. The 3' and 5' terminal sequences are inversely complementary for 15 bases. The viral complementary-sense RNA contains a single open reading frame from an AUG codon at nucleotide position 37-39 to a UAA stop codon at nucleotide position 6490-6492. This ORF could encode a polypeptide of 2151 amino acids (246,662 kDa) which likely corresponds to the L protein detected in purified viral particles (Elliott et al., 1984) and is assumed to be an RNA-dependent RNA polymerase molecule (Schmaljohn and Dalrymple, 1983). Comparison of the L protein of the Seoul 80-39 virus with the polymerase proteins encoded by other negative-stranded RNA viruses revealed 44% similarity only with the part of the Bunyamwera virus L protein (Elliott, 1989) and a very weak homology with the PB1 protein of influenza virus.

[Value of the fetal biophysical profile in intensive monitoring of high risk pregnancy]. / Pouzdanost biofizickog profila ploda u intenzivnoj kontroli visoko rizicnih trudnoca.

The fetal biophysical profile (FBP) is a combination of both acute and chronic condition markers of the fetus in the uterus (fetal movements, fetal breathing movements, amniotic fluid volume, maturity of the placenta, intestine distension, cardiotocographic "non stress test", cardiotocographic "oxytocin stress test" cardiotocographic "physical stress test") presented numerically, is an irreplaceable method for monitoring the condition of the fetus in the uterus. In 271 women with high risk pregnancies at the gestational age of 34-44 weeks, depending on the FBP value, the time and mode of pregnancy termination were chosen. There is a significant difference regarding the outcome of pregnancy (general bad outcome--p less than 0.001, mortality--p less than 0.004, low 5 min Apgar score--p less than 0.002, fetal distress during labour--p less than 0.001) depending on the FBP value (normal or pathological) and the connection of the pathological FBP values with the low fetal biochemical profile values (estriol in 24th urine less than 40,000 n mol/day and HPL less than 5 mg). Sensitivity (46.4%), specificity (99.2%), the prognostic value of the pathological test PVP (86.7%), and prognostic value of the normal test - PVN (94.1%) show a high FBP value as a diagnostic test. It is necessary to stick to the protocol while using this test which is very simple, cheap and not at all harmful.