Trace metals in the coastal soils developed from estuarine floodplain sediments in the Croatian Mediterranean region.

Fertile soils in the River Neretva estuary were developed by fluvial sedimentation and deposition of the eroded soil material from the karst hills within the catchment. After extensive reclamation, two reclaimed land zones (fluvial terraces and lower-laying terraces) have been delineated, both used for agriculture. The main objectives of this study were to evaluate soil chemical and geochemical properties in reclaimed zones that differ mainly in topography, soil types and agricultural land use. The origin of the trace metals in the arable soils was studied using multivariate statistics, and interpolation maps of trace metals were produced using GIS and geostatistics. Soil trace metal concentrations do not exceed a threshold value established by the Croatian Government regulation, with exception of copper. Comparative analysis of the main soil properties and trace metal concentrations in the study area showed a pronounced spatial variation and differences between two reclaimed zones in soil organic matter content, bioavailable P and total concentrations of Cd and Cu. Factor analysis in the area of the lower-laying terraces showed grouping of bioavailable P and K, organic matter content and pH (negative loading) in the component associated mostly with the land use. In the area of the fluvial terraces, bioavailable P and total Cd were grouped in the same component that may be explained by the traditional small farm agriculture and overuse of mineral fertilizers. In the whole study area, processes of secondary salinization were determined, accompanied by the raised chloride and sodium concentration measured in the saturation soil extract.

[10 years' of production and use of human rabies immunoglobulin in Yugoslavia]. / Deset godina proizvodnje i primene humanog imunoglobulina protiv besnila u Jugoslaviji.

Application of the rabies immunoglobuline is a compulsory part of the prophylaxis of rabies in all severe, transdermal lesions caused by rabies infected animals. Sylvatic rabies has spread in the past few years throughout the whole Yugoslavia, and human cases of rabies have also been reported in other East European countries. In order to achieve the highest level of rabies prophylaxis, apart from postinfective rabies vaccination, it is necessary to provide passive immunization using specific antibodies against rabies. After successful immunization of the young, healthy volunteers in 1990, National Blood Transfusion Institute, in cooperation with the Pasteur Institute from Novi Sad, prepared the first quantities of immunized plasma by plasmapheresis procedure and human rabies immunoglobuline. Without national production, sufficient quantities of human rabies immunoglobuline could not be provided, since the price on the world market is rather high (over $1000 per patient).

A simple PCR with different 3' ends of the third primer for detection of defined point mutations: HCV genotyping as an example.

In this article, we describe a useful modification of the polymerase chain reaction for amplification applicable to hepatitis C virus genotyping and determination of its subtypes. The method is fast, cheap and simple for detection of any known point mutation, and could be used in every laboratory with experience in polymerase chain reaction technique. We could differentiate hepatitis C virus subtype 1b from other subtypes and 2b from 2a and other subtypes as well. We could also differentiate hepatitis C type 3 using a type-specific oligonucleotide from 3a subtype, thus covering the most common hepatitis C virus (sub)types present in the European region.

Hepatitis C virus RNA testing by nested PCR in blood preparations in Yugoslavia.

Patients receiving any kind of human blood preparations are in permanent danger of any infection including hepatitis C (HCV) infection. Testing for the presence of HCV in blood preparations is one of the steps towards safe medical treatment. One of the approaches for this testing is a detection of HCV nucleic acid. In this paper we describe a simple method for isolation of HCV RNA from blood preparations and control of HCV RNA presence in 19 intravenous and intramuscular products, manufactured in the National Blood Transfusion Institute in Belgrade. RT-PCR was performed according the rules saving RNA. Primers were located in 5' conserved region. Seven out of 19 batches of gamma-globulin, albumin, anti-tetanus and anti-rabies immunoglobulin preparations were found to be HCV RNA positive. For the time being, the PCR method is too expensive for routine HCV RNA testing of hundreds of blood donors per day. Serological screening test of blood donors and nested PCR testing for HCV RNA in blood preparations could be an efficient combination of tests in prevention of posttransfusion hepatitis C.

[Detection of the presence of the hepatitis C viral genome in immunoglobulin preparations using RT-PCR]. / Provera prisustva genoma virusa hepatitisa C u imunoglobulinskim preparatima pomocu metode RT-PCR.

Contemporary methods of molecular genetics were used to investigate the presence of hereditary matter, i.e., virus hepatitis C genome in immunoglobulin preparations of the Institute for blood transfusion of the Republic of Serbia. ELISA test in immunoglobulin preparations indicated the presence of antibodies to an antigen of hepatitis C virus. After RNA isolation and reverse transcription (RT), double reaction of in vitro DNA amplification (PCR) was done using two pairs of oligonucleotide. After several repeated tests and positive control from blood of the diseased it was concluded that neither of 11 investigated immunoglobulin preparations contained the nucleic acid (RNA) of the HCV origin, that meant that all preparations could be used with no danger of virus hepatitis C infection. Regarding the current experience in relation to the use of PCR for testing of contamination by hepatitis C virus in preparations from human blood, that are used in the therapy of various conditions and diseases, it is recommendable to use this method due to its sensitivity and specificity.

[Contamination of IgG preparation for intravenous use with IgA. I. Development of ELISA tests for determination of low concentrations of IgAc, IgA1, IgA2, IgG and IgM]. / Kontaminacija imunoglobulinima A preparata imunoglobulina G3A intravensku upotrebu. I) Razvoj postupaka enzimskog imunoeseja za merenje niskih koncentracija imunoglobulina A1, A2, G i M.

Very low amounts of immunoglobulin contaminants present in preparations of immunoglobulin G for intravenous use (IVIG) require sophysticated procedures for detection of such low concentrations in milieu of comparatively very high level of IgG. Standard RID and nephelometry procedures are not adequate for these purposes. In process of removal of IgA contamination from IVIG, which is currently under development in Blood Transfusion Institute, Belgrade, it is necessary to have senzitive and very specific procedures for determination of and follow up of very low concentrations of IgA remauning in IVIG. We have established a highly senzitive and specific ELISA precedures for determination of very low concentrations of total IgA (IgAc), IgA1, IgA2, IgG and IgM. Commercial standards of IgAc, IgA1, IgA2, IgG and IgM were bound to PVC plates coated with corresponding monoclonal antibodies anti IgAc-IgA1, anti-IgG and anti-IgM. Resulting standard curves have shown high correlation coeficients with limited range of detection (two orders of magnitude in nanogram range). Very high specificity of ELISA tests were obtained due to the highly specific monoclonal antibodies used. Very high specificity and very low detection level of developed tests are advantageous compared to standard procedures for Ig concentration determination.

[Contamination of IgG preparation for intravenous use with IgA. II. Removal of IgA1 from IgG preparation using affinity chomatography on lectin jacalin columns]. / Kontaminacija imunoglobulinima A preparata imunoglobulina G3A intravensku upotrebu. II) Uklanjenje imunoglobulina A1 iz preparata imunoglobulina G pomocu afinitetne hromatografije na koloni lektina zakalina.

Administration of preparations of IgG for intravenous use (IVIG) in individuals with high anti-IgA antibody levels in circulation is frequently accompained with anafilactoid reactions. Thus, there is a requirement for preparations of IVIG with very low levels of IgA. Reduction of IgA concentration in IgG preparations is generally difficult to achieve, since physical and chemical characteristics, on which proceses for Ig purification are based on, are similar for these two Igs. In this work the ability of lectin jacalin to selectively bind IgA1 subclass was exploited for removal of IgA1 from IVIG currently under development in Blood Transfusion Institute of Republic Serbia, Belgrade. Concentrations of Igs in this preparation of IVIG were measured by ELISA tests developed for these purposes. The results have shown 280 times decrease of IgA1 content without changes of IgG concentration. Therefore, affinity chomatography on jacalin column is a suitable method for removal of IgA1 subclass from IVIG. Because of a favourable pH and ionic strength conditions used in process, this procedure has no effect on native state of IgG.

[Preparation of human albumin preparations]. / Pripremanje humanih albuminskih preparata.

Structure, physical and biochemical characteristics of albumin have already been investigated and described in full detail. Preparation of albumin solution preparations of various concentrations for human application started more than 40 years ago. Person deserving most credit for it is E. Cohn, who first used ethyl alcohol on low temperatures for the separation of certain fractions among which albumin took the largest part. Many modifications of Cohn's method are used today, though the most frequently used one is modification by Kistler and Nitschman. Some new methods appeared recently, ensuring higher degree of economy and efficacy, better yield of albumin and higher purity of final preparations. One of those methods--chromatography method--is recommended by Swedish authors (J. Curling et al.). Thanks to its multiple physiological functions and stability, albumin takes very important place in therapy of many diseases, both in peace and in states of emergency.