Screening of Native Trichoderma Species for Nickel and Copper Bioremediation Potential Determined by FTIR and XRF.

Soil pollution with heavy metals is a serious threat to the environment. However, soils polluted with heavy metals are considered good sources of native metal-resistant Trichoderma strains. Trichoderma spp. are free-living fungi commonly isolated from different ecosystems, establishing endophytic associations with plants. They have important ecological and biotechnological roles due to their production of a wide range of secondary metabolites, thus regulating plant growth and development or inducing resistance to plant pathogens. In this work we used indigenous Trichoderma strains that were previously isolated from different soil types to determine their tolerance to increased copper and nickel concentrations as well as mechanisms of metal removal. The concentrations of bioavailable metal concentrations were determined after extraction with diethylene-triamine pentaacetate (DTPA)-extractable metals (Cd, Cr, Co, Cu, Pb, Mn, Ni, and Zn) from the soil samples by inductively coupled plasma-optical emission spectrometry (ICP-OES). Two indigenous T. harzianum strains were selected for copper tolerance, and three indigenous T. longibrachiatum strains were selected for nickel tolerance tests. Strains were isolated from the soils with the highest and among the lowest DTPA-extractable metal concentrations to determine whether the adaptation to different concentrations of metals affects the mechanisms of remediation. Mechanisms of metal removal were determined using Fourier-transform infrared spectroscopy (FTIR) and X-ray fluorescence spectroscopy (XRF), non-destructive methods characterized by high measurement speed with little or no need for sample preparation and very low costs. Increased DTPA-extractable metal content for nickel and copper was detected in the soil samples above the target value (TV), and for nickel above the soil remediation intervention values (SRIVs), for total metal concentrations which were previously determined. The SRIV is a threshold of metal concentrations indicating a serious soil contamination, thus confirming the need for soil remediation. The use of FTIR and XRF methods revealed that the presence of both biosorption and accumulation of metals in the Trichoderma cells, providing good bioremediation potential for Ni and Cu.

[Incidence, risk factors and outcome of nosocomial pneumonia patients with central nervous system infections].

INTRODUCTION: Pneumonia is the most frequent nosocomial infection in intensive care units. The reported frequency varies with definition, the type of hospital or intensive care units and the population of patients. The incidence ranges from 6.8-27%. OBJECTIVE: The objective of this study was to determine the frequency, risk factors and mortality of nosocomial pneumonia in intensive care patients. METHODS: We analyzed retrospectively and prospectively the collected data of 180 patients with central nervous system infections who needed to stay in the intensive care unit for more than 48 hours. This study was conducted from 2003 to 2009 at the Clinical Centre of Kragujevac. RESULTS: During the study period, 54 (30%) patients developed nosocomial pneumonia. The time to develop pneumonia was 10 +/-6 days. We found that the following risk factors for the development of nosocomial pneumonia were statistically significant: age, Glasgow Coma Scale (GCS) score < 9, mechanical ventilation, duration of mechanical ventilation, tracheostomy, presence of nasogastric tube and enteral feeding. The most commonly isolated pathogens were Klebsiella-Enterobacter spp. (33.3%), Pseudomonas aeruginosa (24.1%), Acinetobacter spp. (16.6%) and Staphylococcus aureus (25.9%). CONCLUSION: Nosocomial pneumonia is the major cause of morbidity and mortality of patients with central nervous system infections. Patients on mechanical ventilation are particularly at a high risk. The mortality rate of patients with nosocomial pneumonia was 54.4% and it was five times higher than in patients without pneumonia.

Characteristics of radiological changes in lungs during varicella zoster viral infection.

The objective of this research was to analyse the varicella patients' data in order to determine the following: general frequency of pneumopathies and types of lung changes according to X-ray presentation and the changes on the computed tomography. It examined 101 patients with the clinical presentation of varicella and some of the X-ray entities of varicella pneumopathies. Radiological techniques included chest X-ray and CT scans. Familiarity with clinical, laboratory and radiological characteristics of the disease may be of utmost importance for early recognition.

[Parameters for assessing the severity of clinical presentation of hemorrhagic fever with kidney syndrome].

INTRODUCTION: Hemorrhagic fever with kidney syndrome is an acute infectious disease of the zoonotic character, which is characterized by the sudden beginning, high temperature, distinct hemorrhagic syndrome and symptoms and signs of the acute kidney insufficiency. Hemorrhagic fever with kidney syndrome can have mild, moderate and severe clinical picture and the mortality ranges from 5% -15% in our area. The purpose of this study was to try to present criteria on the basis of which it would be possible to forsee, on time, the severity of the clinical picture and to recognize the patient with the most severe disease form. MATERIAL AND METHODS: Within the period often years, 87 patients became ill of hemorrhagic fever with kidney syndrome and were treated at General Hospital in Berane, where they were followed from the aspect of clinical and laboratory analyses while the serologic confirmation of the disease was done at the Institute of Immunology and Virology "Torlak" in Belgrade. RESULTS: The results show that the severity of the clinical picture can be forseen on time on the basis of the dynamics of increase of the values of urea and creatinine, duration of the shock condition, duration of the oliguria less than 300 ml/24 h, number of the collapses, presence of the meningitic syndromes and appearance of the asciteses and pleuritic hemorrhage. When, on the basis of these parameters, we classify the patients into the categories of mild, moderate and severe clinical picture, using the appropriate statistics methods we find out the differences in duration of the disease as well as in the result of treatment and number of the dialysed patients. CONCLUSION: It is possible to forsee clinical course in a patient on time by making timely diagnosis and careful following the clinical and laboratory parameters, important for the result of treatment.

[Lyme borreliosis--diagnostic difficulties in interpreting serological results].

INTRODUCTION: Lyme borreliosis is a multi-systemic disease caused by spirochete Borrelia burgdorferi sensu lato. The specific response is influenced by phenotypic characteristics of Borrelia, different antigen structure, their different geographic distribution, and the patient's capability to react to the infection. The immune response to Borrelia burgdorferi sensu lato develops relatively late, whereas in some patients it never develops. The immune response in the early phase of Lyme borreliosis is very similar to the one of healthy population. DIAGNOSIS OF BORRELIA: Clinical manifestation, detailed anamnesis and epidemiological data are crucial for making the diagnosis. The majority of patients in the late phase of Lyme borreliosis have IgG antibody response, which could be followed by IgM also throughout this period of time. The number of serologically positive findings increases with the duration of the infection. Specific borrelial antigens can be detected by a Western blot test. In patients with neuroborreliosis, antibodies could be synthesized only intrathecally. IgG and IgM antibody response can persist for many years after the treatment. There is no positive serological test, which could be the indicator of the disease activity on its own; even if it demonstrates high antibody titre. If there are no clinical signs of Lyme borreliosis, the diagnosis of Lyme borreliosis should be primarily based on clinical findings, and serological results should be used only to confirm but not to make the diagnosis of Lyme borreliosis. Specific antibodies from the IgM class can be proved in about 50% of patients, 2 to 4 weeks after the onset of primary infection, but an early administration of the antibiotics can postpone or inhibit that response. INTERPRETATION OF SEROLOGICAL RESULTS: When interpreting the serological test results with high level of sensitivity and specificity used for making diagnosis of Lyme borreliosis, it is necessary to take into consideration the seroprevalence in a certain region. In the population with a low prevalence of the disease, the tests will have a low positive predicative value, i.e. the probability of indicating the real disease will be lower. According to the recommendations given by the Centre for Disease Control in North America, all extreme and positive results of EA and IFA are to be confirmed by a Western blot test. DIAGNOSTIC PROBLEMS: The main problem in making diagnosis of Lyme borreliosis is underestimation and overrating of the diagnosis. Not a single positive serologic test is the indicator of the disease activity on its own, regardless of the antibodies titre level, when clear clinical signs are scarce.

[Risk factors and disposition in development of the nervous system infections].

INTRODUCTION: Although well protected, brain is not resistant to infection agents. Acute infections of our nervous system appear more often in children and in persons who have medical history data about previous disorders, especially disorders of the nervous system. It is difficult to list possible risk factors which can be responsible for the appearance of infections of CNS and the resulting conditions. It is often difficult or impossible to determine what previous neural damage was (trauma, anoxic damages etc.) from those appearing during infections of CNS. All-inclusive anamnestic research reduces the possibility of approximate judgements. MATERIAL AND METHODS: The research was based on the retrospective analysis of medical documentation of 275 patients. All patients were divided into three groups according to the final diagnosis. The first group consisted of 125 patients who were treated for acute virus encephalitis, the second group consisted of 125 patients who were treated for acute bacterial meningoencephalitis and the third group consisted of 25 patients who were treated for cerebritis. DISCUSSION: In our studies sample, the youngest patient was 3 years old and the oldest was 87 years old. The highest number of patients with virus infection of the CNS was in the group under 25 years of age (45.6%). The highest number of patients with bacterial infections of the CNS and cerebritis was in the group of patients over 45 years of age (64%, 37%). CONCLUSION: Risk factors were more present in bacterial infections of the nervous system and cerebrit than in virus infection of CNS. In virus infections of the CNS, 28% of patients had some risk factor, most often-chronic ethylism, diabetes mellitus and acquired heart diseases. In bacterial infections of the CNS, 64% of patients had some predisposed factor. The most frequent factor of risk in these patients were chronic otitis (21.6%) and cranio-trauma (14.4%). In cerebritis, risk factors were present in 76% of patients and they were: sepsis (20%), chronic otitis (12%) and systemic lupus erythematosus (8%).

[Altered state of consciousness as a factor affecting the course and consequences of acute viral encephalitis].

INTRODUCTION: Acute viral encephalitis is an infectious disease, which is characterized by sudden onset, serious clinical picture and with an uncertain outcome. Disturbances of consciousness are common in the course of acute viral encephalitis. Consciousness is the heighest form of psychic life and consciousness disturbances may include quantitative (somnolentia, sopor, coma) and qualitative symptoms (convulsions, twilight state). MATERIAL AND METHODS: We investigated 63 patients with acute viral encephalitis. The youngest patient was 15, and the oldest one 69 years of age. We examined the state of consciousness in all patients. RESULTS: Of all examined patients, 54 (85.72%) patients had disturbances of consciousness, 9 (14.25%) patients had no conciousness disturbances, 11 (17.46%) patients had sopor and 24 (38.09%) patients were in coma. The highest percentage of lethal outcomes was recorded in coma patients (12.5%). The longer the duration of consciousness disturbances, the higher the mortality among these patients. DISCUSSION: Subjective and objective factors cause different grades and duration of consciousness disturbances. The highest percentage of patients who recovered without consequences was established in the group of patients (81.82%) with somnolence. The highest percentage of patients who recovered with consequences (20.8%) and patients (12.5%) with lethal outcome was recorded in the group of patients in coma. These findings are in correlation with literature data.

[Changes in plasma osmolality in food poisoning].

INTRODUCTION: Changes in plasma osmolality may occur during acute intestinal infections due to dehydration (loss of water and/or electrolytes). Depending on whether the water and electrolyte deficit is primary, or a proportional loss of water and electrolytes, dehydration can be classified into three categories: hypertonic, hypotonic and isotonic. MATERIAL AND METHODS: Thirty (30) patients with food poisoning were included in this research. All patients were hospitalized because of frequent vomiting, with resultant dehydration. A diagnosis of food poisoning was made based on the clinical picture, short incubation period and positive epidemiological history. Plasma osmolality was measured by a freezing point depression with an osmometer, while effective plasma osmolality was determined by using the following formula: EPO (eff. plasma osmolality) = 2 x serum sodium concentration + serum glucose level. Apart from plasma osmolality, other parameters were also measured in patients' sera: sodium, chloride, potassium, urea, glucose and hematocrit. In order to follow-up the changes in these parameters, they were also measured after treatment of the gastrointestinal disorder. Statistical analysis was performed using the equality of mean values for 2 basic groups. RESULTS: The statistical results showed that the values of total and effective plasma osmolality (TPO and EPO) among patients with gastrointestinal disorders were not significantly higher than values after the alimentary infection. DISCUSSION: Such results suggest that food poisoning is associated with disorders of water and electrolyte metabolism, that is isotonic type of dehydration.

Explorative data analysis of two-dimensional electrophoresis gels.

Methods for classification of two-dimensional (2-DE) electrophoresis gels based on multivariate data analysis are demonstrated. Two-dimensional gels of ten wheat varieties are analyzed and it is demonstrated how to classify the wheat varieties in two qualities and a method for initial screening of gels is presented. First, an approach is demonstrated in which no prior knowledge of the separated proteins is used. Alignment of the gels followed by a simple transformation of data makes it possible to analyze the gels in an automated explorative manner by principal component analysis, to determine if the gels should be further analyzed. A more detailed approach is done by analyzing spot volume lists by principal components analysis and partial least square regression. The use of spot volume data offers a mean to investigate the spot pattern and link the classified protein patterns to distinct spots on the gels for further investigation. The explorative approach in analysis of 2-D gels makes it possible, in a fast and convenient way, to screen many gels in order to determine the protein patterns that form clusters and could be selected for further examination.

[Pseudomembranous colitis during antibiotic therapy]. / Pseudomembranozni kolitis u toku antibiotske terapije.

INTRODUCTION: The use of antibiotics is commonly accompanied by diarrhea: idiopathic diarrhea with a benign process and diarrhea caused by Clostridium difficile and pseudomembranous colitis. Clostridium difficile colonizes the gastrointestinal tract and produces a toxin in cases when normal flora is suppressed by antibiotics. Pseudomembranous colitis most frequently appears after application of clindamycin, lincomycin, ampicillin, cephalosporins and other antibiotics. Diagnosis is established after rectoscopic findings of adherent pseudomembrane and pathohistological verification. The diagnosis is confirmed if there is evidence of Cl. difficile toxin in feces. CASE REPORT: We report about the clinical course of two patients with antibiotic-associated colitis. The diagnosis were made by clinical examinations, rectoscopy and pathohistologic verification of biopsy specimen of the intestinal mucosa. Neutralization test was not done due to technical reasons. Patients were treated with metronidazole. Unwanted side-effects of metronidazole therapy were not observed. DISCUSSION: Both our patients confirmed that they previously used different antibiotics. In the first case, diarrhea appeared during the antibiotic therapy, and in the second case, after finishing it. After antibiotic use, diarrhea appears in 5.30% cases, but fortunately pseudomembranous colitis is rare. However, taking into consideration that pseudomembranous colitis has a severe course and requires urgent treatment, one has to consider the possibility of pseudomembranous colitis when diarrhea appears during and after antibiotic use in order to initiate adequate therapy.

[Risk factors for hepatitis C virus infection]. / Faktori rizika za nastanak hepatitis C virusne infekcije.

INTRODUCTION: Hepatitis C viral infection represents a major health problem in the world. The estimated global incidence is about 3%, whereas the number of chronic hepatitis C virus (HCV) carriers worldwide is estimated to be between 150-300 million people. MATERIAL AND METHODS: This retrospective analysis included 82 patients whose diagnosis of viral hepatitis C infection was based upon the following criteria: case history, physical examination, laboratory and abdominal ultrasound examination, histological examination of the liver, radiological examination, serological analysis and viral analysis. Descriptive statistics were used to describe general data on patients of the study group, risk factor analysis and follow-up results. RESULTS: The most prominent risk factor in our study group was intravenous use of drugs in 37 patients (37%), and blood transfusion in 13 patients (13%). Less important risk factors of viral hepatitis C infection included: promiscuity (8%), sexual contact with hepatitis C carriers (5%), surgical intervention (5%), hemodialysis (3%), intranasal use of cocaine (2%). DISCUSSION: Hepatitis C viral infection has become the illness of young and middle-aged population. This is due to the epidemic profile of this illness, due to intravenous use of drugs as the most prominent risk factor. CONCLUSION: Due to the number of infected, numerous risk factors and complications of viral hepatitis C, hepatitis C virus has become the most prominent hepatotrophic virus.

Comparison of ethanol-soluble proteins from different rye (Secale cereale) varieties by two-dimensional electrophoresis.

The major storage proteins from six rye varieties, grown under the same conditions in 1997 and 1998 in Rønhave, Denmark, were analyzed by two-dimensional (2-D) polyacrylamide gel electrophoresis. The proteins were extracted from ground rye kernels with 70% ethanol and separated by 2-D electrophoresis. The gels were scanned, compared using ImageMaster software and the data sets were analyzed by principal component analysis (PCA) using THE UNSCRAMBLER software. Afterwards MATLAB was used to make a cluster analysis of the varieties based on PCA. The analysis of the gels showed, that the protein patterns (number of different proteins and their isoelectric points and molecular weights) from the six rye varieties were different. Based on the presence of unique cultivar-specific spots it was possible to differentiate between all six varieties if the two harvest years were investigated separately. When the results were combined from the two years five varieties could be differentiated. The results from the PCA confirmed the finding of the unique spots and cluster analysis was made in order to illustrate the results. The combination of the results from 2-D electrophoresis and other grain characteristics showed that one protein spot was located close to the parameters bread volume and bread height.

Determination of wheat quality by mass spectrometry and multivariate data analysis.

Multivariate analysis has been applied as support to proteome analysis in order to implement an easier and faster way of data handling based on separation by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. The characterisation phase in proteome analysis by means of simple visual inspection is a demanding process and also insecure because subjectivity is the controlling element. Multivariate analysis offers, to a considerable extent, objectivity and must therefore be regarded as a neutral way to evaluate results obtained by proteome analysis. Proteome analysis of storage proteins from the wheat gluten complex based on two-dimensional electrophoresis and analysis of the N-terminal sequence has revealed a protein homologous to gamma-gliadins, tentatively associated with quality and within the molecular weight range 27-35 kDa. Further examinations of gliadin data based on mass spectrometry revealed that quality among wheat varieties could be determined by means of principal component analysis. Further examinations by interval partial least squares made it possible to encircle an overall optimal molecular weight interval from 31.5 to 33.7 kDa. The use of multivariate analysis on data from mass spectrometry has thus shown to be a promising technique to minimize the number of two-dimensional gels within the field of proteome analysis.