Combined early treatment with chelating agents DMSA and CaDTPA in acute oral cadmium exposure

The influence of chelating agents: meso-2,3-dimercaptosuccinic acid (DMSA); calcium trisodium diethylenetriaminepentaacetate (CaDTPA) and their combination on mobilisation of cadmium (Cd) was compared in female albino rats. After oral Cd administration chelators were applied either orally (DMSA) or intraperitoneally (CaDTPA) at various short time intervals after Cd. Three experiments were carried out with four treatment groups in each: 1) Cd (control); 2) Cd+DMSA; 3) Cd+CaDTPA; 4) Cd+DMSA+CaDTPA. Time intervals for chelator treatment after Cd administration were: immediate application in the first, half an hour in the second and one hour in the third experiment. At the end of each experiment cadmium was analysed in kidney and liver. Additionally in experiment 3 essential elements (Fe, Cu, Zn) were also determined in the same organs. In experiment 2 the effect of the treatment on urinary elimination of cadmium, copper and zinc were analysed. Results showed that the efficiency of Cd removal from the body (kidneys and liver) is lower when the time between Cd and chelating agents administration is longer. The two chelators differ in efficiency in mobilizing Cd, with DMSA being more efficient than CaDTPA. The combined therapy with the two chelators gave generally better results. It seems that DMSA which is given orally after oral Cd administration removes this element very efficiently from the gastrointestinal tract. CaDTPA, however, which is given parenterally removes absorbed Cd less efficiently, Organs are not significantly depleted in iron and copper after chelation treatment. Only zinc concentration was, however, significantly lower in the liver and higher in kidneys only after CaDTPA and combined DMSA+CaDTPA chelation.     

Rapid prenatal diagnosis of sickle cell diseases using oligonucleotide ligation assay coupled with laser-induced capillary fluorescence detection

Prenatal diagnosis of sickle cell diseases has been available for several years, and our laboratory has performed over 1000 prenatal diagnoses. However, currently available techniques are labor-intensive and time-consuming, and thus the diagnosis is delayed, making the mother's decision difficult. We describe a rapid, high-throughput technique based on the ligation assay coupled with automated capillary fluorescence detection. This new approach allows the diagnosis of both Hgb S and Hgb C to be available in a few hours. We have utilized this technique in 30 prenatal diagnoses and found it to be in complete agreement with the standard diagnoses.     

Schedule-dependent interaction between vinblastine and cisplatin in Ehrlich ascites tumors in mice

Information on the in vivo antitumor efficiency of the combination of Vinca alkaloids in animal tumor models, especially vinblastine (VLB) with cisplatin [cis-diamminedichloroplatinum(II); CDDP] is very limited. Therefore, the aim of our study was to explore whether antitumor schedule dependence exists for the combination of CDDP and VLB on i.p. Ehrlich ascites tumors in mice. Animals were treated 3 days after tumor transplantation with VLB (0.006 mg/kg) or CDDP (0.05 mg/kg) alone, VLB followed by CDDP, and CDDP followed by VLB. The time interval between i.p. injections of the drugs was 24 h. Cell number was measured by counting viable cells using the trypan blue exclusion assay, cell platinum content by electrothermal atomic absorption spectrometry, DNA distribution pattern using flow cytometry, apoptosis by flow-cytometric terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, and cell morphology. Combination of CDDP and VLB resulted in additive interaction when VLB preceded CDDP as determined from cell survival data 24 h after completion of the therapy and in increased platinum content (two times) compared with the same combination in a reverse schedule (CDDP given before VLB), which resulted in antagonism. None of the treatment combinations induced apoptosis. We propose that the observed increase in antitumor effectiveness is mainly due to higher platinum accumulation in tumor cells, which we unambiguously demonstrated by measurement of platinum content in the tumor cells, leading to increased cytotoxicity as well as to cell cycle-dependent effects of VLB and CDDP.     

Possibility of application of new pneumococcal conjugate vaccines in children in Slovenia

The emergence of pneumococcal strains resistant to penicillin caused a lot of problems in the therapy of invasive diseases, and added new dimensions to the role of immunisation. In addition to the currently available 23-valent pneumococcal polysaccharide vaccine (PPV) and a new 7-valent conjugate vaccine (PCV) (Prevnar, Wyeth Lederle), two new conjugate vaccines—a 9- and a 11-valent—are being developed. So far, the choice of most appropriate vaccines has depended on the established prevalence of serotypes causing invasive diseases and their antibiotic resistance in the Slovene children population. Between 1993 and 2001, 263 invasive pneumococcal strains isolated from children with invasive diseases were typed. During the period 1998–2001, the same 161 invasive strains were tested for their antibiotic sensitivity. Streptococcus pneumoniae was identified as the major cause of invasive bacterial diseases in the Slovene children population, especially in children under 4 years of age. Distribution by age groups showed the highest incidence in children aged 0–1 years. The predominant serotypes in all age groups were serotypes 14, 1, 19F, 23F, 6B, 18C and 6A. The distribution of penicillin-intermediate and penicillin-resistant strains showed the predominance of serotypes 23F, 14 and 19F. As concerns infection with S. pneumoniae serotypes, we have proved that children aged less than 5 years are more likely to be infected with penicillin-nonsusceptible or intermediate susceptible strains than older children. The 7-valent conjugate vaccine covers 74% of invasive strains in toddlers, but is less effective in older children.

We can conclude that the 9-valent vaccine formulation is optimal for our country, but further cost-effectiveness analysis must be done for recommendation of wide use.

At that moment it is reasonable to use the 7-valent conjugate vaccine for children with chronic cardiovascular, pulmonary, urinary and liver diseases, with asplenia, neoplasmia, diabetes, meningomyelocoele, before or after bone marrow transplantation and in cases of immunodeficiency.     

Level of Knowledge about Sexuality of People with Mental Disabilities

The paper examines the level of knowledge about sexuality of people with mental disabilities. The research also presents differences resulting from sex and the level of mental retardation. The sample included 24 persons with mental disabilities, who the author of this paper knew very well due to the time spent with them during a summer camp at a club in Vjeverica. Data was collected through direct interviews. Two types of questionnaires on sexuality of people with mental disabilities were used during these interviews. Results indicate that the level of knowledge about sexuality is not sufficient. Specifically low knowledge was shown in the area of the protection of sexual health such as sexually transmitted diseases and methods of protection. A relatively good level of knowledge was shown by the respondents in distinguishing between appropriate and inappropriate ways of sexual behaviour and social understanding of certain situational norms. Differences regarding sex and the level of disability were found. The results indicate the need for additional education on sexuality of both people with mental disabilities and their parents, along with support. Despite the fact that the research was conducted on a small occasional sample, it indicates that further research on this subject is needed.     

The prevalence of oral mucosal lesions in a population in Ljubljana, Slovenia

The purpose of the present study was to evaluate the prevalence of oral mucosal lesions and conditions in a population in Ljubljana, capital of Slovenia. A total 1609 subjects represented the study population in the survey about the periodontal treatment needs in a population in Ljubljana, conducted from 1983 to 1987. Ten years later the same 1609 subjects were invited to the second examination. Altogether, 555 (34.5%) of the invited subjects in the age range 25-75 years came for an interview and clinical examination at the Department of Oral Medicine and Periodontology of the Dental Clinic in Ljubljana. Oral mucosal lesions and conditions were evaluated according to the WHO Guide to Epidemiology and Diagnosis of Oral Mucosal Diseases and Conditions. The results showed the presence of one or more mucosal lesions in 61.6% of the population. Fordyce's condition was observed the most frequently (49.7%) followed by: fissured tongue (21.1%), varices (16.2%), history of herpes labialis (16.0%), history of recurrent aphthae (9.7%), denture stomatitis (4.3%), leukoplakia (3.1%), cheek biting (2.7%), lichen planus (2.3%), frictional keratosis (2.2%), geographic tongue (2.2%), geographic and fissured tongue together (1.1%), mucocele (0.9%), smoker's palate (0.5%) and angular chelitis (0.4%). In the population examined, no oral malignancies were observed. Mucosal lesions like whitish lesions, denture related lesions, fissured tongue, varices and mucocele were more prevalent with increasing age. Tobacco-related lesions (leukoplakia and smoker's palate together) were significantly more prevalent among men than among women (P<0.05), while lichen planus, denture stomatitis and herpes labialis occurred more frequently in the female population.     

Low-oxygen and high-carbon-dioxide atmosphere improves the conservation of hematopoietic progenitors in hypothermia

BACKGROUND: During short-term storage of hematopoietic cells (HCs) at 4°C a substantial decline in number and in functional capacity of progenitors occurs after 3 days. We hypothesized that physiologic O₂ and CO₂ concentrations of hematopoietic tissue microenvironment (approx. 3% O₂ and approx. 6% CO₂) could improve cell viability and functionality during storage at 4°C.
STUDY DESIGN AND METHODS: Mobilized peripheral blood (PB) CD34+ cells from multiple myeloma or non-Hodgkin's lymphoma patients were stored in flasks containing air (approx. 20% O₂ and approx. 0.05% CO₂) or 3% O₂/6% CO₂ atmosphere, for 3, 5, and 7 days at 4°C. The total number of cells, the number of cells in G0 or G1 phase of cell cycle, and the apoptosis rate were determined. The functional capacity of stored cells was assessed by the capacity of progenitors to form colonies in methylcellulose (colony-forming cells [CFCs]) and of stem cells to repopulate the bone marrow (BM) of immunodeficient mice (SCID-repopulating cell [SRC] assay).
RESULTS: The total number of viable cells and cells in G1 phase as well as the number of total CFCs were significantly higher at 3% O₂/6% CO₂ than in air at all time points. Cells in G0 phase and SRC were equally preserved in both conditions.
CONCLUSION: Atmosphere with low O₂ and high CO₂ concentration (3% O₂/6% CO₂) in hypothermia (+4°C) during 7 days of storage prevents cell damage and preserves a high number of functional HSCs and progenitors mobilized in PB by granulocyte–colony-stimulating factor.     

Induced synthesis of albumin-like protein in damaged rat reticulocytes

In this study, we present evidence that red blood cell (RBC) membrane p68 in the Belgrade (b/b) rat is similar if not identical to rat serum albumin (RSA). Structural homology between RSA and the RBC p68 has been determined by a variety of biochemical and immunological criteria. This albumin-like protein is a normal constituent of rat RBC and it is partially exported by exosomes during erythroid differentiation. The endogenous origin of rat RBC albumin-like protein was demonstrated by monitoring protein synthesis in reticulocytes and by identification of reticulocyte mRNA for albumin. Haemolytic anaemia, either hereditary (b/b rat) or drug-induced (phenylhydrazine-treated normal rat), results in increased accumulation of the albumin-like protein in rat RBCs as a result of its induced synthesis.     

Molecular mechanisms of insulin resistance and associated diseases

Insulin resistance is a state in which higher than normal concentrations of insulin are required for normal response. The most common underlying cause is central obesity, although primary insulin resistance in normal-weight individuals is also possible. Excess abdominal adipose tissue has been shown to release increased amounts of free fatty acids which directly affect insulin signalling, diminish glucose uptake in muscle, drive exaggerated triglyceride synthesis and induce gluconeogenesis in the liver. Other factors presumed to play the role in insulin resistance are tumour necrosis factor alpha, adiponectin, leptin, IL-6 and some other adipokines. Hyperinsulinaemia which accompanies insulin resistance may be implicated in the development of many pathological states, such as hypertension and hyperandrogenaemia. Insulin resistance underlies metabolic syndrome and is further associated with polycystic ovary syndrome and lipodystrophies. When beta-cells fail to secrete the excess insulin needed, diabetes mellitus type 2 emerges, which is, besides coronary heart disease, the main complication of insulin resistance and associated diseases.     

Using Bayesian methods to test mediators of intervention outcomes in single-case experimental designs

Abstract

Single-Case Experimental Designs (SCEDs) have lately been recognized as a valuable alternative to large-group studies. SCEDs form a great tool for the evaluation of treatment effectiveness in heterogeneous and low-incidence conditions, which are common in the field of communication disorders. Mediation analysis is indispensable in treatment research because it informs researchers about the mechanism through which the intervention leads to changes (e.g., communication skills) in the outcome of interest (e.g., developmental outcomes). Despite the increasing popularity of both SCEDs and mediation analysis, there are currently no methods for estimating mediated effects for a single individual. This paper describes how Bayesian piecewise regression analysis can be used for mediation analysis in SCEDs. A Playskin Lift^TM dataset from one infant born preterm who is at risk for cognitive developmental delays is used to illustrate two approaches to mediation analysis in SCEDs: Bayesian computation of the mediated effect and Bayesian informative hypothesis testing. Annotated R code is provided so researchers can easily fit the proposed models to their own SCED data set. Advantages and limitations of the method are discussed.