The Application of Sequential Extraction in Phosphorus Fractionation in Environmental Samples.

BACKGROUND: Maintaining the balance of phosphorus in ecosystems is a priority activity in environmental protection. To achieve this successfully, it is necessary to know the physicochemical properties of phosphorus forms. OBJECTIVE: Speciation analysis is used for this purpose. This process helps to assess the concentration of various physical and chemical forms of phosphorus. When there are very unstable chemical forms of the analyte hindering the determination in the environmental test, fractionation is performed, i.e., the process of classifying the analyte or a group of analytes from a given sample according to physical properties. METHODS AND RESULTS: The paper discusses the phosphorus fractionation methods in environmental samples. The authors critically analyzed over a dozen phosphorus fractionation procedures using sequential extraction of this element. Particular attention was paid to the possibility of sequential extraction when determining the phosphorus pool available to plants (bioavailable fraction) and labile phosphorus able to migrate in the environment. The paper discusses examples of phosphorus fractionation in various environmental matrices, such as river and lake sediments, sewage sludge, composts, and soils. The selection of an appropriate method for the studied environment was made. CONCLUSIONS: Adapting the proposed methodology to the environmental matrix being tested determines the most accurate results of the experiment. Attention was also paid to the possibilities of practical applications of fractionation results, particularly the possibility of isolating phosphorus available for plants from various environmental matrices.

Resistant Streptococcus pneumoniae strains in children with acute otitis media- high risk of persistent colonization after treatment.

BACKGROUND: Despite advances in the development of pneumococcal conjugate vaccines, acute otitis media (AOM) is a common childhood infection, caused mainly by Streptococcus pneumoniae. It has been suggested that persistence of pneumococcal nasopharyngeal carriage is a risk factor for subsequent recurrent infections. METHODS: In this study we evaluate the relationship between 55 pneumococcal strains obtained from nasopharynx/oropharynx (NP/OP) and middle ear fluid (MEF) of 62 children, aged between 1 and 16 years, during AOM (including recurrent/treatment failure AOM, and post-treatment visits), based on their phenotypic and genotypic characteristics performed by analyses of serotype, antibiotic susceptibility patterns and multilocus sequence typing. RESULTS: S.pneumoniae was isolated from 27.4% of MEF samples; it constituted 43.6% of all positive bacterial samples from MEF samples. There was statistically significant concordance between isolation from the MEF sample and NP/OP colonization by S. pneumoniae (p < 0.0001). During post-treatment visits S.pneumoniae was isolated from 20.8% of children; 91% of them were positive in pneumococcal NP/OP culture during AOM. The serotypes belonging to 10- and 13-valent pneumococcal conjugated vaccines constituted 84% and 92% of the strains, respectively. Multidrug resistance was found in 84% of the strains. According to multivariate analysis, pneumococcal colonization after antibiotic therapy was significantly associated with shorter length of therapy in children with bilateral AOM. CONCLUSIONS: High persistent prevalence of antibiotic-resistant S.pneumoniae strains in children with AOM after unsuccessful bacterial eradication may presumably be regarded as a predisposing factor of infection recurrence.

First report of a Staphylococcus caprae isolated from middle ear fluid of an infant with recurrent acute otitis media.

Staphylococcus caprae was originally isolated from goat milk. This uncommon coagulase-negative staphylococcus, usually associated with animals, has only infrequently been detected in human clinical specimens. Its association with acute otitis media has not been demonstrated so far. The study reports the first isolation of S. caprae from the middle ear fluid of a 12-month-old infant with recurrent, bilateral acute otitis media. Biochemical traits and susceptibility pattern of the isolated strain are also presented.

The role of probiotics in prevention of oral diseases.

The dynamic development of knowledge in the field of probiotics was commenced at the beginning of the 20th century. Since then, many ways of their possible usage in medicine have been established. In accordance with the WHO, probiotics are live microorganisms, which if applied in adequate amounts may benefit the host. Among probiotics, fungi and bacteria are distinguished, and mechanisms of action of these organisms in the oral cavity and gut are parallel. Application in dentistry, in prophylaxis and treatment of oral diseases is still not well known. Most commonly, Lactobacillus reuteri, Lactobacillus rhamnosus GG and Bifidobacterium are applied. The aim of the study was to collect and systematize the latest information about probiotics and their role in pathomechanisms of dental caries, gingivitis and periodontitis, candidiasis, and malodour. Based on the analyzed literature, it can be concluded that mechanisms of cariogenic pathogen inhibition using probiotics are still not well understood. The new research trend is based on application of probiotics which can naturally displace cariogenic bacteria in the oral cavity and influence oral health in adults and children. The results of studies also confirmed the beneficial role of probiotics in reduction of the bacterial population in periodontitis and halitosis. Long-term observation and a properly designed study protocol will allow us to answer many questions concerning substitution of one strain of bacteria by another.

Concurrent peritonsillar abscess and poststreptococcal reactive arthritis complicating acute streptococcal tonsillitis in a young healthy adult: a case report.

BACKGROUND: Streptococcus pyogenes is responsible for 5-15% and 20-30% of acute pharyngitis/tonsillitis in adults and children, respectively. It not only causes acute illness but also can give rise to local suppurative complications such as peritonsillar abscess as well as trigger the postinfectious syndromes of glomerulonephritis, acute rheumatic fever and poststreptococcal reactive arthritis. Here, we report a case of a young healthy adult in whom both peritonsillar abscess and poststreptococcal reactive arthritis developed as a complication of acute streptococcal tonsillitis. To the best of our knowledge, such a coincidence of poststreptococcal sequelae has not been reported previously. CASE PRESENTATION: A 32-year-old previously healthy woman was diagnosed with acute tonsillitis by her family doctor and treated empirically with amoxicillin/clavulanic acid (875/125 mg) twice daily for 5 days. Four days after completing antibiotic therapy, peritonsillar abscess of left tonsil developed. Needle aspiration followed by incision and drainage were performed by otolaryngologist at the Emergency Department. Next, the patient was discharged home on a 10-day course of cefuroxime and metronidazole. The symptoms of peritonsillar abscess were subsiding during treatment, however on the last day of antibiotic therapy, swelling and pain of the left ankle appeared. Five days later the patient was consulted by rheumatologist. Cultures of throat swabs and abscess aspirate collected 2 weeks before revealed the presence of Streptococcus pyogenes. Antistreptolysin O (ASO) titer was evaluated and proved to be 412 IU/ml (normal 0-200 IU/ml). The level of C-reactive protein was 13,0 mg/L (normal <5,0 mg/L). There was no known cardiac involvement. Poststreptococcal reactive arthritis was diagnosed. Left ankle arthralgia persisted for about 5-6 weeks. Six months after the presentation at the Emergency Department, the patient was well, with ASO titer reaching 262 IU/ml. CONCLUSIONS: Clinicians should be aware that appropriate choice of antibiotic, proper dose as well as duration of therapy of acute GAS pharyngitis/tonsillitis are crucial to prevent poststreptococcal sequelae.

Empirical validation of Polish guidelines for the management of acute streptococcal pharyngitis in children.

OBJECTIVE: Group A Streptococcus (GAS) pharyngitis is currently the only commonly occurring form of acute pharyngitis for which antibiotic therapy is definitely indicated. Polish guidelines advocate the use of modified Centor score (MCS) to assess the probability of GAS pharyngitis. They advise performing throat culture or rapid antigen detection test (RADT) in children with score 2-3 in MCS and treating with antibiotic only those in whom GAS was detected. Negative RADT results should be confirmed by culture. In children with score 4, the guidelines allow to introduce empiric antibiotic therapy. Phenoxymethyl penicillin is recommended as a drug of choice to treat GAS pharyngitis. The aim of our study was to evaluate the accuracy of strategy recommended by Polish guidelines in identifying those children with acute pharyngitis who require antibiotic treatment. Hence, diagnostic values of score 4 in MCS and RADT were assessed using throat culture as a reference standard. Phenoxymethyl penicillin efficacy in GAS eradication and prevention of post-streptococcal complications were estimated as well. METHODS: Ninety children between 2 and 15 years of age with acute pharyngitis symptoms suggesting GAS etiology (MCS ≥ 2), participated in our study. At the initial visit MCS was evaluated and two throat swabs were collected to perform RADT and culture. In children with GAS pharyngitis treated with penicillin, microbiological cure was assessed by performing two control throat cultures. Next, children were under observation for 3 months. RESULTS: Positive predictive value of score 4 in MCS turned out to be 48.05% (95% CI: 36.5-59.7%). RADT sensitivity, specificity and diagnostic accuracy proved to be 100%, 96%, and 98%, respectively. GAS eradication rate in children treated with penicillin turned out to be 92.5%. No post-streptococcal sequelae occurred in any child in 3-month observation. CONCLUSIONS: Empiric antibiotic therapy in children with score 4 in MCS will result in significant overtreatment of those with nonstreptococcal pharyngitis. New generation RADT diagnostic value in GAS detection proved to be equivalent to that of culture, which obviates the need of backup culture in children with negative RADT results. Phenoxymethyl penicillin revealed high eradication efficacy and proved to prevent post-streptococcal sequelae in children with acute GAS pharyngitis.

Management of acute streptococcal pharyngitis: still the subject of controversy.

Although most cases of acute pharyngitis are viral in origin, antibiotics are overused in its treatment. Streptococcus pyogenes (group A streptococcus, GAS), the principal bacterial pathogen of acute sore throat, is responsible for merely 5-30% of cases. Moreover, GAS pharyngitis is currently the only commonly occurring form of acute pharyngitis for which antibiotic therapy is definitely indicated. Therefore the differentiation between GAS pharyngitis and that of viral etiology is crucial. Accordingly, scientific societies as well as respected advisory bodies in Europe and North America, issued guidelines for the management of acute pharyngitis with the aim of minimizing unnecessary antibiotic prescriptions in its treatment. The aim of this review work is to confront the state of the art in acute GAS pharyngitis diagnosis and treatment with different approaches to its management represented by current European and North American guidelines. Although based on scientific evidence, international guidelines differ substantially in opinions whether GAS pharyngitis diagnosis should be based on microbiological testing, clinical algorithm or a combination of both. On the other hand, some European guidelines consider GAS pharyngitis to be a mild, self-limiting disease that does not require a specific diagnosis or antimicrobial treatment except in high-risk patients. There is an agreement among guidelines that if antibiotic therapy is indicated, phenoxymethyl penicillin should be the drug of choice to treat GAS pharyngitis.

[Utility of clinical differentiation and microbiological examination for appropriate antibiotic therapy of acute pharyngotonsillitis in children]. / Ocena przydatnosci parametrów klinicznych oraz badania mikrobiologicznego do optymalizacji antybiotykoterapii ostrego zapalenia gardia i migdalków podniebiennych u dzieci.

UNLABELLED: The differentiation between streptococcal pharyngotonsillitis and that of viral etiology is crucial, since it allows to discriminate patients who need to be treated with the use of antibiotics from those who only need symptomatic treatment. For that purpose, Centor/Mclssac's scale, based on the clinical examination and a history of a patient, is suggested by "Polish Recommendations for the diagnosis and treatment of community-acquired respiratory tract infections 2010". In patients who have obtained two or more points according to this scale, a definitive diagnosis of streptococcal pharyngotonsillitis depends on the identification of the bacterium, which can be performed by throat swab culture or by rapid antigen detection test. The aim of the study was to evaluate utility of clinical differentiation, rapid antigen detection test and throat swab culture for appropriate antibiotic therapy of acute pharyngotonsillitis in children. To the best of our knowledge, this is the first survey of this type carried out in Poland. MATERIAL AND METHODS: 42 children in the age of 3-14 years with acute pharyngotonsillitis who obtained at least two points in Centor/ Mclssac's scale took part in our study. Two throat swabs were obtained from every child, one of them was used to perform rapid test, and the second one was cultured on Columbia blood agar. Grey-white colonies with beta-hemolysis were classified to the groups A, B, C, D, F and G according to Lancefield with the use of bacitracin discs and latex agglutination test. RESULTS: 50% of children obtained 4 points and 40.5% of them received 5 points in Centor/Mclssac's scale. QuickView+ Strep A test turned up to be consistent with culture in 97.6% of cases. On the grounds of rapid test and culture results patients were subdivided into 3 groups: I--infected with S. pyogenes (50.0%), II--infected with other groups of beta-hemolytic streptococci (9.5%), III--infected with viruses (40.5%). CONCLUSIONS: It is impossible to recognize streptococcal pharyngitis only on clinical grounds, the diagnosis must be confirmed by laboratory methods. In children with high risk of streptococcal pharyngitis QuickView+Strep A test possesses diagnostic value comparable to culture in the detection of S. pyogenes, however negative rapid test results in such patients should be confirmed by culture. Antibiotic therapy should be initiated on the base of microbiological examination.

[Rational antibiotic therapy of acute upper respiratory tract infections]. / Racjonalna antybiotykoterapia ostrych zapalen górnych dróg oddechowych.

Overuse of antibiotics is a major cause of bacterial resistance. Inappropriate and unnecessary antibiotic therapies are particularly frequent in the treatment of respiratory tract infections, which are viral in origin in about 80% of cases. This review work presents the principles of rational antibiotic therapy of acute upper respiratory tract infections, namely pharyngotonsillitis, otitis media and sinusitis according to Polish Recommendations 2010.

Intestinal colonization with Oxalobacter formigenes and its relation to urinary oxalate excretion in pediatric patients with idiopathic calcium urolithiasis.

BACKGROUND AND AIM: Oxalobacter formigenes is an intestinal bacterium that utilizes oxalate as the only source of energy. It has been suggested that the lack of colonization with this organism may be a risk factor for calcium oxalate urolithiasis. Because this problem was not investigated in pediatric stone formers, we decided to assess it in our patients. METHODS: The presence of O. formigenes in stool samples of 76 children and adolescents (aged 4.1-18 years) with idiopathic calcium urolithiasis (36 with chemically confirmed calcium oxalate stones and 40 children with a strong clinical suspicion of this type of urolithiasis) was assessed using PCR method. Simultaneously, urinary oxalate excretion was measured in this group. Fifty healthy, age- and sex-matched subjects served as controls. RESULTS: O. formigenes was found in 21/76 patients (27.6%). In controls, frequency of colonization was similar (26%). The median 24h urinary oxalate excretion in patients colonized with O. formigenes was significantly lower in comparison with non-colonized patients, 0.319 (range 0.141-0.546) and 0.437 (range 0.198-0.967) mmol/1.73 m(2)/24h, respectively. CONCLUSIONS: Higher urinary oxalate excretion in children with calcium urolithiasis may be a result of the absence of O. formigenes. The reasons for similarly low intestinal colonization with this bacterium in normal subjects and stone formers remain speculative. Thus, further studies are necessary to clarify this issue.

Yeast holoenzyme of protein kinase CK2 requires both beta and beta' regulatory subunits for its activity.

Protein kinase CK2 is a highly conserved Ser/Thr protein kinase that is ubiquitous among eucaryotic organisms and appears to play an important role in many cellular functions. This enzyme in yeast has a tetrameric structure composed of two catalytic (alpha and/or alpha') subunits and two regulatory beta and beta' subunits. Previously, we have reported isolation from yeast cells four active forms of CK2, composed of alphaalpha'betabeta', alpha2betabeta', alpha'2betabeta' and a free alpha'-catalytic subunit. Now, we report that in Saccharomyces cerevisiae CK2 holoenzyme regulatory beta subunit cannot substitute other beta' subunit and only both of them can form fully active enzymatic unit. We have examined the subunit composition of tetrameric complexes of yeast CK2 by transformation of yeast strains containing single deletion of the beta or beta' regulatory subunits with vectors carrying lacking CKB1 or CKB2 genes. CK2 holoenzyme activity was restored only in cases when both of them were present in the cell. Additional, co-immunoprecypitation experiments show that polyadenylation factor Fip1 interacts with catalytic alpha subunits of CK2 and interaction with beta subunits in the holoenzyme decreases CK2 activity towards this protein substrate. These data may help to elucidate the role of yeast protein kinase CK2beta/beta' subunits in the regulation of holoenzyme assembly and phosphotransferase activity.

Yeast elf1 factor is phosphorylated and interacts with protein kinase CK2.

One of the biggest group of proteins influenced by protein kinase CK2 is formed by factors engaged in gene expression. Here we have reported recently identified yeast transcription elongation factor Elf1 as a new substrate for both monomeric and tetrameric forms of CK2. Elf1 serves as a substrate for both the recombinant CK2alpha' (K(m) 0.38 microM) and holoenzyme (K(m) 0.13 microM). By MALDI-MS we identified the two serine residues at positions 95 and 117 as the most probable in vitro phosphorylation sites. Coimmunoprecypitation experiments show that Elf1 interacts with catalytic (alpha and alpha') as well as regulatory (beta and beta') subunits of CK2. These data may help to elucidate the role of protein kinase CK2 and Elf1 in the regulation of transcription elongation.

[Etiological factors of vascular grafts infections and their resistance to antibiotics]. / Czynniki etiologiczne zakazen przeszczepów naczyniowych i ocena ich lekoopornosci.

THE AIM: of our study was the identification of microorganisms causing vascular graft infections and the evaluation of their antimicrobial susceptibility. MATERIAL AND METHODS: 25 patients with infected vascular graft, took part in our research. In 19 patients late type of infection was recognized, in 6 the infection was qualified as early. Purulent discharge obtained from the fistula was inoculated on the bacteriological media. Antimicrobial susceptibility was assessed by disc-diffusion method. RESULTS: Staphylococcus aureus and Pseudomonas aeruginosa proved to be the most frequently isolated microorganisms. Mixed infection, caused by two distinct bacteria, occurred in 5 patients; in all cases one species belonged to Gram-positive, and the second one to Gram-negative bacteria. In 50% of patients with early type infection different species of Gram-negative rods were present, in 37,5% of them S. aureus and S. epidermidis were isolated. In late type infection Gram-negative rods were isolated from 54,5% of patients and Gram-positive bacteria from 31,5% of patients. The most frequently isolated species appeared to be Pseudomonas aeruginosa. The isolated species of bacteria varied depending on the degree of infection (according to Shilagy and Samson). CONCLUSIONS: A diversity of isolated species, the presence of mixed infections and resistance patterns typical for hospital flora among bacteria infecting vascular grafts cause that antibiotic therapy should always be based on the results of microbiological examination.

Chlamydia pneumoniae infection in abdominal aortic aneurysm (AAA) patients and its clinical impact.

The aim of our study was to assess the presence of Chlmydia pneumoniae infection in AAA patients and to evaluate its association with clinical symptoms and histological signs of inflammation in the aortal wall. Fifty-two AAA patients participated in the research. Thirty healthy controls took part in serological examination. C. pneumoniae was detected by PCR and immunofluorescence in situ reaction in aorta samples of 84.6% and 86.54% of the patients, respectively. Serological markers of chronic C. pneumoniae infection were detected in 86.5% of AAA patients and in 33.3% of healthy controls. High titers of specific IgG and IgA were found in 37.8% of AAA patients with serologically defined chronic infection. All patients in "high serology" group had symptomatic aneurysm and inflammatory infiltrations in their aortal wall samples. AAA patients infected with C. pneumoniae are not a homogenous group. "High serology" group is much more prone to have symptomatic aneurysm than the remaining of AAA patients. Serology can be very useful in predicting the risk of AAA rupture. Inter-laboratory standardization of direct and indirect detection methods of C. pneumoniae infection is required to elucidate the role of these bacteria in AAA development.

Evaluation of an anti-chlamydial antibiotic therapy influence on asthma patients.

Chlamydia pneumoniae is one of the most frequent pathogens causing airways infections. Contribution of chronic chlamydial infection to the following diseases: asthma, POChP, coronary heart disease, abdominal aortic aneurysm, is particularly interesting. The connection between such infection and bronchial asthma was described in the literature in 1991. C. pneumoniae often causes asthma exacerbation; it is suggested that it also may be an etiologic factor of the disease. In a group of 55 subjects with chronic, stable bronchial asthma treated in the Pulmonary Department, serologic characteristic of C. pneumoniae infection was found in 34 patients (61,8%). Thirteen of these subjects agreed to participate in the study. They were divided into two groups; placebo was administered to the first one and azithromycin in a dose of 1000 mg once a week--to the other one. The research was conducted using the double blind trial method. Anti-chlamydial antibody level was evaluated before and after treatment. Spirometry tests as well as subjective estimation of physical fitness and dyspnoea degree were also determined. In comparison with 'the placebo group', statistically significant improvement in respiratory parameters 'in the treated group' was not ascertained.

Use of secretin in the treatment of childhood autism.

The paper presents current views concerning childhood autism. The authors present the concepts of etiology of this disorder, emphasizing the role of negative psychical stimuli in early childhood and the role of mother's contact with the child. Organic factors, including genetic background, developmental abnormalities of the nervous system, teratogenic factors and perinatal traumas are also taken into consideration. The role of metabolic factors and enterohormones, particularly those belonging to the secretin group and their effect on the function of the gastrointestinal tract and central nervous system is emphasized. We discuss signs which may be indicative of first symptoms of autism in different age groups. A typical symptom of autism is no development of speech, observed from infancy, taking the form of complete mutism at later stages. It has been emphasized that most pathologic symptoms result from altered perception of external stimuli, which arouse fear and anxiety. Autistic patients may suffer from gastrointestinal tract disturbances such as abdominal pains and diarrhea. Methods used hitherto in the therapy of childhood autism, mainly by psychologists and psychiatrists, as well as some attempts of pharmacological treatment, are presented. The structure and function of secretin, as well as its effects on the motor and secretory function of the stomach and the exocrine function of the pancreas are discussed. The role of secretin in diagnostic tests, among others in the diagnosis of gastrinoma, is emphasized. We also present the history of the application of secretin in the therapy of childhood autism.

Immunofluorescence in situ and the serologic indices of Chlamydia pneumoniae infection in patients with an abdominal aortic aneurysm.

The aim of the study was to evaluate the frequency of Chlamydia pneumoniae infection in patients with abdominal aortic aneurysm (AAA) using selected methods. The histological specimens of aneurysm wall were evaluated, the method of immunofluorescence was used to reveal the antigen in the wall of AAA and the titers of specific antibodies of IgG, IgM and IgA classes in blood plasma were marked. Atherosclerotic changes in the aneurysm wall were found in all patients. In 20(87%) patients the C. pneumoniae antigen was seen in the wall of abdominal aneurysm using the indirect immunofluorescence method. A significant relation between the method of direct C. pneumoniae diagnosis, aneurysm symptoms and histologically detected inflammation in its wall was confirmed. Serologic markers of the chronic C. pneumoniae infection were seen in 20(87%) out of 23 patients and in 6(30%) out of 20 subjects of the control group and this difference was statistically significant. It was observed, that all patients with serologic indices of active C. pneumoniae infection, had symptomatic aneurysm. The presence of Chlamydia pneumoniae in the wall of AAA as well as the occurrence of serologic indices of the chronic infection in these patients can confirm the hypothesis of the relation between the infection with this microorganism and the development of the disease.