Antibacterial activity of Hypericum perforatum L. (St. John's wort) extracts against Gram-positive bacteria and characterisation of its secondary metabolites.

This study aimed to determine the secondary metabolite profiles and antibacterial activity of H. perforatum L extracts against Gram-positive clinical isolates. The plant materials (Sample A and Sample B) were macerated with n-hexane, ethyl acetate and methanol (MeOH). The antibacterial activitiy of plant extracts and routinely used antibiotics were tested against Gram-positive bacteria. The secondary metabolite profiles of Sample A were determined by LC-Q-TOF-MS. The MIC values for n-hexane and ethyl acetate extracts of Sample A were lower than the susceptibility breakpoints of most broad-spectrum antibiotics (e.g. vancomycin, teicoplanin and linezolid) in a certain proportion of Gram-positive bacteria. The n-hexane extract of Sample A showed good antibacterial activity with MICs lower than the susceptibility breakpoint of teicoplanin in 58% of coagulase-negative staphylococci. The n-hexane and ethyl acetate extracts of Sample A had rich phloroglucinol constituents. The n-hexane and ethyl acetate extracts of Sample A could be alternative antibacterial agents against Gram-positive bacteria.

Distribution of spoligotyping defined genotypic lineages among drug-resistant Mycobacterium tuberculosis complex clinical isolates in Ankara, Turkey.

BACKGROUND: Investigation of genetic heterogeneity and spoligotype-defined lineages of drug-resistant Mycobacterium tuberculosis clinical isolates collected during a three-year period in two university hospitals and National Tuberculosis Reference and Research Laboratory in Ankara, Turkey. METHODS AND FINDINGS: A total of 95 drug-resistant M. tuberculosis isolates collected from three different centers were included in this study. Susceptibility testing of the isolates to four major antituberculous drugs was performed using proportion method on Löwenstein-Jensen medium and BACTEC 460-TB system. All clinical isolates were typed by using spoligotyping and IS6110-restriction fragment length polymorphism (RFLP) methods. Seventy-three of the 95 (76.8%) drug resistant M. tuberculosis isolates were isoniazid-resistant, 45 (47.4%) were rifampicin-resistant, 32 (33.7%) were streptomycin-resistant and 31 (32.6%) were ethambutol-resistant. The proportion of multidrug-resistant isolates (MDR) was 42.1%. By using spoligotyping, 35 distinct patterns were observed; 75 clinical isolates were grouped in 15 clusters (clustering rate of 79%) and 20 isolates displayed unique patterns. Five of these 20 unique patterns corresponded to orphan patterns in the SITVIT2 database, while 4 shared types containing 8 isolates were newly created. The most prevalent M. tuberculosis lineages were: Haarlem (23/95, 24.2%), ill-defined T superfamily (22/95, 23.2%), the Turkey family (19/95, 20%; previously designated as LAM7-TUR), Beijing (6/95, 6.3%), and Latin-America & Mediterranean (LAM, 5/95 or 5.3%), followed by Manu (3/95, 3.2%) and S (1/95, 1%) lineages. Four of the six Beijing family isolates (66.7%) were MDR. A combination of IS6110-RFLP and spoligotyping reduced the clustering rate from 79% to 11.5% among the drug resistant isolates. CONCLUSIONS: The results obtained showed that ill-defined T, Haarlem, the Turkey family (previously designated as LAM7-TUR family with high phylogeographical specifity for Turkey), Beijing and LAM were predominant lineages observed in almost 80% of the drug-Resistant M. tuberculosis complex clinical isolates in Ankara, Turkey.

Quantitative analysis of colonization with real-time PCR to identify the role of Oxalobacter formigenes in calcium oxalate urolithiasis.

The objective of the study was to quantitatively measure the number of Oxalobacter formigenes (O. formigenes) colonizations in the gastrointestinal tract in calcium oxalate-forming patients with real-time polymerase chain reaction (PCR). Calcium oxalate-forming patients (n: 27) were included in the study. Serum calcium, sodium, potassium, urea and creatinine levels, as well as 24 h urine levels of calcium and oxalate were measured. The numbers of O. formigenes colonies in stool samples were detected by real-time PCR. One or two metabolic abnormalities were detected in 15 of 27 patients. The O. formigenes levels in patients with metabolic disturbance were significantly decreased when compared to the patients with no metabolic abnormalities (p: 0.038). The undetectable levels of O. formigenes were encountered in one of five patients with hypercalciuria, in three of four patients with hyperoxaluria and in four of six patients with both hypercalciuria and hyperoxaluria. In nine patients with a history of stone recurrence, O. formigenes colonization was significantly lower than the patients with the first stone attack (p: 0.001). O. formigenes formation ceased or significantly diminished in patients with calcium oxalate stones with a coexistence of both hyperoxaluria and hypercalciuria. The measurement of O. formigenes colonies by real-time PCR seemed to be an inconvenient and expensive method. For this reason, the real-time PCR measurements can be spared for the patients with stone recurrences and with metabolic abnormalities like hypercalciuria and hyperoxaluria. The exact measurement of O. formigenes may also help more accurate programming of O. formigenes-based treatments.

Helicobacter pylori infection in mother and infant pairs in Anatolia.

BACKGROUND/AIMS: The aim of this prospective study was to determine the seroprevalence rates of Helicobacter pylori in mother and infant pairs and to discuss the possible fecal-oral transmission route of Helicobacter pylori infection in the early years of life. METHODS: Forty-eight mother-child pairs were followed for 12 months. Helicobacter pylori IgG and hepatitis A virus (HAV) IgG levels were measured in maternal sera, infant sera and breast-milk samples at birth and in breast-milk samples and infant sera at follow-up visits. RESULTS: At birth, the rate of Helicobacter pylori positivity was 81.25% in breast-milk and 95.8% in maternal and infant sera. Although there was a decrease in seropositivity in both baby sera and breast-milk at the age of nine months, an increase was observed in the 12th month. Hepatitis A virus IgG was measured to show whether Helicobacter pylori and hepatitis A virus use the same transmission routes. Hepatitis A was positive in all infants' sera, in 95.8% of mothers' sera, and in 68.75% of breast-milk samples. Seropositivity rates in infants whose mothers were seropositive for Helicobacter pylori and hepatitis A virus decreased gradually. There was an increase after the 9th month of life. CONCLUSIONS: Helicobacter pylori seroprevalence rates are high in Anatolia. It is possible that the decrease in breastfeeding with increased introduction of supplemental foods may lead to an increased risk of exposure to Helicobacter pylori.

Tuberculosis prevalence in forensic autopsies.

INTRODUCTION: According to the 2008 World Health Organization report, in 2006, 9.2 million new cases were determined, and 1.7 million people have lost their life due to tuberculosis (TB) in all around the world. In our country (Turkey), it is estimated that 35,000 to 40,000 people have TB disease annually. The Ministry of Health could just determine 18,500 of these cases, and only 6500 patient could be treated effectively. According to the Tuberculosis Dispensary records, the incidence for TB in Turkey is 28/100,000. MATERIALS AND METHODS: It is aimed to determine the infection with Mycobacterium tuberculosis using acidoresistant bacilli microscopy, TB culture, and histopathological methods in tissue samples that were obtained from lungs of forensic cases whose autopsies had been performed in Council of Forensic Medicine Ankara Department Morgue Specialized Committee. RESULTS: A total of 3 tissue samples that were obtained from lungs of randomized 302 cases, were positive for TB in Löwenstein-Jensen medium. Granuloma with caseating necrosis was found in histopathological examination and acidoresistant (+) bacilli (1+, 2+, and 2+, respectively) in microscopically analysis were also demonstrated in this 3 tissue samples. DISCUSSION: For this reason, we think that autopsy workers have to be careful about tuberculosis during their autopsy working.

Phosphorus-nitrogen compounds. 18. Syntheses, stereogenic properties, structural and electrochemical investigations, biological activities, and DNA interactions of new spirocyclic mono- and bisferrocenylphosphazene derivatives.

The reactions of hexachlorocyclotriphosphazatriene, N(3)P(3)Cl(6), with mono- (1 and 2) and bisferrocenyldiamines (3-5), FcCH(2)NH(CH(2))(n)NHR(1) (R(1) = H or FcCH(2)-), produce mono- (6 and 7) and spirocyclic bisferrocenylphosphazenes (8-10). The fully substituted phosphazenes (11-15 and 18-21) are obtained from the reactions of corresponding partly substituted phosphazenes (6-10) with excess pyrrolidine and NH(2)(CH(2))(3)ONa, respectively. The reactions of 6 with 1-aza-12-crown-4 afford geminal (16) and tris (17) crown ether-substituted phosphazenes. The structural investigations of the compounds have been verified by elemental analyses, mass spectrometry, Fourier transform IR, (1)H, (13)C, and (31)P NMR, and DEPT, COSY, HETCOR, and HMBC techniques. The crystal structures of 7, 10, 11, and 15 have been determined by X-ray crystallography. In 16 and 17, there are one and two stereogenic P atoms, respectively, and they are expected to be in enantiomeric mixtures. The structures of 18-21 look similar to a propeller. In 20 and 21, there are two stereogenic P atoms, and they exist as cis (meso; 20a and 21a) and trans (racemic; 20b and 21b) geometric isomers, according to the chiral solvating agent (S)-(+)-2,2,2-trifluoro-1-(9'-anthryl)ethanol experiments. Moreover, the compounds 18 and 19 have three stereogenic P atoms, and they exist as enantiomeric mixtures. Cyclic voltammetric investigations of compounds 6-21a reveal that ferrocene redox centers undergo oxidation concurrently at the same potential with basically reversible peaks, and these compounds appear to be quite robust electrochemically. The compounds 11-15 have been screened for antibacterial activity against gram positive and gram negative bacteria and for antifungal activity against yeast strains.The compounds 11, 12, 14, and 15 are evaluated for antituberculosis activity against reference strain Mycobacterium tuberculosis H37Rv (ATCC 27294). Interactions between compounds 11-15 and pBR322 plasmid DNA are studied by agarose gel electrophoresis. These compounds induce conformational changes in the DNA helix.

Polymerase chain reaction based diagnosis of primary gastric tuberculosis in an 80-year-old woman: a case report and review of the literature.

The patient had a two month history of gastrointestinal symptoms. Upper gastrointestinal endoscopy disclosed 5 mm nodular lesions were seen in the prepyloric area. On pathological examination, two granulomatous lesions were detected in biopsy specimen. Ehrlich Ziehl-Neelsen staining and cultures of the biopsy material were negative, but polymerase chain reaction (PCR) for Mycobacterium tuberculosis complex DNA was positive. Clinical diagnosis of primary gastric tuberculosis (PGTb) was supported by positive PCR assay and histopathological findings. After antituberculosis treatment, nodular lesions were not detected. The diagnosis of PGTb was confirmed definitively by the success of treatment and repeated endoscopic examination.

Genetic diversity of Mycobacterium tuberculosis isolates at the Military Medical Academy in Ankara, Turkey.

Genotyping of Mycobacterium tuberculosis isolates from infected individuals can play an important role in tracking the source of infection and unraveling the epidemiology of a tuberculosis pandemic. A total of 114 M. tuberculosis isolates were genotyped by spoligotyping and results were compared with an international spoligotype database (SpoIDB4). Twenty-one spoligotyping-defined clusters including 97 patients were established, and an additional 17 unique patterns were found. Ninety-eight (85.9%) isolates belonged to previously defined shared types (STs). The ST53 (ill-defined T1 superfamily, n=31), ST41 (LAM7-TUR family, n=9), ST118 (T undefined, n=8) and ST50 (Haarlem 3, n=6) were four major clusters of our isolates. After comparison with the international SpoIDB4 database, two new intrafile clusters, ST2136 and ST2139, were created and two new interfile clusters, ST2135 and ST2140, were defined. Eight (7%) of the 17 isolates with unique patterns were found to be orphans, whereas the STs of 9 isolates had previously been deposited in the international SpoIDB4 database. In addition, two isolates with an ST pattern characteristic of the Beijing family of M. tuberculosis were found. This study shows that, although ubiquitous spoligotypes are common, several spoligotypes specific to Turkey also exist. Thus, our study may help us to better understand the spread of M. tuberculosis genotypes to Turkey.

The predictive value of serum procalcitonin levels in adult patients with active pulmonary tuberculosis.

The aim of our prospective study was to evaluate the predictive value of serum procalcitonin (PCT) level in comparison with C-reactive protein level and erythrocyte sedimentation rate for the diagnosis of pulmonary tuberculosis (PTB) on admission and 6 months after the administration of anti-tuberculous chemotherapy (ATCT). Seventy-five adult male patients with active PTB who were mycobacteriologically diagnosed (smear and culture positivity) were examined in this study. As a control group, 75 healthy adult males were enrolled. The measured serum PCT levels were within the normal range both in healthy individuals and in patients 6 months after ATCT. Serum PCT levels had been slightly high on admission in patients with PTB in comparison with controls (P = 0.01) and patients who had ATCT (P = 0.001), and this difference was statistically significant, but the PCT levels of most cases with PTB (58.7%) were below the usual cut-off level (0.5 ng/mL). We conclude from this study that the serum PCT level was not a reliable indicator in the diagnosis of active PTB because of its low sensitivity (41.3%), and in most cases it was not capable of overcoming the cut-off level even if statistically meaningful results were obtained. The PCT test for the presumptive diagnosis of PTB cannot be substituted for microbiological, epidemiological, clinical and radiological data.

A case of tuberculous pyomyositis that caused a recurrent soft tissue lesion localized at the forearm.

We present the case of a 20-year-old male who had a non-traumatic soft tissue lesion (4 x 3 cm) with recurrent discharge at his right posteromedial antebrachial muscles; the patient underwent surgery twice, and antibiotic therapy was administered, but no cure was achieved with these treatments. The patient underwent surgery at our medical center. There was no history of pulmonary, gastrointestinal, or genitourinary tuberculosis (TB). Due to suspected pulmonary, genitourinary, and gastrointestinal TB, radiography and computed tomography scans were performed, and these studies disclosed no evidence of a primary origin. The erythrocyte sedimentation rate and the results of purified protein derivate testing were normal. We also detected submandibular lymphadenopathy (LAP) (2 x 3 cm) localized at a submandibular site in our patient 4 months after his first visit to our clinic. Smears were stained with Ehrlich Ziehl Neelsen (EZN) stain and culture were grown for Mycobacterium tuberculosis complex (MTC); the samples used for these assays had been obtained by incisional biopsy of the forearm lesion and by aspiration of the submandibular lymph node, and they were found to be MTC-positive. Then, a culture for MTC, derived from an induced sputum sample, was found to be positive, despite the negative results obtained with a sputum smear subjected to EZN staining. According to these results, the primary focus of the tuberculous pyomyositis and the submandibular LAP was the lungs. The lesion and submandibular LAP were both treated successfully by the administration of antituberculous chemotherapy.

[Detection of primary drug resistance rates of Mycobacterium tuberculosis complex strains isolated between 2002-2003 years and comparison with the data of 1998-2001 period]. / 2002-2003 yillari arasinda izole edilen Mycobacterium tuberculosis kompleks suslarinda primer ilaç direnç oranlarinin saptanmasi ve 1998-2001 verileriyle karsilastirilmasi.

In this retrospective study, it was aimed to evaluate the primary drug resistance rates of 151 Mycobacterium tuberculosis complex (MTC) strains against primary anti-tuberculosis (anti-TB) drugs, isolated from 2213 tuberculosis suspected patients between January 2002 to December 2003 in our hospital, and also to compare these results with the data obtained from our previous results between January 1998 to December 2001. It has been detected that there was a significant decrease in the susceptibility rates of MTC isolates to all anti-TB drugs both yearly from 1998 to 2003 (p < 0.001), and for the total of 1998 and 2003 years period (p < 0.029). When only the years 1998 and 2003 were taken into consideration, significant increases were found in any (one or more) combination of the drugs with isoniazid (INH), and only INH resistance rates (p < 0.007 and p < 0.033, respectively). Thus, naturally there was also an increase in resistance to total any drug resistance (p < 0.029). We suggest that strict measures such as "directly observed therapy" should be undertaken in order to prevent development of drug resistance particularly against INH. Regular and continuous screening of anti-TB drug resistance should be accomplished to survey the presence of drug resistant strains in the nation, to define the suitable drug regimens and to evaluate the quality of tuberculosis control programs.

The evaluation of FASTPlaqueTB test for the rapid diagnosis of tuberculosis.

FASTPlaqueTB (Biotec Laboratories Ltd., Ipswich, UK) is a rapid test which utilizes bacteriophage amplification technology for the detection of viable Mycobacterium tuberculosis in clinical specimens. We evaluated performance of the FASTPlaqueTB test by comparing with BACTEC 460 TB culture system (Becton Dickinson Co., Maryland, USA), polymerase chain reaction (PCR) and acid fast bacilli (AFB) smear methods. We investigated 192 sputum specimens collected from the patients suspected of having pulmonary TB by AFB smear, BACTEC 460 TB culture system, PCR and FASTPlaqueTB test. The sensitivity of AFB smear, PCR and FASTPlaqueTB test were 57.8%, 84.4% and 87.5% respectively when we accepted BACTEC 460 TB culture system as gold standard. We conclude that FASTPlaqueTB test has a good potential for rapid diagnosis of Mycobacterium tuberculosis as a result of the evaluation of these three tests by comparison to the BACTEC 460 TB culture system.

The evaluation of diagnostic methods for the detection of Helicobacter pylori in gastric biopsy specimens.

PCR is a rapid, sensitive and accurate method for the specific detection of Helicobacter pylori from gastric biopsy specimens. In our study, 104 gastric tissue specimens from symptomatic adult patients were examined by staining, culture, PCR and nested PCR methods for detection of H. pylori. According to our results, positivity was achieved in 24% (25/104) with Giemsa staining, 34% (36/104) with histopathology, 36% (38/104) with PCR and 41% (43/104) with nested PCR respectively, whereas H. pylori was isolated in only 33% (35/104) of the cultures on the biopsy specimens. Both the sensitivity and the positive predictive value of the nested PCR method were 100%, and both the specificity and negative predictive value were 98%. As a conclusion, our results suggest the nested PCR as a highly valuable method in the detection of H. pylori with a reasonably high sensitivity and specificity.