Effect of DNA damage of cumulus oophorus cells and lymphocytes analyzed by alkaline comet assay on oocyte quality and intracytoplasmic sperm injection success among patients with polycystic ovary syndrome.

AIM: Cumulus oophorus cells (COC) exhibit a close relationship with the oocytes. We aimed to determine the role of the DNA integrity of COC and lymphocytes on the oocyte and embryo quality and intracytoplasmic sperm injection (ICSI) success. METHODS: The COC obtained with the mechanical denudation of each oocyte and the lymphocytes obtained on oocyte retrieval day from 69 infertile polycystic ovary syndrome (PCOS) patients were used. The tail length, tail moment and tail DNA percentage were evaluated using an alkaline comet assay. The oocytes and embryos were graded. Clinical pregnancy was defined as the presence of a gestational sac with a beating heart. RESULTS: All of the DNA integrity parameters of the COC and lymphocytes were similar between the good and poor quality oocytes and between the good and poor quality embryos. There was no relationship between the DNA damage parameters and the fertilization and clinical pregnancy. The day 2 hormone levels, body mass index and age were negatively correlated with the DNA integrity parameters. No links were found between the embryo and oocyte scores and the DNA damage parameters. CONCLUSION: The DNA integrity of the COC and lymphocytes did not seem to be related to the oocyte and embryo quality and ICSI success.

Distribution of aluminum over different T-sites in ferrierite zeolites studied with aluminum valence to core X-ray emission spectroscopy.

The potential of valence to core Al X-ray emission spectroscopy to determine aluminum distribution in ferrierite zeolites was investigated. The recorded emission spectra of four samples prepared with different structure directing agents exhibit slight variations in the position of the main emission peak and the intensity of its low energy shoulder. Theoretical calculations indicate that an increased intensity of the Kßx shoulder in the Al emission spectra can be linked to a predominant occupation of the T3 site by a single aluminum atom. This study thus suggests that valence to core X-ray emission spectroscopy can be applied to help determine the occupation of aluminum at crystallographic T-sites in zeolites.

Effect of dietary probiotic and high stocking density on the performance, carcass yield, gut microflora, and stress indicators of broilers.

A study was carried out to evaluate the effect of dietary probiotic supplementation and stocking density on the performance, relative carcass yield, gut microflora, and stress markers of broilers. One-day-old Ross 308 male broiler chickens (n = 480) were allocated to 4 experimental groups for 42 d. Each treatment had 8 replicates of 15 chicks each. Two groups were subjected to a high stocking density (HSD) of 20 birds/m² and the other 2 groups were kept at low stocking density (LSD) of 10 birds/m². A basal diet supplemented with probiotic 1 and 0.5 g/kg of diet (in starter and finisher diets, respectively) was fed to 2 treatments, one with HSD and the other with LSD, thereby making a 2 × 2 factorial arrangement. There was no interaction between stocking density (LSD and HSD) and dietary probiotic (supplemented and unsupplemented) for all the variables. Feed intake and weight gain were significantly low and feed conversion ratio was poor in broilers at HSD. Dietary probiotic significantly enhanced the feed intake and weight gain in starter phase only. Dietary probiotic supplementation had no effect (P > 0.05) on total aerobs, Salmonella sp., and Lactobacilli populations in the intestines of broilers. However, HSD reduced the Lactobacilli population only (P < 0.05). Relative breast yields were significantly higher in broilers reared at LSD than HSD. Thigh meat yield was higher in broilers in HSD group compared to LSD. Dietary probiotic did not affect the relative carcass yield and weight of lymphoid organs. Serum malondialdehyde, corticosterone, nitric oxide, and plasma heterophil:lymphocyte ratio were not affected either by stocking density or dietary probiotic supplementation. In conclusion, HSD negatively affected the performance and intestinal Lactobacilli population of broilers only, whereas probiotic supplementation enhanced the performance of broilers during the starter phase only. Total aerobes, Salmonella, Lactobacilli carcass yield, and stress indicators of broilers were not affected by the dietary supplementation of probiotic under the conditions of the present study.

The influence of different sugammadex doses on neural tube development in early-stage chick embryos.

BACKGROUND: Sugammadex is a modified gamma-cyclodextrin that has been developed with the goal of reversing the steroidal neuromuscular blocking agents. The aim of the present study is to investigate the effects of different sugammadex doses on embryologic and neural tube development in an early-stage chick embryo model. METHODS: A total of 100 specific pathogen-free, fertilized domestic chicken eggs were randomly divided into five groups (n = 20, each), and placed in an automatic cycle incubator. The eggs in the "control (C)" group were incubated without administration of any drug till the end of the experiment. Sub-blastodermic administration of 0.9% NaCl as vehicle control (VC) and different doses of sugammadex solutions prepared with the latter [2 mg/mL (LD), 4 mg/mL (MD), 16 mg/mL (HD)] were performed at 30 hr of incubation. All embryos were removed from the eggs at 72 hr when they were expected to reach Hamburger-Hamilton (HH) stages 19-20, then they were fixed, and evaluated histo-morphologically. RESULTS: Embryonic development was not observed in 11 eggs (1 in C, 1 in VC; 3 in LD, 3 in MD, and 3 in HD). All the developed embryos were compatible with the HH stages 19-20. A neural tube closure defect was detected in one embryo in the HD group. No statistically significant difference was found between the groups in terms of embryonic and neural tube developments. CONCLUSIONS: No significant association was found between the drug and adverse outcomes; however, a trend with dosing was seen. Further studies are required before conclude on safety and extrapolate these results to human beings.

Frog saliva-induced toxic keratopathy: a case report.

Corneal toxicity due to exposure to many chemicals and medications have been described in the literature. We present a case of toxic keratopathy by corneal exposure to salivary secretion of a frog. A 40-year-old male patient reported a sudden splash of frog saliva in his right eye while he was examining it at a close distance. Corneal punctate epitheliopathy and stromal oedema and Descemet folds were the initial clinical findings, which completely recovered on the 2nd control day of topical dexamethasone and ofloxacin treatment. We aimed to show the toxic effects of animal-derived secretions on the cornea as a rare cause of toxic keratopathy.

The bird species of Kumasir lake (Kahramanmaras-Turkey) and a view of environmental ethics on sustainable wetland management.

Kumasir lake is located next to towns of Donuklu and Fatih, nine km west of Kahramanmaras city center the region of east Mediterranean, Turkey This lake is of crucial importance from the point of native and immigrant birds. We located 17 birdspecies in this area during our observations carried out in the spring and autumn of 2005-2006. These were Ciconia ciconia L., Anas platyrhynchos L., Accipiter nisus L., Accipiter brevipes L., Fulica atra L., Columba palumbus L., Merops apiaster L., Upupa epops L., Alauda arvensis L., Motacilla flava L., Turdus merula L., Acrocephalus scirpaceus L., Regulus regulus L., Garrulus glandarius L., Corvus corax L., Fringilla coelebs L., Hirundo rustica L.. Among observed 17 species; 6 of them were immigrant and remaining 11 of them were native birds. Kumasir lake is surrounded by wetland of Amik and Gavur lake. Since it was greatly dried, it was transformed to farmland. Consequently the birds lost most of theirnests and settlements. However not taken in the care of environmental ethic values, the wastewaters of the villages drain to lake reservoir; herbicides and insecticides used for agriculture are polluting the water reeds have been burned, the lake's reeds are getting dry by the irrigation for the farmland. So, the wetland ecosystem is being affected negatively by these factors. On the other hand, the birds are exposed to illegal and unlawful hunting. For this reasons, this lake must be taken into a management regime of sustainable wetland (protection profiting balance) and used techniques of participation planning via the process of sustainable natural resources and planning.

Postoperative endophthalmitis caused by Bacillus cereus and Chlamydia trachomatis.

PURPOSE: To describe the clinical manifestations and outcomes in 4 patients with endophthalmitis caused by Bacillus cereus and Chlamydia trachomatis. SETTING: Department of Ophthalmology, Ankara Training and Research Hospital, Ankara, Turkey. METHODS: Four patients who had cataract extraction and intraocular lens implantation with phacoemulsification at a secondary eye-care center presented with endophthalmitis. Cultures and direct fluorescein assay (DFA) were performed on vitreous aspirates from all patients. RESULTS: Cultures were positive for B cereus and DFAs were positive for C trachomatis in all patients. Despite timely intervention, at the end of follow-up, 1 patient had 20/200 visual acuity and another, counting fingers at 3 m. Phthisis bulbi developed in the 2 other patients. CONCLUSION: The course of infection with B cereus and C trachomatis poses a potential threat, especially because of the limited data on treatment of endophthalmitis secondary to C trachomatis.

Effects of hazelnut-enriched diet on plasma cholesterol and lipoprotein profiles in hypercholesterolemic adult men.

OBJECTIVE: Frequent consumption of nuts is associated with favorable plasma lipid profiles and reduced risk of coronary heart disease (CHD). This study was conducted to investigate the effects of hazelnut-enriched diet on plasma cholesterol and lipoprotein profiles in hypercholesterolemic adult men compared with baseline and control diet, and also to measure the anthropometric parameters, habitual physical activities, nutrient intake and endothelial function. SUBJECTS AND DESIGN: Fifteen hypercholesterolemic men aged 48+/-8 years were recruited voluntarily. A well-controlled, 2-period (P1 and P2) study design with a total of 8-week was implemented. In the P1, subjects consumed a control diet (low-fat, low-cholesterol and high-carbohydrate). During the P2, the control diet was supplemented with MUFA-rich hazelnut (40 g/day), which provided 11.6% of total energy content. Anthropometric parameters and habitual physical activities were recorded. Plasma total and HDL cholesterol, TAG, ApoA-1, Apo B, total homocysteine and glucose concentrations were measured. All parameters and measurements were obtained at baseline and end of each 4-week diet period. RESULTS: Body weights of subjects remained stable throughout the study. Compared with baseline, the hazelnut-enriched diet decreased (P<0.05) the concentrations of VLDL cholesterol, triacylglycerol, apolipoprotein B by 29.5, 31.8, and 9.2%, respectively, while increasing HDL cholesterol concentrations by 12.6%. Total/HDL cholesterol and LDL/HDL cholesterol ratios favorably decreased (P<0.05). Although insignificant there was a decreasing trend for the rest of parameters, particularly in total (5.2%) and LDL cholesterol (3.3%) in subjects consuming a hazelnut-enriched diet compared to that of the baseline. No changes were found in fasting levels of glucose, Apo A-1 and homocysteine between the control and hazelnut-enriched diets. CONCLUSIONS: This study demonstrated that a high-fat and high-MUFA-rich hazelnut diet was superior to a low-fat control diet because of favorable changes in plasma lipid profiles of hypercholesterolemic adult men and, thereby positively affecting the CHD risk profile. SPONSORSHIP: Funding provided by a grant from the Hazelnut Promotion Group, Giresun, Turkey.

Neutrophil volume distribution width as a new marker in detecting inflammatory bowel disease activation.

INTRODUCTION: We sought to investigate the value of neutrophil volume distribution width in detecting inflammatory bowel disease activation. METHODS: Patients with infection and accompanying inflammatory disease were excluded. All the patients were diagnosed and classified according to Porto criteria and Paris classification, respectively. Physician global assessment, pediatric Crohn's disease and pediatric ulcerative colitis activity indexes and fecal calprotectin were used to define disease activation. RESULTS: A total of 34 pediatric patients with Inflammatory bowel diseases (IBD) and 29 controls were enrolled in the study. Neutrophil volume distribution width (NVDW) was significantly higher in patients with IBD compared to healthy controls (P < 0.001). An increased NVDW level was observed in IBD patients in activation (22.42 ± 2.13) compared to those in remission (19.22 ± 1.63) (P < 0.001). There was no statistically significant difference between IBD patients in remission and healthy controls. The best cutoff of NVDW for prediction of disease activation in Crohn's disease and ulcerative colitis in this series was 20.39 with a sensitivity of 90.9% and a specificity of 75% (AUC: 0.852 CI: 0.698-1.000 P < 0.001) and 19.74 with a sensitivity of 92.9% and a specificity of 90.9% (AUC: 0.961, CI: 0.889-1.000, P < 0.001), respectively. CONCLUSIONS: As a quantitative, objective, and sensitive parameter, we believe that the NVDW has a potential to be an additional test detecting disease activation in IBD.

Evaluation of Pump Discontinuation and Associated Factors in the T1D Exchange Clinic Registry.

BACKGROUND: The objectives of this study were to examine factors associated with insulin pump discontinuation among children and adults followed longitudinally for 1 year in the multicenter T1D Exchange clinic registry, and to provide participant-reported reasons for stopping pump therapy. METHODS: We longitudinally followed 8935 participants of all ages using an insulin pump at the time of registry enrollment. Logistic regressions were used to identify demographic and clinical factors associated with pump discontinuation. Pump discontinuation was self-reported by participants on a first annual follow-up survey. RESULTS: The overall frequency of pump discontinuation was 3%. Discontinuation was higher in adolescents (4%) and young adults (4%) than in younger children (3%) or older adults (1%). In multivariate analysis of children between 6 and <13 and 13 and <18 years, participants who discontinued pump use were more likely to have higher HbA1c levels at baseline (adjusted P < .001 for both). The top participant-reported reasons for discontinuing the pump included problems with wearability (57%), disliking the pump or feeling anxious (44%), and problems with glycemic control (30%). CONCLUSIONS: In T1D Exchange registry participants, insulin pump discontinuation is uncommon, but more prevalent among adolescents and young adults, and youth with poor glycemic control. Given the known benefits of pump therapy, these populations should be targeted for support and education on troubleshooting pump use. Common reasons for discontinuation should also be considered in future device design and technological improvement.

Genomic backgrounds of drug-resistant Streptococcus pneumoniae in Ankara, Turkey: Identification of emerging new clones.

Streptococcus pneumoniae exhibiting decreased susceptibility to penicillin are isolated with an increasing prevalence in Turkey during the last decade. This study was undertaken to investigate the molecular epidemiology of non-penicillin-susceptible pneumococci isolated in Ankara, Turkey. Among a population of 246 pneumococci, 90 pneumococci with penicillin MIC > or = 0.1 microg/ml were serotyped, genotyped by pulsed-field gel electrophoresis (PFGE), and sequence typed by multilocus sequence typing (MLST). The overall resistance to penicillin, cefotaxime, erythromycin, clindamycin, chloramphenicol, tetracycline, rifampicin, ciprofloxacin, and vancomycin were 36.6%, 4%, 27.6%, 10.9%, 5.3%, 22.4%, 4.5%, 2%, and 0, respectively. The most frequent serotypes were 14, 23B, 9V, 19F, 19A, and 23F. PFGE types represented 17 genetic clusters. PFGE and MLST data revealed that there were isolates identical or closely related to the Spain(9V)-3 ST 156 clone, Portugal(19F)- 21 ST 177 clone, and Spain(23F)-1 ST81 clone. Eleven serotype 14 isolates with emerging resistance to penicillin belonged to the ST 230 complex, a predominantly susceptible clone. Serotype 19A, 19F, and 7F variants of the ST 230 clone were also identified in the study population. Eight serotype 23B isolates with a new ST 1349 (18-13-8-6-3-6-8) created another clone with no relation to the currently defined international clones. Although the pandemic clones Spain(9V)-3, Portugal1(9F)-21, and Spain(23F)-1 are present in our region, the emergence of a new 23B clone with a unique ST and the emergence of resistance in the ST230 clone, has presumably contributed to the increase in the prevalence of drug-resistant pneumococci in Turkey.

Prevalence of erm genes encoding macrolide-lincosamide-streptogramin (MLS) resistance among clinical isolates of Staphylococcus aureus in a Turkish university hospital.

This study investigated the prevalence of the erm(A), erm(B) and erm(C) genes among 122 MLS-resistant clinical isolates of Staphylococcus aureus from a Turkish university hospital. Of these isolates, 44 were inducibly resistant and 78 were constitutively resistant. The presence of one or more erm genes was demonstrated in 114 isolates; the erm(C) gene was detected in 97 isolates, and the erm(A) gene was detected in 96 isolates. Seventy-eight isolates harboured both erm(A) and erm(C). The combination of erm(A), erm(B) and erm(C) genes was detected in only one isolate.

[Vascular access outcome in patients aged over 75 years]. / Resultados del acceso vascular en mayores de 75 anos.

UNLABELLED: We report a retrospective study on the results of 132 native fistulas, 12 grafts and 27 tunneled catheters followed during 30 months in 144 patients on hemodialysis. The results were compared according to patient age: 75 years or over (n = 58, 80.3 +/- 3.5) vs below 75 years (n = 86, 59.5 +/- 13.3). Gender, presence of diabetes and type of fistula were also included in the analysis. RESULTS: There were no statistically significant differences between both groups in the use of tunneled catheters or grafts (8.6% vs 5.8% y 5.2% vs 10.5% respectively), primary failure of native fistulas (7.1% in those aged 75 years or over vs 25.5% in patients below 75 years), rate of thrombosis (0.03 vs 0.09/patient year at risk respectively) or number of percutaneous or surgical procedures in order to maintain the fistula patency (0.11 vs 0.16/patient year at risk respectively). At the same time no differences were seen in the primary, primary assisted and secondary patency of the native fistulas. The mean age of the patients when the first access fistula was created was different according to the area of surgery (74.9 +/- 9.3 for the elbow vs 64.9 +/- 16.2 years for the forearm, p < 0.005). Diabetes was an unfavourable factor for primary (HR Cox 2.08, p < 0.05) or secondary (Log Rank, p <0.05) patency. CONCLUSION: The vascular access for hemodialysis in elderly patients presents a similar evolution to that seen in younger populations if the access creation is based on an exhaustive study, including ecodoppler of the vascular map and the use of more proximal fistulas if necessary. Therefore the more frequent use of grafts or catheters in elderly patients is not justified.

Evaluation of a new rapid lateral flow chromatography test for the diagnosis of Helicobacter pylori.

OBJECTIVE: The rapid, simple and non-invasive diagnosis of Helicobacter pylori (H. pylori) infection is important in implementing chemotherapy in appropriate manner, and in assessing persistent H. pylori infection after eradication therapy. The ImmunoCard STAT! HpSA kit (Meridian Bioscience, Europe) is a lateral flow chromatography test which utilizes a monoclonal anti-H. pylori antibody. In this study, we investigated the usefulness of the ImmunoCard STAT! HpSA test before and after eradication therapy on patients referred to undergo upper gastrointestinal endoscopy. METHODS: Sixty-five consecutive patients who were referred to undergo upper gastrointestinal endoscopy at the Department of Gastroenterology, Karadeniz Technical University Medical School, Turkey between February and August 2005 were included in this study. The ImmunoCard STAT! HpSA was compared with 4 invasive tests (histology, gram staining, rapid urease test, and culture). The reference method was defined as positive when 2 of the 4 invasive tests were positive. A negative H. pylori status was considered when all 4 tests present concordant negative results. RESULTS: Overall, the ImmunoCard STAT! HpSA test had 77.8% sensitivity, 79.3% specificity, 82.4% positive predictive value (PPV) and 74.2% negative predictive value (NPV) in all patients. With regard to pre-treatment values, the sensitivity was 70.6%, specificity 70.6%, PPV 100% and NPV 100% while on post-treatment group the sensitivity was 84.2%, specificity 64.7%, PPV 72.7% and NPV 78.6%. CONCLUSION: Our results indicate that the ImmunoCard STAT! HpSA test is a rapid, simple, and helpful procedure not only to determine H. pylori infection but also to assess the success of eradication therapy.

Inflammasome activity is essential for one kidney/deoxycorticosterone acetate/salt-induced hypertension in mice.

BACKGROUND AND PURPOSE: Inflammasomes are multimeric complexes that facilitate caspase-1-mediated processing of the pro-inflammatory cytokines IL-1ß and IL-18. Clinical hypertension is associated with renal inflammation and elevated circulating levels of IL-1ß and IL-18. Therefore, we investigated whether hypertension in mice is associated with increased expression and/or activation of the inflammasome in the kidney, and if inhibition of inflammasome activity reduces BP, markers of renal inflammation and fibrosis. EXPERIMENTAL APPROACH: Wild-type and inflammasome-deficient ASC(-/-) mice were uninephrectomized and received deoxycorticosterone acetate and saline to drink (1K/DOCA/salt). Control mice were uninephrectomized but received a placebo pellet and water. BP was measured by tail cuff; renal expression of inflammasome subunits and inflammatory markers was measured by real-time PCR and immunoblotting; macrophage and collagen accumulation was assessed by immunohistochemistry. KEY RESULTS: 1K/DOCA/salt-induced hypertension in mice was associated with increased renal mRNA expression of inflammasome subunits NLRP3, ASC and pro-caspase-1, and the cytokine, pro-IL-1ß, as well as protein levels of active caspase-1 and mature IL-1ß. Following treatment with 1K/DOCA/salt, ASC(-/-) mice displayed blunted pressor responses and were also protected from increases in renal expression of IL-6, IL-17A, CCL2, ICAM-1 and VCAM-1, and accumulation of macrophages and collagen. Finally, treatment with a novel inflammasome inhibitor, MCC950, reversed hypertension in 1K/DOCA/salt-treated mice. CONCLUSIONS AND IMPLICATIONS: Renal inflammation, fibrosis and elevated BP induced by 1K/DOCA/salt treatment are dependent on inflammasome activity, highlighting the inflammasome/IL-1ß pathway as a potential therapeutic target in hypertension.

Congenital keratoglobus with multiple cardiac anomalies: a case presentation and literature review.

Keratoglobus is a rare condition of bilateral corneal ectasia, which results in high myopia, irregular astigmatism, scarring, and rarely spontaneous globe rupture. Globoid protrusion of a clear, diffusely thin cornea is the pathology. The congenital form has been associated with blue sclera in which there is a systemic connective tissue disorder with abnormal collagen synthesis like Ehlers-Danlos syndrome, Marfan syndrome, and osteogenesis imperfecta. Some concomitant abnormalities reported with kertoglobus include joint hypermobility, dental and skeletal abnormalities, osteal fragility, and deafness. Acquired forms have been reported to be associated with vernal keratoconjunctivitis and thyroid ophthalmopathy. We report the case of a 16-year-old boy with keratoglobus who presented with a history of photophobia and a low vision in both eyes since birth. He has been followed up by our pediatric cardiology department due to multiple cardiac anomalies. He had hypermobility of large joints, easy bruising, thin and hyperextensible skin with visible veins, which were also described in his elder brother. We aimed to discuss the etiology and the association of keratoglobus with some systemic abnormalities caused by collogen tissue disturbance, and make a brief review about the recent literature concerning the management of keratoglobus patients.

Resveratrol Pretreatment Ameliorates TNBS Colitis in Rats.

Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory disease in humans constituting a major health concern today whose prevalence has been increasing over the world. Production of reactive oxygen species (ROS) and disturbed capacity of antioxidant defense in IBD subjects have been reported. Antioxidants may play a significant role in IBD treatment. This study aimed at evaluating ameliorative effects of intraperitoneal resveratrol pretreatment on trinitrobenzene sulphonic acid (TNBS)-induced colitis in rats. Thirty five Wistar-Albino female rats were divided equally into five groups. Inflammation was induced by the intrarectal administration of TNBS under anesthesia. Intraperitoneal administration of resveratrol (RSV) at a concentration of 10mg/kg/day for 5 days before the induction of colitis significantly reduced microscopy score and malondialdehyde (MDA) levels and increased glutathione peroxidase (GSH Px) activity compared to TNBS and vehicle groups. Also an insignificant increase in catalase (CAT) activity was observed in the RSV treated group compared to TNBS and vehicle groups. In this paper, the most recent patent on the identification and treatment of IBD was indicated. In conclusion, antioxidant RSV proved to have a beneficial effect on TNBS colitis in rats. In light of these advantageous results, the RSV can be considered as adjuvant agent in IBD treatments.

Location of Ge and extra-framework species in the zeolite ITQ-24.

The germanosilicate ITQ-24 (IWR framework type) was synthesized in fluoride medium using 1,3,5-tris(1,2-dimethylimidazolium) benzene as the structure directing agent (SDA). A structure analysis of the as-synthesized ITQ-24 material using synchrotron powder diffraction data and difference electron density calculations have allowed the fluoride ions and the germanium atoms to be located and the conformation of the SDA to be determined. The benzyl ring is perpendicular to the b axis with the three imidazolium moieties forming a "T-shaped" arrangement. Ge atoms replace some of the Si in the double-4-ring (d4r) and in one of the single-4-rings (s4r). The other s4r contains only Si. Fluoride ions are in the d4r units. Initially, the space group Cmmm (highest possible symmetry) was assumed, but the framework geometry was strained. An independent evaluation of the symmetry using the powder charge flipping algorithm in Superflip led to a successful refinement with reasonable geometry and a refined composition of |[(C6H3)(C7H10N2)3]2F2|[Si(40.2)Ge(15.8)O112] in the space group Pban.

Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae in a university hospital, Ankara, Turkey.

Penicillin-resistant Streptococcus pneumoniae isolates (n = 76) from clinical samples of patients admitted to Hacettepe University Hospital between January 1997 and December 2001 were included in the study. MICs of penicillin G, erythromycin A, clindamycin, cefaclor, cefotaxime, vancomycin, chloramphenicol, tetracycline, ciprofloxacin and rifampicin were determined by agar dilution. The isolates were serogrouped on the basis of the Neufeld Quellung reaction and were typed by BOX-PCR. Genetic polymorphism of the penicillin resistance genes pbp2b and pbp2x was investigated by restriction fragment length polymorphism (RFLP) analysis. Of the 76 isolates tested, 64 (84.2%) showed intermediate resistance to penicillin, while 12 (15.8%) were resistant to higher levels of penicillin (MIC > or = 2 mg/L). The resistance patterns of the isolates revealed six different resistance profiles. There were 22 different serotypes, with c. 55% of the isolates belonging to serotypes 23B, 19A, 19F, 14, 6 A and 9V. Five distinct patterns for pbp2b and 12 distinct patterns for pbp2x were obtained by RFLP analysis of penicillin-binding protein genes. The combination of these patterns allowed isolates to be classified into 22 fingerprint subgroups. BOX-PCR analysis showed that the isolates fell into 14 distinct BOX genotypes, with 33 subtypes. Serotype 9V isolates with pbp genotype 2-6 and BOX-PCR type 4, 4.1 or 4.2 were related to the pandemic clone Spain(9V)-3. No relatedness to other international clones was detected among the other study strains, but genetic relatedness was observed among some of the serotype 19A and 23B isolates. Overall, the results demonstrated that most of the penicillin-resistant pneumococcal isolates in Turkey, other than those belonging to serotypes 9V, 19A and 23B, were derived from several independent clones, possibly resulting from multiple importation of strains originating from outside the country. Differences in pbp patterns, serotypes and resistance profiles among isolates that showed similar BOX-PCR patterns supported the hypothesis that horizontal transfer of capsular genes, pbp genes and other genetic determinants between S. pneumoniae and viridans group streptococci may have occurred.

Validity of unbound digoxin measurements by immunoassays in presence of antidote (Digibind).

Measurement of unbound digoxin in presence of Fab fragments may be useful in management of overdoses. The analysis can be performed on serum directly or on ultrafiltrate of serum. The architecture of the immunoassay may influence the validity of results obtained using these two approaches. We tested this hypothesis by preparing serum mixtures containing various concentrations of digoxin and Digibind and analyzed them by the immunoassays before and after ultrafiltration. Four samples collected from Digibind-treated patients were also analyzed before and after ultrafiltration. The slopes and the y-intercepts of the measured versus the expected values for serum and its ultrafiltrate overlapped for the MEIA digoxin assay. For other three immunoassays tested (ACS:180, Stratus, and On-Line), either the slope or the intercept for measured versus the expected results for serum were significantly different (P < 0.05) than those for ultrafiltrate. Following addition of digoxin and Digibind, differences in results for serum analyzed directly or after ultrafiltration were < 0.50 ng/ml. Comparable samples from digoxin-overdosed patients treated with Digibind had differences of > 1.0 ng/ml. Previous claims reporting direct analysis of digoxin in presence of antidote but not having used patient samples for validation should be revisited. To date, analysis of serum ultrafiltrate by an immunoassay proven not to have matrix bias remains the most accurate approach in measuring unbound digoxin in presence of antidote.