Brain abscess caused by Nocardia cyriacigeorgica in two patients with multiple myeloma: novel agents, new spectrum of infections.

OBJECTIVE AND IMPORTANCE: Introduction of high-dose chemotherapy and the novel agents including bortezomib, Lenalidomide, and Thalidomide has provided a significant progress in the treatment of multiple myeloma (MM) with an increase in median overall survival up to 6-8 years. However, the advances in myeloma treatment comes at a price with new spectrum of treatment-related infectious complications which should be taken into consideration while treating these patients. CLINICAL PRESENTATION: We report here two patients with Ig G λ MM presenting with intracerebral mass lesions in the abscence of constitutional symptoms that would suggest an infectious etiology. Both patients had severe hypogammaglobulinemia and lymphopenia, which was attributed to treatment regimens including bortezomib. Intervention The surgical intervention-revealed abscess in both cases caused by Nocardia cyriacigeorgica, a relatively new pathogen which rarely causes infections in humans and also an unexpected pathogen in myeloma patients. CONCLUSION: Although every aspect of immune system is known to be affected in MM, humoral immune deficiency is the hallmark of the inherent immune defect in this disease. Introduction of the novel agents, bortezomib in particular seems to have changed the characteristics of the immune dysfunction and the spectrum of the opportunistic infections by causing qualitative and quantitative changes in cellular immunity. The new spectrum of infectious agents might not be limited to hepatitis B and herpes zoster. Monitoring lymphopenia and administration of prophylactic antimicrobial agents accordingly could be considered in patients treated with bortezomib.

The role of interleukin-1 gene polymorphisms and Helicobacter pylori in gastroesophageal reflux disease.

BACKGROUND/AIMS: Our aim is to assess the relationship between interleukin 1ß (IL-1 ß), (-511,-31 alleles), interleukin 1RN (IL-RN), Helicobacter pylori (HP) status and gastroesophageal reflux disease (GERD) diagnosed by pH monitoring in the Turkish population. MATERIALS AND METHODS: A Total of 100 consecutive patients with GERD were enrolled in the study. Genotypes of IL-1ß (-511,-31), IL-1RN gene polymorphisms and HP status of the patients were analyzed. RESULTS: While thirty-two patients were diagnosed as esophagitis with varying severity the remaining patients had no esophagitis. Seventy six participants were positive for HP and the remaining patients were negative. The difference between erosive and non-erosive groups was statistically significant when we compared IL-1ß (-511) but no difference regarding IL-1ß (-31) and IL-1RN variations. We also analyzed T/T, C/T and C/C alleles and the difference was significant statistically in T/T allele between patients with and without erosive GERD 1 (3.1%) vs. 12 (17.9%), respectively with a p value<0.05. But C/C, C/T alleles of (-511), (-31) and IL-1RN polymorphisms were not statistically significant between the groups. CONCLUSION: IL-1ß genetic polymorphisms may take part in the pathophysiology of gastroesophageal reflux disease.

Differential identification of atypical pneumonia pathogens in aorta and internal mammary artery related to ankle brachial index and walking distance.

BACKGROUND: We studied the existence of agents in aorta biopsies, such as Chlamydia pneumoniae, cytomegalovirus, and Mycoplasma pneumoniae, that are thought to have a role in atherosclerosis etiopathogenesis role, and their association with peripheral artery disease. MATERIALS AND METHODS: We examined aorta wall and internal mammarian artery (IMA) biopsies taken from two different places in 63 patients in whom coronary artery bypass was performed. In these biopsies, we evaluated the deoxyribonuclease (DNA) of these microorganisms using polymerase chain reaction. From the same patients, we recorded the ankle brachial index, road walking distance information, lipid profile, C-reactive proteins, blood parameters such as fibrinogen, and the patient's operation data. RESULTS: In the nine aorta biopsies taken from 63 patients, we isolated C pneumoniae DNA. In IMA biopsies taken from the same patients, we detected no microorganism DNA (P < 0.001). In the same aorta biopsies, we found no cytomegalovirus or M pneumoniae DNA. We examined 12 patients using an index value of 0.9 in the ankle brachial index evaluation; eight had C pneumoniae in the aorta biopsies (P < 0.001). CONCLUSIONS: We found a significant relationship between C pneumoniae DNA and the existence of peripheral artery disease. In the development of atherosclerosis with C pneumoniae, there may be a determinant pathogen in both the aorta and the peripheral arteries. The nonexistence of C pneumoniae DNA in the IMA biopsies may indicate infectious agents because of the predominant endothelial functions in this artery, and thus its resistance to atherosclerosis.

[Determination of hepatitis B virus genotypes by DNA sequence analysis in patients from Ankara, Turkey]. / Ankara bölgesindeki hepatit B virusu genotiplerinin DNA dizi analizi yöntemi ile belirlenmesi.

Hepatitis B virus (HBV) genotypes vary depending on the geographical region. The HBV genotype determined in Turkey has been genotype D which is found as the homogenously disseminated single genotype. The aim of this study was to determine HBV genotypes in a group of HBV infected patients who were admitted to a university hospital in Ankara, Turkey. Serum samples from HBsAg positive and anti-HBs negative 84 (52 male, 32 female) patients with HBV infection were included into the study. Anti-HBc was positive in 95.2%, HBeAg was positive in 47.6% and anti-HBe was positive in 11.9% of the patients. Mean HBV-DNA levels of the patients were 5.7 x 10(7) +/- 4.6 x 10(7) IU/ml; mean ALT levels were 131 +/- 171 IU/ml and mean AST levels were 98 +/- 170 IU/ml. HBV-DNA was extracted from serum by the phenol-chloroform method and PCR was performed to amplify the S gene region of HBV-DNA. Cycle sequencing of PCR products was performed by a commercial "Cy5/Cy5.5 Dye Primer Cycle Sequencing Kit" (Visible Genetics, Canada) based on dideoxy chain termination method. The sequences were read and analyzed in an automated fluorescence-based DNA-sequencing system (Long-Read Tower System, Visible Genetics, Canada). The nucleotide sequences of the patient samples were compared with the previously reported sequences in gene bank for each genotype. According to the comparative analysis of S-sequences of all patient samples with the published sequences of the genotypes in gene bank, all of the 84 hepatitis B strains (100%) were shown to be related to D genotypic group, subtype ayw. A phylogenetic analysis was performed and phylogenetic trees were constructed using programs in the PHYLIP phylogeny inference package. The patient samples clustered within the genotypic group D. According to these results, the main HBV genotype in our patients was genotype D in accordance with the previous molecular epidemiologic information on HBV in this geographic area. HBV genotype determination may help to establish more rational clinical approach in the evaluation of HBV infected patients.

In vitro synergistic activity of tigecycline and colistin against XDR-Acinetobacter baumannii.

The emergence of extensive drug-resistant (XDR) Acinetobacter baumannii limits the therapeutic options and leads to high mortality in intensive care units. Combined antibiotic therapy is frequently recommended for the treatment of these infections. Colistin (CO) and tigecycline (TIG), alone or in combination with other antimicrobials, are the most commonly used antibiotics in the treatment of these resistant infections. In this study, the in vitro synergistic activity of TIG and CO were tested for 25 XDR-A. baumannii strains isolated from ventilator-associated pneumonia by the Etest method. Resistance to CO was not detected, whereas 8% of the strains were resistant to TIG. The TIG-CO combination was more synergistic than TIG-rifampin and CO-rifampin according to the fractional inhibitory concentration index. No antagonism was detected between the drugs in the study. There was no strong correlation between the activity of the combinations with reference to strains or genotypes. Our results suggest that the combined use of TIG and CO may be useful for the treatment of XDR-A. baumannii infections.

Topical N-acetylcysteine reduces interleukin-1-alpha in tear fluid after laser subepithelial keratectomy.

PURPOSE: To evaluate the effect of topical N-acetylcysteine (NAC) on interleukin 1-alpha (IL-1alpha) levels in tear fluid after myopic laser subepithelial keratectomy (LASEK) and its possible role in modulating corneal wound healing. METHODS: Twenty-six eyes of 13 patients who underwent myopic LASEK were divided into 2 groups. Group 1 (n=10 eyes) was used as a control group. All patients received topical lomefloxacin and dexamethasone postoperatively. Additionally, patients in Group 2 received topical NAC for 1 month postoperatively. Tear fluid samples were collected with microcapillary tubes preoperatively, on the first and on the fifth postoperative day, and the release of IL-1alpha in tear fluid was calculated. Haze grading and confocal microscopic examination were performed at 1 and 3 months postoperatively. RESULTS: The mean IL-1-alpha release values were 0.285-/+0.159 pg/min in Group 1 and 0.235-/+0.142 pg/min in Group 2 preoperatively. In Group 1, the values were 0.243-/+0.155 pg/min on day 1 and 0.164-/+0.125 pg/min on day 5. In Group 2, the mean IL-1alpha release values were 0.220-/+0.200 pg/min on day 1 and 0.080-/+0.079 pg/min on day 5. The difference between the groups was significant only for day 5 (p<0.05). Mean corneal haze score and grey scale value in confocal microscopy were significantly higher (p<0.05) in Group 1 at 1 month. However, at 3 months there was no difference between groups (p>0.05). CONCLUSIONS: NAC seems to have an additive effect to steroids in suppressing IL-1alpha levels in tear fluid and may be clinically advantageous in modulating corneal wound healing during the early postoperative period after LASEK.

Detection of Helicobacter pylori DNA in aortic and left internal mammary artery biopsies.

We investigated the relationship between acute coronary ischemia and the presence of Helicobacter pylori DNA in aortic regions that were absent macroscopic atheromatous plaques. The study group (Group 1) consisted of 42 patients who underwent coronary artery bypass grafting. Biopsy samples were obtained from 2 different locations: from regions of the aorta that were free (macroscopically) of atheromatous plaque (Group 1A), and from the internal mammary artery (Group 1B). The control group (Group 2) of 10 patients who had no atherosclerotic vascular disease provided aortic tissue samples for comparison. The real-time polymerase chain reaction method was used to detect H. pylori DNA in all biopsy samples. Eleven of 42 aortic tissue samples (26%) in Group 1A were positive for H. pylori DNA. Neither biopsies from the left internal mammary arteries of those patients nor biopsies from the aortas of the control group (Group 2) were positive for H. pylori DNA. There was a statistically significant difference between 1A and 1B in terms of H. pylori positivity (P=0.001). In Group 1 as a whole, acute coronary ischemia was more prevalent in the H. pylori-positive patients than in the H. pylori-negative patients (P=0.001). To our knowledge, this is the 1st study to investigate the detection of H. pylori DNA in aortic biopsy samples that are macroscopically free of atheromatous plaque. Such detection in patients who have atherosclerotic coronary artery disease could be an important indication of the role of microorganisms in the pathogenesis of atherosclerosis.

Polymorphisms in the aurora-A gene is not associated with lung cancer in the Turkish population.

Lung cancer, a complex neoplasm of lung tissue, is influenced by several environmental and genetic factors which could be changed in each individual. Aurora-A gene is related to mitotic events such as: chromosome instability, cell cycle regulation, spindle formation, and kinetechore-microtubule connections. This centrosomic serine/threonine kinase provides a strong connection between mitotic errors and carcinogenesis. The genomic alterations such as single nucleotide polymorphisms (SNPs) can exist in molecular pathways of lung cancer. Therefore, we evaluated the role of genetic polymorphisms of Aurora-A gene in the lung cancer in the Turkish population. Genotypes of five Aurora-A polymorphisms (F31I, V57I, 6328G/A, P50L, and S104L) were determined in 102 healty controls and 102 new diagnosed lung cancer cases. All samples were genotyped with DNA sequence technique. There were not any genotype variations in P50L, S104L, and 6328G/A polymorphisms. The frequencies of both genotypes F31I and V57I in lung cancer patients were not significantly different from those in controls (p > 0.05). A multivariable logistic regression analysis including patient characteristics, such as age and gender, did not change the results.

Microbiologic study of soft contact lenses after laser subepithelial keratectomy for myopia.

PURPOSE: To evaluate the extent and agents of bacterial contamination of bandage disposable soft contact lenses after laser subepithelial keratectomy (LASEK) and to correlate the findings with clinical data. METHODS: Disposable soft contact lenses were collected from 52 eyes of 26 consecutive patients treated with LASEK for myopia. The patients were treated with a fixed combination of tobramycin and diclofenac until epithelial closure. The lenses were removed on the fourth or fifth postoperative day with sterile forceps and immediately placed in sterile tubes containing culture media brain-heart infusion broth. The lenses were evaluated for microbial colonization. RESULTS: Of the 52 contact lenses analyzed, six (11.5%) had positive cultures. However, no clinical finding of infection was noted. Isolated microorganisms were coagulase-negative staphylococci (two lenses), Stenotrophomonas maltophilia (two lenses), Acinetobacter species (one lens), and Aeromonas hydrophila (one lens). Except for one case, the microorganisms were sensitive to the administered antibiotic. CONCLUSIONS: The risk of infectious keratitis after LASEK seems to be low. Except for staphylococci, the isolated microorganisms have not been previously reported to colonize the ocular surface or cause keratitis after refractive surgery. These findings may suggest a changing trend of potentially infectious agents after surface ablation.

Identification of Helicobacter species by 16S rDNA PCR and sequence analysis in human liver samples from patients with various etiologies of benign liver diseases.

BACKGROUND/AIMS: Several reports indicated an increased prevalence of the Helicobacter species in hepatocellular cancer tissue and in liver samples infected with hepatitis viruses. The frequency of Helicobacter spp. in benign liver diseases was, however, not thoroughly investigated. METHODS: Seventy-five consecutive patients with suspected liver disease were enrolled. The indications were hepatitis B virus (n=30), C virus (n=8), B and C dual infection (n=1), nonalcoholic steatohepatitis (n=27), autoimmune hepatitis (n=3), primary biliary cirrhosis (n=1) and idiopathic elevation of liver enzymes (n=5). PCR detection of 16S recombinant RNA gene of Helicobacter spp. was performed on liver samples. PCR products of positive samples were further identified by DNA sequencing. The patients also had upper gastrointestinal endoscopy and gastric biopsy for the detection of H. pylori using histopathology and PCR. RESULTS: Helicobacter spp. DNA was detected in two out of 75 liver biopsy samples (2.6%), which were typed as H. pylori by DNA sequencing. One of these patients had chronic hepatitis C infection (man, 51 years old) and the other had nonalcoholic steatohepatitis (woman, 44 years old). Fifty-two out of 75 of the patients (69.3%) had H. pylori infection in their stomachs. CONCLUSION: We have found that H. pylori infection is much less prevalent in benign liver diseases. The presence of H. pylori in nonalcoholic steatohepatitis (NASH) patients is a novel finding and this finding should be confirmed in a larger series.

Diagnosis of Helicobacter pylori.

Although there are attempts to perform Helicobacter pylori diagnosis directly in vivo using magnification endoscopy, most articles on diagnosis this year concerned non-invasive tests and molecular methods. For urea breath tests, there are attempts to have a quicker and cheaper test and to evaluate its role in cases of premalignant lesions. For stool antigens tests, evaluation of kits using monoclonal antibodies was carried out. Molecular tests have been applied for typing and detection of resistant mutants.

Rubella seroprevalence among women of childbearing age residing in a rural region: is there a need for rubella vaccination in Turkey?

It is essential to evaluate the susceptibility of women in the reproductive age group to rubella virus in order to set strategies for the prevention of congenital rubella syndrome (CRS). Turkey began implementing measles-mumps-rubella vaccination as part of the national vaccination schedule for children (12 months, 6 years) and adolescents (14 years) in July, 2006, and there is an ongoing discussion of the need for a policy of vaccinating women of child-bearing age against rubella. The aim of this study was to determine the rubella seroprevalence among women in the reproductive age group in a rural district in Ankara and to provide data about rubella susceptibility for policymakers. Four hundred ninety of the women in the 15- to 49-year-old age group in the region who were targeted were reached (68.2%), and 467 (65.0%) of them who had a convenient serology were included in the study. Rubella IgG antibodies were quantified by the enzyme-linked immunosorbent assay method. Seropositivity was 95.5% for the total group and 96.2% among pregnant women. The seropositivity of this rural group of women was found to be high, but in order to rule out the need for a rubella vaccination program for women of child-bearing age, large-scale studies in different settings and studies that describe the CRS burden in Turkey are required.

Effects of atypical pneumonia agents on progression of atherosclerosis and acute coronary syndrome.

BACKGROUND: The aim of this study was to investigate the presence of various atypical pneumonia agents (Chlamydia pneumoniae, cytomegalovirus, Mycoplasma pneumoniae), which are considered to have a role in the ethiopathogenesis of atherosclerosis, in aortic biopsies without macroscopically visible plaque and in internal thoracic artery biopsies. MATERIAL AND METHODS: Thirty-three patients (group 1), who had undergone coronary bypass operation and 10 non-atherosclerotic patients (group 2), were included in the study. Seventy-six tissue biopsies were taken. Biopsies from the patients in group 1 a were obtained from the atheroma plaque-free aortic tissue and 33 biopsies (group Ib) were obtained from their internal thoracic arteries. Following DNA extraction, nested PCR was used to detect Chlamydia pneumoniae DNA, and real time PCR was used to detect cytomegalovirus and Mycoplasma pneumoniae DNA. Blood parameters (lipid profile, CRP, fibrinogen) of the patients and operation characteristics were recorded. RESULTS: Chlamydia pneumoniae DNA was detected in 5 of 33 biopsy samples from coronary bypass patients, whereas none of the control patients (group 1b and group 2) were positive for this agent (P = 0.001). Neither CMV nor Mycoplasma pneumoniae was detected in IMA and aortic biopsies of both bypass and control patients. Elevated total cholesterol levels (P = 0.02) and positive CRP (P = 0.001) was found in C. pneumoniae positive patients. Prevalence of acute coronary syndrome was significantly higher in C. pneumoniae detected patients compared (P = 0.00 1). CONCLUSIONS: Detection of C. pneumoniae DNA in the atheroma free aortic biopsies might indicate that this micro-organism intervened in the progression of atheroma plaque. There was a strong relationship between the detection of this micro-organism in the aortic wall and acute coronary syndrome. The absence of DNA of the corresponding micro-organisms in the IMA wall may show its resistance to infective agents and in turn to atherosclerosis, which is a result of the prevailing endothelial functions of this artery.

A follow up study of cytomegalovirus infection in a group of Turkish renal transplant recipients using molecular assays.

The clinical value of an in-house cytomegalovirus nested polymerase chain reaction (CMV-PCR) and a commercial molecular assay hybrid capture CMV DNA assay (HCA) was evaluated in monitoring a group of renal transplant patients for six months follow up. In this study, the sensitivity, specificity, positive predictive value, and negative predictive value of nested CMV DNA PCR assay and HCA at the beginning of the study were 70, 42.9, 46.7, 66.7, and 60, 78.6, 66.7, and 73.3% respectively. After six months, they were 80, 66.7, 80, 66.7 for CMV PCR and 73.3, 88.9, 91.7, 66.7% for HCA respectively. These results indicate that in monitoring and predicting CMV infections in renal transplant recipients, not only qualitative but also quantitative assays must be used together in order to decide the preemptive strategies.

A follow up study of cytomegalovirus infection in a group of Turkish renal transplant recipients using molecular assays

The clinical value of an in-house cytomegalovirus nested polymerase chain reaction (CMV-PCR) and a commercial molecular assay hybrid capture CMV DNA assay (HCA) was evaluated in monitoring a group of renal transplant patients for six months follow up. In this study, the sensitivity, specificity, positive predictive value, and negative predictive value of nested CMV DNA PCR assay and HCA at the beginning of the study were 70, 42.9, 46.7, 66.7, and 60, 78.6, 66.7, and 73.3 percent respectively. After six months, they were 80, 66.7, 80, 66.7 for CMV PCR and 73.3, 88.9, 91.7, 66.7 percent for HCA respectively. These results indicate that in monitoring and predicting CMV infections in renal transplant recipients, not only qualitative but also quantitative assays must be used together in order to decide the preemptive strategies.

Use of molecular methods in identification of Candida Species and evaluation of fluconazole resistance

The aim of this study was to evaluate the use of one of the molecular typing methods such as PCR (polymerase chain reaction) following by RFLP (restriction fragment length polymorphism) analysis in the identification of Candida species and then to differentiate the identified azole susceptible and resistant Candida albicans strains by using AP-PCR (arbitrarily primed-polymerase chain reaction). The identification of Candida species by PCR and RFLP analysis was based on the size and primary structural variation of rDNA intergenic spacer regions (ITS). Forty-four clinical Candida isolates comprising 5 species were included to the study. The amplification products were digested individually with 3 different restriction enzymes: HaeIII, DdeI, and BfaI. All the isolates tested yielded the expected band patterns by PCR and RFLP analysis. The results obtained from this study demonstrate that Candida species can be differentiated as C. albicans and non-C. albicans strains only by using HaeIII restriction enzyme and BfaI maintains the differentiation of these non-C. albicans species. After identification Candida species with RFLP analysis, C. albicans strains were included to the AP-PCR test. By using AP-PCR, fluconazole susceptible and resistant strains were differentiated. Nine fluconazole susceptible and 24 fluconazole resistant C. albicans were included to the study. Fluconazole resistant strains had more bands when evaluating with the agarose gel electrophoresis but there were no specific discriminatory band patterns to warrant the differentiation of the resistance. The identification of Candida species with the amplification of intergenic spacer region and RFLP analysis is a practical, short, and a reliable method when comparing to the conventional time-consuming Candida species identification methods. The fluconazole susceptibility testing with AP-PCR seems to be a promising method but further studies must be performed for more specific results.

Detection of Helicobacter pylori and its CagA gene in tonsil and adenoid tissues by PCR.

OBJECTIVE: To determine the presence of Helicobacter pylori and, if detected, the prevalence of the CagA gene in adenotonsillectomy specimens by polymerase chain reaction (PCR). DESIGN: A prospective clinical trial. SETTING: Tertiary referral center. PATIENTS AND METHODS: The study population comprised 23 patients who had undergone adenoidectomy, tonsillectomy, or adenotonsillectomy under local or general anesthesia. Helicobacter pylori DNA was extracted from 3-mm-diameter tissue samples obtained from each tonsil and adenoid tissue specimens. The amplifications were performed for the 16S ribosomal RNA (rRNA) and CagA genes of H pylori in the samples of which H pylori DNA was detected. RESULTS: In examining all the samples, 7 (30%) of 23 patients were shown to be positive for H pylori DNA, 5 (71%) of whom also possessed the CagA gene. CONCLUSIONS: Tonsil and adenoid tissues may be an ecological niche of the mouth without regard to transient or permanent colonization. Oral-oral transmission may be a possible mode of spread of H pylori.

Effects of lactulose and lactitol on coliform bacteria and bacterial translocation in the caecum during 72-h starvation in rats.

Lactulose and lactitol, non-absorbable disaccharides, prevent bacterial translocation (BT) arising from the gut. In contrast, lack of food into the gut leads to coliform bacterial overgrowth and even if it does not cause BT, can induce the risk from other stimuli for BT. In this study, we tested whether lactulose and lactitol affected populations of coliform bacteria in the caecum during starvation in Sprague-Dawley rats. Three groups of rats were starved for 72 h and given oral 2 ml undiluted lactulose (670 mg/ml), 2 ml undiluted lactitol (666 mg/ml) or 2 ml physiological saline, respectively, once a day. The caecum and mesenteric lymph nodes (MLNs) were removed for microbiological and histopathological analyses. The highest degree of coliform bacterial overgrowth, BT to MLNs and histopathological damage were observed in lactulose-treated rats, followed by the group treated with lactitol. As a result of this study, both drugs, especially lactulose augmented the proliferation and translocation tendency of coliform bacteria in the caecum during 72-h starvation in rats.

A possible role of Helicobacter pylori in chronic rhinosinusitis: a preliminary report.

OBJECTIVE: The objective was to investigate the presence of Helicobacter pylori with polymerase chain reaction in the sinonasal mucosa of patients with or without chronic rhinosinusitis (CRS). STUDY DESIGN: A prospective clinical trial. METHODS: Mucosal tissue samples were collected from ethmoid cells of 12 patients with CRS and the removed mucosal part of the middle concha of 13 patients with concha bullosa who were treated surgically in our institution. DNA extracted from these samples was used for the amplification of 16S ribosomal RNA gene of H pylori by nested polymerase chain reaction. RESULTS: Helicobacter pylori DNA was detected in 4 of 12 patients with CRS, but it was not detected in patients with concha bullosa. Three of four patients with positive results for H pylori had gastroesophageal reflux-related complaints. CONCLUSIONS: It is possible to detect H pylori in the sinus mucosa of some patients with CRS. However, whether H pylori is a causative agent for CRS or a result of CRS is not known.