Comparison of Methods in the Serologic Diagnosis of Cystic Echinococcosis.

PURPOSE: Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests. METHODS: In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard. RESULTS: Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ2 = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test. CONCLUSION: There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability.

Comparison of homologous and heterologous inactivated and mRNA vaccination programme against SARS-CoV-2 in dialysis patients.

AIM: To evaluate the vaccine response and the effect of the booster dose on COVID-19 positivity in haemodialysis (HD) and peritoneal dialysis (PD) patients who received and did not receive BNT162b2 as a booster dose after two doses of CoronaVac. METHODS: The study included 80 PD and 163 HD patients, who had been administered two doses of the CoronaVac. Antibody levels were measured on Days 42 and 90 after the first dose. Measurements were repeated on Day 181 after the first dose in the patients that received two vaccine doses and on Day 28 after the third dose in those that also received the booster dose. Antibody levels below 50 AU/mL were considered negative. RESULTS: The seropositivity rate was similar in the HD and PD group on Days 42 and 90 (p = 0.212 and 0.720). All patients were seropositive in the booster group. The antibody level was lower in the patients that received CoronaVac as the booster compared to those administered BNT162b2 in HD and PD groups (p < 0.001 and 0.002). COVID-19 positivity was detected in 11 patients (7 = had not received the booster dose, 4 = had received third dose of CoronaVac). The multivariate analysis revealed that as age increased, COVID-19 positivity also increased (OR: 1.080, 95% CI: 1.017 - 1.146, p = 0.012), while booster dose administration decreased this positivity (OR: 0.113, 95% CI: 0.028 - 0.457, p = 0.002). CONCLUSION: Our results may indicate the need for additional vaccination doses in patients with HD and PD. Our findings indicate a higher antibody response in dialysis patients with heterologous BNT162b2 as a booster dose after two doses of CoronaVac compared to homologous CoronaVac.

Assessing neutrophil-derived ROS production at the bedside: a potential prognostic tool in severe COVID-19 cases.

PURPOSE: A prompt and effective immune response is required for clearance of pathogens but exaggerated states of inflammation can cause extensive collateral damage to the host. We have previously used a rapid near-patient assay that measures the functional capacity of neutrophils to produce reactive oxygen species (ROS) to show that values are elevated in patients with severe COVID-19 or sepsis. Here, we assess the utility of longitudinal ROS measurements to monitor and predict mortality outcome for patients with COVID-19 infection being treated in an ICU setting. METHODS: We used the Leukocyte ImmunoTest™ (LIT™) to quantify neutrophil ROS release using a small volume (10 µL) of capillary blood in a portable, rapid (10-min) format. RESULTS: ROS values (LIT score) and ROS levels assessed in relation to neutrophil count (LIT/N) were both markedly elevated in the patient group. Furthermore, these correlated strongly with peripheral neutrophil count and CRP value. Serial measurement of neutrophil or CRP values were not able to reliably predict mortality within the study. In contrast, LIT and LIT/N values started to decline at 7 and 5 days, respectively, in patients who survived ICU admission and this increment increased further thereafter. CONCLUSIONS: This study raises the possibility of LIT and LIT/N to be used as a predictive clinical tool for patients with severe COVID-19 and argues for its assessment to inform on prognosis, and potentially guide treatment pathways, in other disorders associated with neutrophil activation. TAKE-HOME MESSAGE: A longitudinal study of 44 severe COVID-19 patients in the ICU of a leading teaching hospital has demonstrated the prognostic potential of a rapid bedside assay of neutrophil-derived reactive oxygen species (ROS). Assessment of changes in ROS production, as measured using the Leukocyte ImmunoTest™, shows that ROS production generally declined back to normal levels for patients who survived, but remained elevated for those patients who did not survive.

Establishment of a method to detect human immunoglobulin G1 binding microorganisms.

In this study, we developed an original and rapid anti-idiotypic antibody-based ELISA method, different from the techniques defined in the literature for this purpose so far, to detect immunoglobulin binding proteins (IBP) on the surface of bacteria. The test antibody used in our study to detect IBP is a recombinant human immunoglobulin G1 Kappa molecule, and has been used as a drug, Tocilizumab (Actemra®), in humans for therapeutic purposes. As a result, the test antibody in the supernatant after centrifugation is reduced compared to the initial moment due to antibody binding. Staphylococcus aureus cowan 1 strain used as positive control causes at least a 50% decrease in OD value in this respect. A similar observation at this level indicated that among a total of 189 microorganisms tested, 3 Staphylococcus aureus and 1 MRSA carrying high-affinity IgBP showed greater than 50% inhibition. This level of inhibition was not detected in the remaining microorganisms.

[Investigation of Group A Rotavirus G10, G12 Genotypes Emerging in Patients with Acute Gastroenteritis in a Tertiary Care Hospital]. / Hastanemize Basvuran Akut Gastroenteritli Hastalarda Yeni Görülmeye Baslayan A Grubu Rotavirüs G10, G12 Genotiplerinin Arastirilmasi.

Rotaviruses are the most common cause of viral gastroenteritis with the highest mortality and morbidity rates in children aged 0-5 years. The aim of this study was to determine the frequency of rotavirus infection in patients whose stool samples were sent to microbiology laboratory to investigate the etiology of diarrhea, to investigate the rotavirus genotypes that are common in our region and G10, G12 genotypes that have recently become common in the world. Fecal samples of 476 patients aged between 0-92 years who applied between November 2016 and February 2018 were studied via immunochromatographic rapid test and enzyme-linked immunosorbent assay (ELISA) methods. ELISA positive samples were studied by nested reverse transcriptase chain reaction (RT-PCR) and genotyped by agarose gel electrophoresis. Rotavirus was found positive in 18.3% and 17% of stool samples by immunochromatographic test and ELISA, respectively. All ELISA positive samples were also detected as positive by RT-PCR. 18.5% of female patients and 15.7% of male patients were found to be positive and rotavirus positivity was not statistically significant between genders. The frequency of rotavirus in different age groups was 23.5% (6-12 years), 17.3% (13-24 months) and 16% (25-36 months). It was determined that rotavirus cases were most common in the spring. G1, G2, G3, G4, G9, G10, and G12 were detected in 37%, 7.4%, 16.1%, 6.2%, 9.9%, 2.5%, 26% of the samples, respectively. G12 was the most common genotype after G1. The most common G and P genotype combination was G1P[8] (17.2%). This was followed by G12P[8] (11.11%) and G3P[8] (11.11%). P[8] (53%) was found to be the dominant P genotype. In this study, it was observed that rotavirus, which is the cause of childhood diarrhea, can also be encountered in advanced ages and even new genotypes that infect humans worldwide may also be the causative agents. Therefore, we concluded that it is important to investigate new genotypes such as G10 and G12 in molecular epidemiological studies.

Case Report: A Human Case of Onchocerca lupi Mimicking Nodular Scleritis.

We report a case of human ocular onchocerciasis by zoonotic Onchocerca lupi presenting as nodular scleritis. Molecular analyses were used to confirm diagnosis at species level. In addition to few existing reports of human infection by O. lupi in Turkey, this case further suggests that the pathogen might be more common than previously reported, thus requiring further attention and investigations.

Genomic characterization of SARS-CoV-2 isolates from patients in Turkey reveals the presence of novel mutations in spike and nsp12 proteins.

Novel mutations have been emerging in the genome of severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2); consequently, the evolving of more virulent and treatment resistance strains have the potential to increase transmissibility and mortality rates. The characterization of full-length SARS-CoV-2 genomes is critical for understanding the origin and transmission pathways of the virus, as well as identifying mutations that affect the transmissibility and pathogenicity of the virus. We present an analysis of the mutation pattern and clade distribution of full-length SARS-CoV-2 genome sequences obtained from specimens tested at Gazi University Medical Virology Laboratory. Viral RNA was extracted from nasopharyngeal specimens. Next-generation sequencing libraries were prepared and sequenced on Illumina iSeq 100 platform. Raw sequencing data were processed to obtain full-length genome sequences and variant calling was performed to analyze amino acid changes. Clade distribution was determined to understand the phylogenetic background in relation to global data. A total of 293 distinct mutations were identified, of which 152 missense, 124 synonymous, 12 noncoding, and 5 deletions. The most frequent mutations were P323L (nsp12), D614G (ORF2/S), and 2421C>T (5'-untranslated region) found simultaneously in all sequences. Novel mutations were found in nsp12 (V111A, H133R, Y453C, M626K) and ORF2/S (R995G, V1068L). Nine different Pangolin lineages were detected. The most frequently assigned lineage was B.1.1 (17 sequences), followed by B.1 (7 sequences) and B.1.1.36 (3 sequences). Sequence information is essential for revealing genomic diversity. Mutations might have significant functional implications and analysis of these mutations provides valuable information for therapeutic and vaccine development studies. Our findings point to the introduction of the virus into Turkey through various sources and the subsequent spread of several key variants.

The prognostic role of neopterin in COVID-19 patients.

In Coronavirus disease-2019 (COVID-19) cases, hyper inflammation is associated with the severity of the disease. High levels of circulating cytokines were reported in severe COVID-19 patients. Neopterin produced by macrophages on stimulation with interferon-gamma, which is an important cytokine in the antiviral immune response, hence it can be used to predict the severity of disease in COVID-19 cases. In this study, it was aimed to determine the prognostic value of the neopterin for the prediction of severe disease in patients with COVID-19. This single-center, prospective study was conducted in hospitalized COVID-19 patients and healthy volunteers. Severe and mild COVID-19 cases were compared in terms of clinical and laboratory findings as well as serum neopterin levels on hospital admission. To assess the prognostic utility of neopterin between the severe and mild COVID-19 groups, a receiver-operating characteristic (ROC) curve was generated, and the area under the curve (AUC) was calculated. The median serum neopterin level was four times higher in COVID-19 patients than the healthy controls (46 vs. 12 nmol/L; p < .001). The AUC value of serum neopterin was 0.914 (95% confidence interval, 0.85-0.97). The sensitivity and specificity of serum neopterin for the cut-off value of 90 nmol/L to identify severe COVID-19 cases were 100% and 76%, respectively. Serum neopterin levels on hospitalization were significantly higher in severe COVID-19 disease than mild COVID-19 patients. Neopterin levels can be used as an early prognostic biomarker for COVID-19 on admission.

Monoclonal outbreak of Ralstonia solanacearum catheter-related bloodstream infection associated with contaminated package of normal saline solution in a tertiary care hospital

Background/aim: Ralstonia solanacearum is a very rare cause of infection in humans. There is no described nosocomial outbreak due to R. solanacearum so far. We determined R. solanacearum as the source of catheter-related bloodstream infection (CRBSI) outbreak. Materials and methods: This outbreak analysis was carried out in a 1000-bed tertiary care university hospital in Turkey. The outbreak analysis included hematology, oncology, nephrology, gastroenterology wards, emergency department, and intensive care units. The first case with R. solanacearum CRBSI was detected on May 20, 2019 and R. solanacearum was isolated in catheter blood cultures in 34 patients until October 3, 2019 Results: Standard outbreak analysis procedures were applied. Culture samples were taken from the fluids administered via catheters. The cultures did not yield any bacteria. As a result of the investigation in storage area, it was found that there were leaks, air bubbles, and water drops inside the packaging of saline solutions. R. solanacearum was yielded in the cultures obtained from the surface of saline bags and the inner sides of plastic packings. To validate our hypothesis, a clonal analysis was performed using arbitrarily primed-PCR method and Sanger sequencing of the 16S rRNA gene for identification among isolates. All R. solanacearum isolates were monoclonal and identical. Conclusion: This is the first outbreak of R. solanacearum CRBSI described in a hospital setting. The source of the outbreak was a contamination in the surface of saline bags and the inner sides of plastic packings. Efficacy of an active surveillance system, accurate and rapid conduction of microbiological identification are essential for outbreak management.

The factors predicting pneumonia in COVID-19 patients: preliminary results from a university hospital in Turkey

Background/aim: Pneumonia is the most serious clinical presentation of COVID-19. This study aimed to determine the demographic, clinical, and laboratory findings that can properly predict COVID-19 pneumonia. Materials and methods: This study was conducted in the Gazi University hospital. All hospitalized patients with confirmed and suspected SARS-CoV-2 infection between 16 March 2020 and 30 April 2020 were analyzed retrospectively. COVID-19 patients were separated into two groups, pneumonia and nonpneumonia, and then compared to determine predicting factors for COVID-19 pneumonia. Variables that had a P-value of less than 0.20 and were not correlated with each other were included in the logistic regression model. Results: Of the 247 patients included in the study 58% were female, and the median age was 40. COVID-19 was confirmed in 70.9% of these patients. Among the confirmed COVID-19 cases, 21.4% had pneumonia. In the multivariate analysis male sex (P = 0.028), hypertension (P = 0.022), and shortness of breath on hospital admission (P = 0.025) were significant factors predicting COVID-19 pneumonia. Conclusion: Shortness of breath, male sex, and hypertension were significant for predicting COVID-19 pneumonia on admission. Patients with these factors should be evaluated more carefully for diagnostic procedures, such as thorax CT.

[Characterization of Klebsiella pneumoniae strains exposed to subinhibitory concentrations of chlorhexidine]. / Düsük düzeyde Klorhekzidin ile karsilastirilmis Klebsiella pneumoniae izolatlarinin karakterizasyonu.

Chlorhexidine, a topical antiseptic, acts as a cationic biguanide altering the osmotic transport of the bacterial cell wall that has been used throughout the world to prevent healthcare-associated infections for decades. The routine application of chlorhexidine can result in decreased susceptibility of bacteria over time. The aim of this study was to develop Klebsiella pneumoniae strains after exposure to chlorhexidine and characterize these adapted strains in terms of their virulence ability both by in vivo and in vitro methods. Two clinical strains of K.pneumoniae were included in the study. One strain was completely susceptible and the other was resistant to certain antibiotics. Susceptible strain was subjected in the exposure assay as parent/wild strain. Exposure was performed by increasing chlorhexidine concentrations in agar plates. Chlorhexidine concentrations were gradually decreased reaching a final concentration of 0.12 mg/L after five weeks. Chlorhexidine-adapted viable colonies were selected and isolated. Minimal inhibitor concentrations of chlorhexidine, sodium hypochloride, benzalkonium chloride and triclosan for K.pneumoniae strains were determined using broth microdilution method. Reverse transcription-polymerase chain reaction analysis were performed for efflux pumps named cepA, kdeA and acrKp expressions. Fluorimetric efflux assay by using Rhodamine 6G was performed. Galleria mellonella killing assay and in vitro virulence determinants such as esculin hydrolysis, biofilm production, lecithinase, DNase activity, hemolytic activity, lipase production, mucoviscocity, casein hydrolysis and complement-mediated serum killing were evaluated. K.pneumoniae strains exposed to chlorhexidine did not show any antibiotic resistance. MICs for chlorhexidine, sodium hypochloride, and benzalkonium chloride were increased in the adapted strain. Efflux pumps of cepA and kdeA were over-expressed in the chlorhexidine adapted strain. Rhodamine 6G assay showed an increased efflux in the adapted strain. G.mellonella killing assay showed median virulence score. All strains, were esculin positive, while biofilm production, lecithinase, DNase, hemolytic activity, lipase production, mucoviscocity, casein hydrolysis were all negative. The susceptible parent/wild strain was susceptible to the complement-mediated serum killing, while the chlorhexidine adapted strain showed intermediate susceptibility. Chlorhexidine adapted strains of K.pneumoniae showed increased efflux pump expression, enhanced G.mellonella killing and raised resistance to serum killing. No difference was determined for other determinants. Minimal correlation was found between chlorhexidine resistance and virulence in K.pneumoniae.

The in vitro effect of antimicrobial photodynamic therapy on Candida and Staphylococcus biofilms

Background/aim: This study was designed to evaluate the effect of antimicrobial photodynamic treatment (APDT) in a biofilm model using combinations of various dyes (rose bengal, riboflavin, and methylene blue) as photosensitizers and light sources (LED and UVA) against staphylococcal and candidal biofilms. Materials and methods: Sterile microtiter plates were used for the development and quantification of the biofilms. APDT was carried out using combinations of the light sources and dyes. The percentage of the growth inhibition was then calculated using a spectrophotometer. The broth media in the wells were aspirated, wells were stained with crystal violet, and optical density values were measured spectrophotometrically. SEM analysis of the impact of APDT on bacterial and fungal biofilms was also performed. Results: The experiments showed that the most efficacious combination was red LED + methylene blue against both staphylococcal and candidal biofilms. A marked inhibition (45.4%) was detected on both C. albicans and C. parapsilosis biofilms. Red LED + methylene blue was also effective on S. aureus and S. epidermidis biofilms. SEM images suggested that the number of adherent cells and biofilm mass were markedly reduced after APDT treatment. Conclusion: Although the results of this study indicated the in vitro efficacy of APDT, it might also be a promising technique for the control of biofilm growth within intravenous catheters.

Successful treatment of refractory listeria meningitis and bacteremia with trimethoprim-sulfamethoxazole in an immunocompetent child.

Listeria monocytogenes is an important cause of life-threatening bacteremia and meningoencephalitis in neonates, pregnant women, the elderly, and immunocompromised individuals. However, it is an uncommon cause of illness in immunocompetent children beyond the neonatal period. Ampicillin with or without an aminoglycoside remains the best treatment for listeriosis. Here, we report a rare case of Listeria meningitis and bacteremia in a 7-month-old immunocompetent girl, which was refractory to ampicillin plus gentamicin treatment and successfully treated by the addition of TMP/SMX.

Microbiologic Examination of Bandage Contact Lenses Used after Corneal Collagen Cross-linking Treatment.

PURPOSE: To investigate the agents of bacterial contamination of contact lenses after corneal collagen cross-linking (CCL), and to present the possible changes of ocular flora after riboflavin/ultraviolet A. METHODS: Seventy-two contact lenses of patients who underwent CCL and 41 contact lenses of patients who underwent photorefractive keratectomy (PRK) as control group were enrolled to the study. After 48 h of incubation, broth culture media was transferred to plates. Samples were accepted as positive if one or more colony-forming units were shown. RESULTS: There were positive cultures in 12 (16.7%) contact lenses in the CCL group and 5 (12.2%) had positive cultures in PRK group. Coagulase-negative staphlycocci (CNS) were the most frequent microorganism. Alpha hemolytic streptococci and Diphteroid spp. were the other isolated microorganisms. CONCLUSIONS: Bacterial colonization can occur during and early after the CCL procedure in epithelial healing. To prevent corneal infections after the treatment, prophylactic antibiotics should be prescribed.

[Using Galleria mellonella as an in vivo model to study the virulence of some bacterial and fungal agents]. / Bazi bakteri ve mantarlarin virülansinin arastirilmasinda Galleria mellonella'nin in vivo model olarak kullanilmasi.

Non-vertebrate hosts, such as Galleria mellonella, namely wax moth, have been used to study microbial virulence and host defense. This organism has advantages as it is economical, ethically expedient and easy to handle. Here we describe an experimental in vivo study using the larvae of Galleria mellonella infected with some bacterial and fungal pathogens. In this study, extended-spectrum beta-lactamase (ESBL) producing and non-producing Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, colistin resistant and susceptible Acinetobacter baumanii clinical strains; Candida albicans (ATCC 10231), Scedosporium aurantiacum (CBS 136047) and Pseudallescheria boydii (CBS 117410) reference strains, and Aspergillus terreus and Fusarium oxysporum clinical strains were used as pathogens. The larvae of G.mellonella were challenged with these bacterial and fungal strains, and the mortality rates were calculated using Kaplan-Meier plots. Mortality rates at 16th hour were found as 83% for the larvae infected with both ESBL positive and negative E.coli, ESBL negative K.pneumoniae and ESBL positive P.aeruginosa; 91% for ESBL positive K.pneumoniae; 75% for ESBL negative P.aeruginosa; 66% for both colistin resistant and susceptible A.baumanii strains. All larvae infected with bacteria died within the first 24 hour. Larvae infected with bacteria showed significantly higher mortality rates than those infected with fungi. Mortality rates at 16th hour were found as 0% for C.albicans and F.oxysporum, 16% for S.aurantiacum, 8% for P.boydii and A.terreus; at 24th hour that was 25% for C.albicans and P.boydii, 33% for S.aurantiacum, A.terreus and F.oxysporum; at 48th hour that was 33% for C.albicans, 50% for P.boydii and F.oxysporum, 58% for A.terreus, and 66% for S.aurantiacum; in 72 hours that was 58% for C.albicans and F.oxysporum, 66% for P.boydii, 75% for A.terreus and S.aurantiacum, in 96 hours that was 83% for C.albicans, P.boydii and F.oxysporum, 91% for A.terreus and S.aurantiacum. As a result of this study, potential evidences provided that bacteria were more virulent than fungi for G.mellonella larvae model, each fungal species showed different virulence patterns, and bacterial virulence was correlated neither with species nor antibiotic susceptibility.

[Investigation of the Presence of Disinfectant Resistance Genes qacA/B in Nosocomial Methicillin-Resistant Staphylococcus aureus Isolates and Evaluation of Their In Vitro Disinfectant Susceptibilities]. / Hastane Enfeksiyonu Etkeni Olan Metisiline Dirençli Staphylococcus aureus Suslarinda qacA/B Dezenfektan Direnç Genlerinin Varligi ve Dezenfektanlara In Vitro Duyarliliklarinin Arastirilmasi

Development of resistance to disinfectant substances in nosocomial microorganisms is an important problem encountered during disinfectant practices. Methicillin-resistant Staphylococcus aureus (MRSA) remains a significant cause of hospital-acquired infections. Besides being resistant to several antimicrobial agents, MRSA strains can also become resistant to some disinfectant substances. Resistance to disinfectant substances may develop due to the misuse of disinfectants. This may either be due to the frequent use of disinfectant substances or use in lower concentrations than recommended. MRSA strains may harbour the qacA/B disinfectant resistance genes that may cause resistance to quarternary ammonium compounds and some cationic disinfectants. These resistance genes are found in plasmids and are responsible for decreased susceptibility or resistance. In this study, a total of 69 nosocomial MRSA strains isolated from clinical specimens in our hospital were tested for disinfectant activity and the presence of qacA/B disinfectant resistance genes in these isolates was investigated by polymerase chain reaction. We determined whether the presence of these genes caused phenotypic resistance to chlorhexidine and benzalkonium chloride by the use of bactericidal and bacteriostatic tests. For this purpose, the minimum inhibitory concentration (MIC) values of these disinfectants against MRSA isolates were detected by microdilution method with the proposals of CLSI, and bactericidal effects of these disinfectants were also detected by using quantitative suspension test according to EN13727:2003 European Standard. It has been found that 11.6% (8/69) of the isolates harbored qacA/B resistance genes. MIC values for chlorhexidine and benzalkonium chloride were found in the range of 2-8 µg/ml. Although it was observed that MIC values were higher in five of the qacA/B gene positive isolates, statistically significant difference was not found between gene positive and gene negative groups. Both 1% chlorhexidine and 1% benzalkonium chloride were found bactericidal against the isolates including the ones carrying the qacA/B resistance genes. It was concluded that the presence of the qacA/B disinfectant resistance genes did not lead to resistance to the disinfectant substances at the concentrations used in clinical practices. Furthermore, tested disinfectants still exhibited bactericidal activity even with high MIC values.

[Investigation of interleukin-10, tumor necrosis factor-alpha and interferon-gamma expression in experimental model of pulmonary aspergillosis]. / Deneysel Pulmoner Aspergilloz Modelinde Interlökin-10, Tümör Nekroz Faktörü-alfa ve Interferon-gama Ifadelenmesinin Arastirilmasi

Pulmonary aspergillosis which is an important opportunistic infection in neutropenic patients, is usually caused by Aspergillus fumigatus. Since the pathogenesis of disease is not well understood, the main proposed mechanism is thought to be cell-mediated immunity and cytokine response. The aim of this study was to investigate the local production of cytokines in the lung tissues of rats with experimentally developed aspergillosis, by reverse transcriptase-polymerase chain reaction (RT-PCR). A total of 33 Wistar albino type rats were included in the study with the consent of Experimental Animal Ethics Committee. Twenty-five of the rats were infected with A.fumigatus by intratracheal way, while 8 animals were used as controls. The presence of A.fumigatus in the lung tissues of infected rats was confirmed with the use of quantitative culture and histologic staining methods. RNA isolation from the lung tissue samples of both groups were performed by a commercial kit (Qiagen, Germany). After obtaining complementary DNAs from the genomic RNAs, in-house qualitative and quantitative (real-time) PCR methods were used to amplify the target regions for interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-?) and interferon-gamma (IFN-?) by using specific primers (Tib Molbiol, Germany). Mean mRNA levels achieved by real-time PCR for IL-10, TNF-? and IFN-? in aspergillosis group were 6.5 x 106 copies/ml, 7.9 x 105 copies/ml and 2.2 x 103 copies/ml, respectively, while those values in control group were 4.3 x 102 copies/ml, 5.6 x 103 copies/ml and 1.3 x 102 copies/ml, respectively. Our data indicated that rat model of aspergillosis was associated with significantly increased expression of mRNA encoding IL-10 and TNF-? than controls (p< 0.05), however there was no statistically significant difference between the groups with respect to IFN-? expression (p= 0.53). In conclusion, the production of proinflammatory cytokines which mediate the influx of phagocytic cells might account for the localization of Aspergillus infection to the upper respiratory tract. The up-regulation of the expression of the immunomodulatory cytokine TNF-? and IL-10 in lung tissue from infected rats might be important to limit the extent of local tissue destruction, but might also account for the fact that infected rats are generally unable to clear the infection spontaneously.

Probiotic agent Saccharomyces boulardii reduces the incidence of lung injury in acute necrotizing pancreatitis induced rats.

BACKGROUND: Acute necrotizing pancreatitis is a severe acute inflammatory disease of the pancreas that can lead to extrapancreatic organ involvement. Supervening lung injury is an important clinical entity determining the prognosis of the patient. Probiotics are dietary supplements known to reduce or alter inflammation and inflammatory cytokines. In the present study, we hypothesize that probiotics may reduce lung injury by reducing bacterial translocation, which results in reduced infection, inflammation, and generation of proinflammatory cytokines in an experimental model of acute necrotizing pancreatitis. METHODS: Pancreatitis was induced by concomitant intravenous infusion of cerulein and glycodeoxycholic acid infusion into the biliopancreatic duct. Saccharomyces boulardii was used as the probiotic agent. Rats were divided into three groups: sham, pancreatitis-saline, which received saline via gavage at 6 and 24 h following the pancreatitis, pancreatitis-probiotics, which received probiotics via gavage method at 6 and 24 h following the pancreatitis. The rats were sacrificed at 48 h, venous blood, mesenteric lymph node, pancreatic and lung tissue samples were obtained for analysis. RESULTS: Serum pancreatic amylase, lactate dehydrogenase, secretory phospholipase A(2), and IL-6 were found to be increased in pancreatitis-saline group compared with the other groups (P < 0.05). Histological analyses revealed that edema, inflammation, and vacuolization as well as polymorphonuclear leukocyte infiltration in the lung tissue was significantly reduced in the probiotic treated group. Bacterial translocation was significantly reduced in the probiotic treated group compared with the other groups (P < 0.05). CONCLUSION: These results suggest that Saccharomyces boulardii reduce the bacterial translocation. As a result of this, reduced proinflammatory cytokines and systemic inflammatory response was observed, which may be the reason underlying reduced lung injury in acute necrotizing pancreatitis.

Determinants of protection against diphtheria in adult hemodialysis patients.

Diphtheria is of great epidemiological concern. Although mainly observed during childhood, unvaccinated adults and relatively immunocompromised patients are at increased risk for acquiring diphtheria. We aimed to determine the rates and certain determinants of protection against diphtheria in adult hemodialysis (HD) patients. Protection rates of 322 HD patients were compared with 65 diabetes mellitus type 2 (DM) patients and 65 healthy controls. A questionnaire was held in regard to smoking habits and alcohol intake. Antibody levels against diphtheria were assessed by an in-house ELISA and a concentration of >or=0.1 IU/mL was regarded as protective. Effects of age, gender and time being on dialysis on protection were assessed by logistic regression. Ratios of individuals with protective antibody levels were found to be 36% (116/322), 27.7% (18/65), and 52.3% (34/65) for HD, DM, and control groups, respectively. Hemodialysis patients had a significantly (p < 0.05) lower protection rate than healthy controls. In all study groups, there was a tendency of higher protection rate with increasing age. These low ratios of protected individuals in both HD and DM patient groups are alarming, as these patients generally have defects in vaccine responses, and carriage is important in the perpetuation of diphtheria. The protection status of these patient groups might be improved with additional vaccinations.