Diagnostic potential of Brucella melitensis Rev1 native Omp28 precursor in human brucellosis.

Serologic tests for brucellosis aim to detect antibodies produced against membranous lipopolysaccharide of bacteria. Diagnostic use of this method is limited due to false positiveness. This study evaluates an alternative antigen to lipopolysaccharides (LPS), outer membrane 28-precursor-protein, of Brucella melitensis Rev1 for its diagnostic value. Omp28 precursor of B. melitensis Rev1 was cloned, expressed, and purified. 6-His and sumo epitope tags were used to tag the protein at N-termini. Omp28 gene was amplified based on the ORF sequence and cloned into a pETSUMO vector. The recombinant construct was propagated in Escherichia coli One Shot® Mach1™ cells then transformed into E. coli BL21(D3) cells for protein expression. The purified protein was studied in an indirect ELISA for diagnosis of brucellosis. Sera samples from 60 patients were screened by ELISA and the results were compared to Rose Bengal plate test. Recombinant antigen-based iELISA has given a successful outcome with the sensitivity, specificity, positive predictive value, and negative predictive value of 87.8%, 96.2%, 96.6%, and 78.78%, respectively. In conclusion, recombinant production and purification of the immunodominant Omp28 precursor protein has been achieved successfully in a one-step process with efficient yield and can be used for diagnosis of brucellosis in humans.

Cranial imaging findings in neurobrucellosis: results of Istanbul-3 study.

OBJECTIVE: Neuroimaging abnormalities in central nervous system (CNS) brucellosis are not well documented. The purpose of this study was to evaluate the prevalence of imaging abnormalities in neurobrucellosis and to identify factors associated with leptomeningeal and basal enhancement, which frequently results in unfavorable outcomes. METHODS: Istanbul-3 study evaluated 263 adult patients with CNS brucellosis from 26 referral centers and reviewed their 242 magnetic resonance imaging (MRI) and 226 computerized tomography (CT) scans of the brain. RESULTS: A normal CT or MRI scan was seen in 143 of 263 patients (54.3 %). Abnormal imaging findings were grouped into the following four categories: (a) inflammatory findings: leptomeningeal involvements (44), basal meningeal enhancements (30), cranial nerve involvements (14), spinal nerve roots enhancement (8), brain abscesses (7), granulomas (6), and arachnoiditis (4). (b) White-matter involvement: white-matter involvement (32) with or without demyelinating lesions (7). (c) Vascular involvement: vascular involvement (42) mostly with chronic cerebral ischemic changes (37). (d) Hydrocephalus/cerebral edema: hydrocephalus (20) and brain edema (40). On multivariate logistic regression analysis duration of symptoms since the onset (OR 1.007; 95 % CI 1-28, p = 0.01), polyneuropathy and radiculopathy (OR 5.4; 95 % CI 1.002-1.013, p = 0.044), cerebrospinal fluid (CSF)/serum glucose rate (OR 0.001; 95 % CI 000-0.067, p = 0.001), and CSF protein (OR 2.5; 95 % CI 2.3-2.7, p = 0.0001) were associated with diffuse inflammation. CONCLUSIONS: In this study, 45 % of neurobrucellosis patients had abnormal neuroimaging findings. The duration of symptoms, polyneuropathy and radiculopathy, high CSF protein level, and low CSF/serum glucose rate were associated with inflammatory findings on imaging analyses.

Results of a multinational study suggest the need for rapid diagnosis and early antiviral treatment at the onset of herpetic meningoencephalitis.

Data in the literature regarding the factors that predict unfavorable outcomes in adult herpetic meningoencephalitis (HME) cases are scarce. We conducted a multicenter study in order to provide insights into the predictors of HME outcomes, with special emphasis on the use and timing of antiviral treatment. Samples from 501 patients with molecular confirmation from cerebrospinal fluid were included from 35 referral centers in 10 countries. Four hundred thirty-eight patients were found to be eligible for the analysis. Overall, 232 (52.9%) patients experienced unfavorable outcomes, 44 died, and 188 survived, with sequelae. Age (odds ratio [OR], 1.04; 95% confidence interval [CI], 1.02 to 1.05), Glasgow Coma Scale score (OR, 0.84; 95% CI, 0.77 to 0.93), and symptomatic periods of 2 to 7 days (OR, 1.80; 95% CI, 1.16 to 2.79) and >7 days (OR, 3.75; 95% CI, 1.72 to 8.15) until the commencement of treatment predicted unfavorable outcomes. The outcome in HME patients is related to a combination of therapeutic and host factors. This study suggests that rapid diagnosis and early administration of antiviral treatment in HME patients are keys to a favorable outcome.

Determination of Serum Differential Carnitine Ester Levels in HIV(+) Patients: A Cross-Sectional Study.

OBJECTIVE: It has been reported that carnitine deficiency is observed in various viral infections and in the follow-up of the prognosis of some diseases. In this cross-sectional study, we aimed to determine how carnitine ester derivatives change in HIV-positive patients. MATERIALS AND METHODS: In this study, 25 HIV-infected patients who applied to Harran University Faculty of Medicine Education Research and Practice Hospital Infectious Diseases and Clinical Microbiology Outpatient Clinic and who did not receive any antiretroviral treatment, as well as 25 healthy volunteers were included in the study. Carnitine ester levels in serum samples were measured by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) method (Shimadzu North America, Columbia, MD, USA). RESULTS: While suberoylcarnitine (C8DC), myristoleylcarnitine (C14:1), tetradecadienoylcarnitine (C14:2), palmitoleylcarnitine (C16:1), and linoleylcarnitine (C18:2) levels in HIV(+) patients were quite low compared to the control group, tiglylcarnitine (C5:1) levels were high (p ≤ 0.05). In addition, C5:1 and C14:2 index parameters according to VIP score, and C5:1 and C14:1/C16 index parameters according to ROC analysis were determined as markers with high potential to distinguish HIV(+) patients from healthy volunteers. CONCLUSION: This study showed that levels of acylcarnitine derivatives might be altered in HIV(+) patients, and the results obtained may contribute to a better understanding of carnitine metabolism.

Evaluation of cerebrospinal fluid levels for ALOX5, S100B, DEFA1, and GFAP in infectious meningitis.

BACKGROUND: The aim of this study was to determine how the levels of peptide and protein-based biomarkers in cerebrospinal fluid change in bacterial, tuberculous, and aseptic meningitis, and to determine the success of these agents in distinguishing between different types of infectious meningitis. METHODS: The levels of arachidonate-5-lipoxygenase, S100 calcium-binding protein B, defensin-α 1, and glial fibrillary acidic protein in cerebrospinal fluid samples from 20 tuberculosis, 40 bacterial, 25 aseptic meningitis patients, and 55 control groups were measured and compared using an enzyme-linked immunosorbent assay. RESULTS: The mean age of the patients was 37.9 ±â€…14.4 years. The parameter that contributed the most to the differential diagnosis of the infectious meningitis groups was S100 calcium-binding protein B. The S100 calcium-binding protein B levels were significantly higher in the tuberculous meningitis group than in the other groups, and arachidonate-5-lipoxygenase levels were significantly higher in the tuberculous meningitis and bacterial meningitis groups (P < .05). CONCLUSION: This study showed that cerebrospinal fluid arachidonate-5-lipoxygenase, and S100 calcium-binding protein B levels may differ in bacterial, aseptic, and tuberculous meningitis, and the results obtained may be quite effective as important potential biomarkers in the differential diagnosis of different types of meningitis.

The activity of paraoxonase and arylesterase in patients with osteomyelitis.

BACKGROUND: The aim of this study was to evaluate oxidative stress and to determine the activity of paraoxonase and arylesterase in patients with osteomyelitis compared to healthy controls. METHOD: In total, 30 patients diagnosed with osteomyelitis and 30 healthy volunteers were enrolled in the study. Paraoxonase and arylesterase activities were measured spectrophotometrically. Serum lipid hydroperoxide (LOOH) concentrations were measured by ferrous oxidation with xylenol orange (FOX) assay as markers of oxidative stress. RESULTS: Serum paraoxonase and arylesterase activities were significantly lower in patients with osteomyelitis compared to control individuals (all p < 0.05). Serum LOOH concentrations were significantly higher in patients with osteomyelitis than those in controls (p < 0.05). Arylesterase activity was inversely correlated with triglyceride (r =- 0.49; p = 0.005) and cholesterol concentrations (r =- 0.41; p = 0.025). CONCLUSION: In light of the findings obtained from the present study, it may be assumed that decreased activity of serum paraoxonase and increased concentrations of LOOH observed in osteomyelitis patients appear to be related to the increased oxidative stress and inflammatory conditions present in these patients, and may cause a much more severe status of the disease.

Prolidase and oxidative stress in chronic hepatitis C.

BACKGROUND: Hepatitis C infection represents a common healthcare issue worldwide. The present trial was designed to investigate the role of prolidase, an enzyme that is significantly involved in the biosynthesis of collagen, and of the oxidative stress that is considered to be involved in the pathogenesis of various diseases, in the chronic hepatitis C infection. The trial was performed to assess the serum prolidase enzyme level and the oxidative-antioxidative status and to determine the relation between the serum prolidase activity and the oxidative stress parameters. METHODS: A total of 95 individuals, including 55 patients with chronic hepatitis C infection (CHC) and 40 healthy adults, were enrolled in the trial. The values for prolidase, the total antioxidant status (TAS), the total oxidative stress (TOS), the oxidative stress index (OSI), sulfhydryl (SH), lipid peroxidation LOOH, catalase (CAT), and ceruloplasmin were measured and compared between the patient groups. RESULTS: The prolidase, TOS, LOOH, CAT, and the OSI values were higher in the chronic hepatitis C group compared to the control group (P < 0.001). The TAS, SH, and ceruloplasmin levels were lower in the CHC group relative to the control group (P < 0.001). CONCLUSION: We suppose that the values of prolidase and the oxidative stress are increased while the antioxidant levels are decreased in CHC. As a result, prolidase and the oxidative stress seem to be related with the progression of the disease.

[Can mannose-binding lectin and plasma level of soluble urokinase receptor be used in diagnosis and treatment monitorization of brucellosis patients?]. / Brusellozlu Hastalarda Mannoz-Baglayan Lektin ve Plazma Çözünür Ürokinaz Reseptör Düzeyi Tani ve Tedavi Izleminde Kullanilabilir mi?

The aim of this study was to evaluate the diagnostic value of serum mannose-binding lectin (MBL) and plasma soluble urokinase plasminogen activator receptor (SuPAR) levels in monitoring the treatment in patients with brucellosis, by comparing their levels before and after treatment with the values obtained from healthy control group. Thirty brucellosis patients (mean age: 25.8 ± 12.2 years; 15 were male) and 28 healthy controls (mean age: 29.3 ± 12.3 years; 15 were male) were included in the study. Patients were diagnosed with brucellosis according to the characteristic clinical findings and by brucella standard tube agglutination test (SAT) titer ≥ 1/160 and/or blood culture positivity. Serum MBL (Antibodyshop, Denmark) and plasma SuPAR (Virogates, Denmark) levels were investigated with commercial ELISA kits. In our study, no statistical significance was observed between the pre-treatment (13.8 ± 13.4 ng/ml) and post-treatment (12.4 ± 13.1 ng/ml) MBL levels of the patient group and MBL levels of the control group (16.5 ± 14.8 ng/ml) (p> 0.05). Moreover, the mean SuPAR levels measured in pre-treatment and post-treatment plasma samples of the brucellosis patients was 5.1 ± 1.9 ng/ml and 2.9 ± 1.3 ng/ml, respectively, while the mean SuPAR level was 1.8 ± 0.5 ng/ml in the control group. The difference between mean SuPAR levels of patients in pre- and post-treatment samples was found statistically significant (p< 0.001). In addition SuPAR levels were significantly higher in patients before and after treatment than the control group (p> 0.001). In conclusion, plasma SuPAR level would be a useful marker for the diagnosis and treatment follow up of the patients with brucellosis.

Plasma Free Amino Acid Profile in HIV-Positive Cases.

BACKGROUND: Increasing the sensitivity and availability of liquid chromatography tandem mass spectrometry (LC-MS/MS) devices may provide advantages in terms of revealing the changes in metabolic pathways in HIV-positive patients and elucidating the physiopathology. INTRODUCTION: The aim of this study was to determine the difference in amino acid levels between HIV-positive patients and healthy individuals by using LC-MS / MS and investigate its relationship with HIV infection. MATERIAL AND METHODS: Concentrations of 36 different amino acids and their derivatives were measured and compared in venous plasma samples from 24 HIV-positive patients and 24 healthy individuals by using the LC-MS/MS method (Shimadzu North America, Columbia, MD, USA). RESULTS: HIV-positive subjects had significantly lower alanine, 1-methyl-L-histidine, valine, aspartate, cysteine, cystine, methionine, lysine, glutamine, imino acid, tyrosine, tryptophan, threonine, sarcosine, and argininosuccinic acid and significantly higher 3-methyl-L -histidine, asparagine, glutamate, and carnosine levels as compared to healthy controls. No significant differences were detected in other amino acids. CONCLUSION: The significant differences in amino acid profile between HIV-positive and healthy subjects may represent an auxiliary biomarker of cellular damage in asymptomatic HIV-positive patients that may be examined in more detail in further studies. It may also provide guidance for symptomatic cases in terms of the association between symptoms, clinical manifestations, and deficiency or excess of certain amino acids in the context of the complete metabolomics record of HIVpositive patients.

Efficacy and Safety of Direct-Acting Antivirals in Elderly Patients with Chronic Hepatitis C: A Nationwide Real-Life, Observational, Multicenter Study from Turkey.

BACKGROUND: The number and proportion of elderly patients living with chronic hepatitis C are expected to increase in the coming years. We aimed to compare the real-world efficacy and safety of direct-acting antiviral treatment in elderly and younger Turkish adults infected with chronic hepatitis C. METHODS: In this multicenter prospective study, 2629 eligible chronic hepatitis C patients treated with direct-acting antivirals between April 2017 and December 2019 from 37 Turkish referral centers were divided into 2 age groups: elderly (≥65 years) and younger adults (<65 years) and their safety was compared between 2 groups in evaluable population. Then, by matching the 2 age groups for demographics and pretreatment risk factors for a non-sustained virological response, a total of 1516 patients (758 in each group) and 1244 patients (622 in each group) from the modified evaluable population and per-protocol population were included in the efficacy analysis and the efficacy was compared between age groups. RESULTS: The sustained virological response in the chronic hepatitis C patients was not affected by the age and the presence of cirrhosis both in the modified evaluable population and per-protocol population (P = .879, P = .508 for modified evaluable population and P = .058, P = .788 for per-protocol population, respectively). The results of the per-protocol analysis revealed that male gender, patients who had a prior history of hepatocellular carcinoma, patients infected with non-genotype 1 hepatitis C virus, and patients treated with sofosbuvir+ribavirin had a significantly lower sustained virological response 12 rates (P < .001, P = .047, P = .013, and P = .025, respectively). CONCLUSION: Direct-acting antivirals can be safely used to treat Turkish elderly chronic hepatitis C patients with similar favorable efficacy and safety as that in younger adults.