A comparative perspective on brucellar, pyogenic, and tuberculous spondylodiscitis.

OBJECTIVE: Non-specific features of spondylodiscitis lead to a delay and challenge in the diagnosis/differential diagnosis/treatment processes, and thus, serious complications may arise. This study aims to compare brucellar, pyogenic, and tuberculous types of spondylodiscitis, considering their demographic, clinical, and laboratory differences. This may provide more rapid management and good outcomes. PATIENTS AND METHODS: A total of 131 patients with infectious spondylodiscitis were included in the study. The patients were divided into brucellar (n=63), pyogenic (n=53), and tuberculous (n=15) types of spondylodiscitis and compared for demographic, clinical, laboratory, and imaging features. RESULTS: Tuberculous spondylodiscitis had higher scores for weight loss, painless palpation, thoracic spine involvement, and psoas abscess formation than other spondylodiscitis. Also, tuberculous spondylodiscitis had higher rates of neurologic deficit and lower rates of lumbar involvement than brucellar spondylodiscitis. Pyogenic spondylodiscitis is more likely to occur in patients who have a history of spine surgery compared to other forms of spondylodiscitis. Also, pyogenic spondylodiscitis had higher rates of fever, erythema, paraspinal abscess, white blood cell (WBC), and erythrocyte sedimentation rate (ESR) than brucellar spondylodiscitis. On the other hand, brucellar spondylodiscitis had higher rates of rural living and sweating than pyogenic spondylodiscitis. CONCLUSIONS: Weight loss, painless palpation, involved thoracic spine, psoas abscess, and neurologic deficit are symptoms favoring tuberculous spondylodiscitis. History of spine surgery, high fever, skin erythema, and paraspinal abscess are findings in favor of pyogenic spondylodiscitis. Rural living, sweating, and involved lumbar spine are symptoms that indicate brucellar spondylodiscitis. These symptoms can be used to distinguish the types of spondylodiscitis.

An Electron paramagnetic resonance (EPR) spin labeling study in HT-29 Colon adenocarcinoma cells after Hypericin-mediated photodynamic therapy.

BACKGROUND: Colon cancer affects 1.23 million people worldwide and is the third most common malignant disease in men and the second in women. The only curative treatment is surgical resection, but a significant number of patients develop local recurrence or distant metastases. One of the alternative treatment methods for colon cancer is photodynamic therapy (PDT). In recent years, hypericin (HYP) derived from Hypericum perforatum has been suggested as a strong candidate photosensitizer for PDT. Our interest is focused on the biophysical changes in colon cancer cells in relation to HYP-mediated PDT. RESULTS: In this study, HYP-mediated PDT at 0.04, 0.08 or 0.15 µM HYP concentrations was performed in HT-29 colon adenocarcinoma cells and the Electron Paramagnetic Resonance (EPR) spectra of the spin labeled cells were obtained. Plasma membranes are already heterogeneous structures; the presence of cancer cells increased the heterogeneity and also fluidity of the plasma membranes. Therefore, the obtained spectra were evaluated by EPRSIMC program, which provides the calculation of heterogeneous structures up to four spectral components with different fluidity characteristics. Generally, two motional patterns were obtained from calculations and the number of them increased at the highest concentration. As the order parameters of the most populated components compared, an increase was observed depending on the HYP concentration. However, because of the heterogeneous structure of membrane, the order parameters of the less populated components did not exhibit a regular distribution. CONCLUSION: After HYP-mediated PDT, concentration dependent changes were observed in the domain parameters indicating an increase in the HYP accumulation.

Serum paraoxonase activity and oxidative stress levels in patients with cutaneous anthrax.

INTRODUCTION: Anthrax is a bacterial disease caused by the aerobic sporeforming bacterium Bacillus anthracis. It has been suggested that oxidative stress plays an important role in the pathogenesis of B. anthracis. The aim of this study was to investigate serum paraoxonase 1 (PON1) activity, catalase activity, malondialdehyde (MDA) levels, and superoxide dismutase (SOD) levels in patients with cutaneous anthrax. MATERIALS AND METHODS: Fifteen patients with cutaneous anthrax and 15 healthy controls were enrolled in this study. The serum MDA levels, SOD levels, paraoxonase, arylesterase, and catalase activities were measured using a spectrophotometer. RESULTS: The serum SOD levels, paraoxonase, arylesterase, and catalase activities were significantly lower in patients with cutaneous anthrax than in controls (for all, p < 0.001), whereas MDA levels were significantly higher ( p < 0.001). No significant correlation was found between serum paraoxonase activity, arylesterase activity, SOD levels, and MDA levels (all, p > 0.05) in patients with cutaneous anthrax. CONCLUSIONS: The current study was the first to show decreased antioxidant levels and increased oxidant levels in patients with cutaneous anthrax. Therefore, decreased PON1 activity may play a role in the pathogenesis of cutaneous anthrax.

Effect of cis-(Z)-flupentixol on DPPC membranes in the presence and absence of cholesterol.

Cis-(Z)-flupentixol dihydrochloride (FLU), a thioxanthene drug, is used in therapy of schizophrenia as well as in anxiolytic and depressive disorders. Since the action mechanism of FLU is not completely understood, the main objective of present study is to provide a detailed evaluation of flupentixol-phospholipid membrane interactions at molecular level. FLU-dipalmitoylphosphatidylcholine (DPPC) interactions in presence and absence of cholesterol (CHO) were investigated as a function of temperature. The changes in upper part of membrane were more pronounced than those in central part of membrane, as indicated by EPR and FTIR. FLU was proposed to incorporate into phospholipid membranes with its triple ring parallel to head group and its chain toward alkyl chain of phospholipids. According to DSC results, the incorporation of 10 mol% FLU into DPPC caused a shoulder in transition peak, suggesting the occurence of a phase separation, and formation of this new phase is still observable in presence of CHO. It is well known that, structure and dynamics of lipids have significant influence on the function of membrane bound proteins, and consecutively their actions. Based upon these, it was proposed that FLU may modify membrane associated receptors and transport proteins, which would form the basis of its clinical efficiency.

Spectroscopic and calorimetric studies on trazodone hydrochloride-phosphatidylcholine liposome interactions in the presence and absence of cholesterol.

The interaction of antidepressant drug trazodone hydrochloride (TRZ) with dipalmitoyl phosphatidylcholine (DPPC) multilamellar liposomes (MLVs) in the presence and absence of cholesterol (CHO) was investigated as a function of temperature by using Electron Paramagnetic Resonance (EPR) spin labeling, Fourier Transform Infrared (FTIR) Spectroscopy and Differential Scanning Calorimetry (DSC) techniques. These interactions were also examined for dimyristoyl phosphatidylcholine (DMPC) multilamellar liposomes by using Electron Paramagnetic Resonance (EPR) spin labeling technique. In the EPR spin labeling studies, 5- and 16-doxyl stearic acid (5-DS and 16-DS) spin labels were used to monitor the head group and alkyl chain region of phospholipids respectively. The results indicated that TRZ incorporation causes changes in the physical properties of PC liposomes by decreasing the main phase transition temperature, abolishing the pre-transition, broadening the phase transition profile, and disordering the system around the head group region. The interaction of TRZ with unilamellar (LUV) DPPC liposomes was also examined. The most pronounced effect of TRZ on DPPC LUVs was observed as the further decrease of main phase transition temperature in comparison with DPPC MLVs. The mentioned changes in lipid structure and dynamics caused by TRZ may modulate the biophysical activity of membrane associated receptors and in turn the pharmacological action of TRZ.

Genitourinary brucellosis: results of a multicentric study.

This study reviewed the clinical, laboratory, therapeutic and prognostic data on genitourinary involvement of brucellosis in this largest case series reported. This multicentre study pooled adult patients with genitourinary brucellar involvement from 34 centres treated between 2000 and 2013. Diagnosis of the disease was established by conventional methods. Overall 390 patients with genitourinary brucellosis (352 male, 90.2%) were pooled. In male patients, the most frequent involved site was the scrotal area (n=327, 83.8%), as epididymo-orchitis (n=204, 58%), orchitis (n=112, 31.8%) and epididymitis (n=11, 3.1%). In female patients, pyelonephritis (n=33/38, 86.8%) was significantly higher than in male patients (n=11/352, 3.1%; p<0.0001). The mean blood leukocyte count was 7530±3115/mm3. Routine laboratory analysis revealed mild to moderate increases for erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). The mean treatment duration and length of hospital stay were significantly higher when there were additional brucellar foci (p<0.05). Surgical operations including orchiectomy and abscess drainage were performed in nine (2.3%) patients. Therapeutic failure was detected in six (1.5%), relapse occurred in four (1%), and persistent infertility related to brucellosis occurred in one patient. A localized scrotal infection in men or pyelonephritis in women in the absence of leucocytosis and with mild to moderate increases in inflammatory markers should signal the possibility of brucellar genitourinary disease.

Liver involvement in patients with brucellosis: results of the Marmara study.

Brucellosis is a zoonotic disease that primarily affects the reticuloendothelial system. But, the extent of liver damage in due course of the disease is unclear. This study included 325 brucellosis patients with significant hepatobiliary involvement identified with microbiological analyses from 30 centers between 2000 and 2013. The patients with ≥5 times of the upper limit of normal for aminotransferases, total bilirubin level ≥2 mg/dl or local liver lesions were enrolled. Clinical hepatitis was detected in 284 patients (87.3 %) and cholestasis was detected in 215 (66.1 %) patients. Fatigue (91 %), fever (86 %), sweating (83 %), arthralgia (79 %), and lack of appetite (79 %) were the major symptoms. Laboratory tests showed anemia in 169 (52 %), thrombocytopenia in 117 (36 %), leukopenia in 81 (25 %), pancytopenia in 42 (13 %), and leukocytosis in 20 (6 %) patients. The most commonly used antibiotic combinations were doxycycline plus an aminoglycoside (n = 73), doxycycline plus rifampicin (n = 71), doxycycline plus rifampicin and an aminoglycoside (n = 27). The duration of ALT normalization differed significantly in three treatment groups (p < 0.001). The use of doxycycline and an aminoglycoside in clinical hepatitis showed better results compared to doxycycline and rifampicin or rifampicin, aminoglycoside, doxycycline regimens (p < 0.05). However, the length of hospital stay did not differ significantly between these three combinations (p > 0.05). During the follow-up, treatment failure occurred in four patients (1 %) and relapse was seen in three patients (0.9 %). Mortality was not observed. Hepatobiliary involvement in brucellosis has a benign course with suitable antibiotics and the use of doxycycline and an aminoglycoside regimen seems a better strategy in select patients.

Interaction of antidepressant drug, clomipramine, with model and biological stratum corneum membrane as studied by electron paramagnetic resonance.

The interactions of tricyclic antidepressant drug, clomipramine (CLO), with pig ear stratum corneum (SC) and model membranes were investigated by electron paramagnetic resonance (EPR) spin labeling to get some insight into the possible application of this drug in transdermal delivery. The changes in membrane characteristics caused by CLO in the regions that are close to the water-lipid interfaces and the central parts of the membranes were searched. The experimental results were supported by computer simulation of EPR spectra, which showed heterogeneity of the membranes composed of regions with different fluidity characteristics. CLO was effective in both parts of the layers, indicating intercalation of the drug into model membranes as well as into the pig ear SC. Introduction of various molar ratios of CLO caused a decrease in the order parameter and an increase in the rotational diffusion of nitroxide moiety in different membrane regions as well as an increase in the polarity of spin probe environment. It also changed the number of resolved spectral components, which reflects the heterogeneity of the membrane. The fluidizing effect of CLO on pig ear SC throughout the whole membrane layers indicates that CLO penetrates into the SC, which is important for its transdermal delivery.

Association of paraoxonase activity and atherosclerosis in patients with chronic hepatitis B.

BACKGROUND: The hepatitis B virus is a significant pathogen that causes cirrhosis, and hepatocellular cancer as a result of the damage it causes to liver cells. Its infection affects more than 400 million people globally. Although there is an effective vaccine and treatment methods, almost 1, 000, 000 people die every year. OBJECTIVE: To investigate paraoxonase and arylesterase activities along with oxidative status parameters and serum lipid levels, and to find out if there is any increased susceptibility to atherogenesis. METHODS: Thirty-four subjects with chronic hepatitis B and 39 healthy subjects as control were enrolled in the study. Age, body mass index and gender, Serum Triglycerides (TG), High-density Lipoprotein (HDL) and Low-Density lipoprotein (LDL) levels, serum paraoxonase-1 and arylesterase activities were determined. Oxidative and antioxidative statuses were evaluated by measuring serum-free sulfhydryl groups, lipid hydroperoxide levels, total antioxidant capacity, total oxidant status, and oxidative stress index. RESULTS: Serum TG and LDL levels were higher while serum HDL levels were lower in patients with chronic hepatitis B than in controls but the differences did not reach statistical significance. Serum paraoxonase-1 and arylesterase activities, plasma free sulfhydryl groups, and total antioxidant capacity were significantly lower in patients than in controls (p=0.018, p=0.005, p<0.001, p=0.037 respectively), while lipid hydroperoxide, total oxidant status, and oxidative stress index were significantly higher (for all p<0.001). CONCLUSION: The diminution in the paraoxonase-1 and arylesterase activities could contribute to the accelerated development of atherosclerosis in patients with chronic hepatitis B.

Evaluation of oxidative status in patients with Fasciola hepatica infection.

BACKGROUND: F. hepatica infection is rare and mostly subclinical. Migration of juvenile forms of F. hepatica into the host's liver is accompanied by an inflammatory reaction followed by fibrosis and cirrhosis. OBJECTIVE: The aim of this study was to evaluate oxidative status by using a novel automated method in patients with Fasciola hepatica. METHODS: Twenty two patients with a diagnosis of F. hepatica and 26 healthy volunteers were enrolled in the study. Their Total antioxidant capacity status (TAC), total oxidant status (TOS) and catalase were measured in them and oxidative stress index (OSI) was calculated. These measurements were also taken for the control group and the values were compared. RESULTS: Plasma levels of total TOS and OSI were significantly increased in patients as compared with healthy controls (P<0.001, P<0.001, P=0.001, P=0.008) respectively. In contrast, TAC level was significantly lower in patients as compared with controls (p<0.05). There was no significant difference between the catalase results of the two groups (p>0.05). CONCLUSION: Total oxidative status and OSI were increased and total antioxidative status capacity was decreased in patients with F. hepatica infection. A high oxidative stress occurs during F. hepatica infection, which may cause severe damage in both the liver.

EPR investigation of clomipramine interaction with phosphatidylcholine membranes in presence and absence of cholesterol.

The effects of tricylic antidepressant clomipramine (CLO) on the membrane properties of saturated dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine as well as on unsaturated egg yolk phosphatidylcholine liposomes were investigated by the electron paramagnetic resonance spin-labeling technique, in combination with the simulation of the spectra, taking into account that the membrane is heterogeneous and composed of the regions with different fluidity characteristics. Different spin labels, monitoring membrane properties in the upper and inner parts of the membrane, were used. In general, two spectral components, having different motional characteristics, were detected in all liposomes investigated. In liposomes with saturated chains, CLO decreased the phase-transition temperature, disordered the membrane, and increased polarity in the upper part of the membrane. However, less impact was observed in liposomes with unsaturated chains. In dipalmitoyl phosphatidylcholine liposomes, it also induced molecular rearrangements near the pretransition temperature. The presence of 30 mol% cholesterol increased the fluidizing effect of CLO and modified the lateral diffusion of nitroxide in the inner part of the membrane. A unique anomalous increase in diffusion of nitroxide, dependent on CLO concentration, was detected in the temperature region where the phosphatidylcholine membrane without cholesterol experiences the phase transitions. Since the changes in the central part of the membrane were even more pronounced than in the upper part of the membrane, it could be concluded that CLO incorporates into the membrane with its hydrophobic ring parallel to the phospholipid chains.

EPR spin label study of walnut oil effects on phosphatidylcholine membranes.

Effects of walnut oil (WO) on dynamic and thermodynamic properties of 0-50 wt% cholesterol (CH) containing dimyristoylphosphatidylcholine (DMPC) and 10 wt% CH containing dipalmitoylphosphatidylcholine (DPPC) membrane dispersions were studied by electron paramagnetic resonance (EPR), using 5-doxyl stearic acid (5-DSA) and 16-doxyl stearic acid (16-DSA). Incorporation of 10 wt% WO alone decreased the phase transition temperature and created depth-dependent effects at the gel phase. The order increased close to the head region and decreased in the hydrocarbon core of the DMPC bilayer. For DPPC, the order decreased both close to head region and in the hydrocarbon core. Ten weight percent WO did not have considerable effect at the fluid phase for both DMPC and DPPC. Incorporation of 40 wt% WO into DMPC created an abrupt decrease in the maximum hyperfine splitting values after 305 K. The effect of 10 wt% WO in CH containing DMPC dispersions was dependent on the CH concentration. An increase and a decrease in the order were observed at low and high CH concentrations, respectively. Incorporation of WO created different effects on fluidity of 10 wt% CH containing DMPC and DPPC dispersions. Close to the head group region, the order in DMPC increased both in the gel and fluid phases; but for DPPC, an opposite effect was observed in both of the phases. In the hydrocarbon core of the bilayer, addition of 10 wt% WO into 10 wt% CH containing DMPC decreased the order in the gel phase and WO did not affect the order in the fluid phase. For DPPC, WO effects were observed to alter with temperature. In the studied temperature range, order parameters, diffusion constants and effective tilt angles were obtained from simulations of the spectra using Microscopic Order Macroscopic Disorder (MOMD) and Vary Anisotropic Reorientation (VAR) models. For 16-DSA, spectra were also simulated using two domains with EPRSIM.

The use of simulation in the EPR spin probe technique for detection of irradiated seeds.

An electron paramagnetic resonance (EPR) spin probe study of irradiated wheat seeds was performed depending on irradiation dose. The structural changes in the membrane integrity were followed using aqueous solutions of 4-hydroxy-TEMPO (TANOL) spin probe and a line broadening material. In the studies dry seed embryos were kept in these solutions for 150 min. The spectra were recorded at various times of air drying process. The simulation of these spectra indicated a decrease in the water content of the embryos depending on the increasing irradiation dose. This indicates the increase in the permeability of the membranes as a result of the radiation damage. From the decay curves it is possible to determine about irradiation dose, however, this approach is not very successful at close irradiation doses.

A DNP investigation of pi and final sigma transitions for nitroxide spin probes at 1.53 mT.

A weak field DNP study of 4-hydroxy-2,2,6, 6-tetramethyl-1-piperidinyloxy (4-hydroxy-TEMPO), 4-amino-2,2,6, 6-tetramethyl-1-piperidinyloxy (4-amino-TEMPO), and 2, 2-Di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) free radicals invarious solvents was performed at 1.53 mT using a doubleresonance spectrometer. pi transitions, in which the RF fieldhas a component perpendicular to the static magnetic fieldB0, and final sigma transitions, in which the RF field has a component parallel to B0, were recorded. In order to observe thefinal sigma transitions a new probe head was designed. The final sigma transitions were obtained at the frequencies expected from theoretical calculations. However, the results obtained for the observed enhancements deviated from the values expectedtheoretically.