Determination of phenolic compounds in human saliva after oral administration of red wine by high performance liquid chromatography.

Red wine is a relevant source of bioactive compounds, which contribute to its antioxidant activity and other beneficial advantages for human health. However, the bioavailability of phenols in humans is not well understood, and the inter-individual variability in the production of phenolic compounds has not been comprehensively assessed to date. The present work describes a new method for the extraction and analysis of phenolic compounds including gallic acid (Gal), vanillic acid (Van), caffeic acid (Caf), syringic acid (Sir); (-)-epicatechin (Epi); p-coumaric acid (Cum) and resveratrol (Rsv) in human saliva samples. The target analytes were extracted using Fabric Phase Sorptive Extraction (FPSE), and subsequently analysed by high-performance liquid chromatography (HPLC) coupled with photodiode array detector (PDA). Chromatographic separation was achieved using a Symmetry C18 RP column in gradient elution mode, with methanol and phosphate buffer as the mobile phases. The linearity (intercept, slope, and determination coefficient) was evaluated in the range from 1 to 50 µg/mL. The limit of quantification (LOQ) was 1 µg/mL (LLOQ ≥0.8 µg/mL), whereas limit of detection was 0.25 µg/mL. The intra and inter-day RSD% and BIAS% values were less than± 15%. The analytical performances were further tested on human saliva collected from healthy volunteers after administering red wine. To the best of our knowledge, this is the first FPSE procedure for the analysis of phenols in saliva, using a non-invasive and easy to perform sample collection protocol. The proposed fast and inexpensive approach can be deployed as a reliable tool to study other biological matrices to proliferate understanding of these compounds distribution in human body.

Fast off-line FPSE-HPLC-PDA determination of six NSAIDs in saliva samples.

A fast off-line FPSE-HPLC-PDA method has been reported that allows simultaneous clean up and determination of six non-steroidal anti-inflammatory drugs (NSAIDs) in saliva samples from healthy volunteers. Particularly, furprofen, indoprofen, ketoprofen, fenbufen, flurbiprofen, and ibuprofen were chromatographically resolved. Benzyl paraben was chosen as the internal standard (BzPB, IS). These target compounds were successfully extracted from human saliva using fabric phase sorptive extraction (FPSE) and then analysed in the liquid chromatographic system by means of a short analytical column (Symmetry C18, 75 × 4.6 mm, 3.5 µm) using acetonitrile (AcN) and phosphate buffer (PBS, 30 mM; pH = 2.5) as the mobile phases. The method, validated through the calculation of all analytical parameters in accordance of International Guidelines, was applied to real saliva sample analysis collected from informed volunteers. The proposed approach that included the use of sol-gel polytetrahydrofuran (sol-gel PTHF) sorbent immobilized on cellulose support and C18 stationary phase used in HPLC, showed high potential as a fast tool for future clinical and forensic applications. The herein reported results encourage potential future application of FPSE in the forensic field. Furthermore, the FPSE membrane was tested in dried saliva spot mode (DSS) in order to check its potential use as a sampling device, also for forensic applications.

Biofluid sampler: A new gateway for mail-in-analysis of whole blood samples.

The current study reports the development of a novel biofluid sampler (BFS) which is capable of sampling and sample preparation of whole blood without converting it into plasma or serum. The sampler can retain a whole blood sample from 10 to 1000 µL. Although the device shares the same working principle of dried blood spot (DBS) cards, it eliminates most of the technological shortcomings of DBS cards such as low maximum sample volume (~50 µL), sample inhomogeneity due to haematocrit, and poor physical adsorption driven analyte retention by incorporating sol-gel derived high efficiency, multi-functional sorbents on cellulose fabric substrate. The performance of BFS was tested via "Mail-in-Analysis" using three non-steroidal anti-inflammatory drugs (NSAIDs, ketoprofen, carprofen and diclofenac) as the test compounds. Human whole blood samples were fortified with the test compounds and sampled on conventional DBS cards and biofluid samplers (BFSs) in the USA. After drying the blood samples at room temperature, the samples were shipped to Italy for chromatographic analysis. The analytes were back-extracted from the DBS cards and BFSs using methanol and subsequently analysed using a short Symmetry C18 column (75 × 4.6 mm, 3.5 µm). Acetonitrile (ACN) and PBS (30 mM; pH = 2.5) were used as the mobile phases and the elution was performed under isocratic conditions. Compared to the classical dried blood spot cards (DBS), BFSs offer better performance in retaining the selected NSAIDs under conventional postal shipment. By substantially expanding the sampling capacity, eliminating most of the shortcomings of classical DBS cards and exploiting the better materials properties of sol-gel based functional sorbents, BFSs offer a new and profoundly simplified approach for whole blood sampling and analysis and is expected to change the current practice of blood analysis, allowing accurate quantitative analyses either in a local laboratory (on site) or using mail-in-analysis (off site) without compromising the quality of bioanalytical data.

Intraperitoneal Administration of Low Dose Aluminium in The Rat: How Good is It to Produce a Model for Alzheimer Disease.

Since neurotoxicity of aluminium (Al) resembles the progressive neurodegeneration observed in Alzheimer Disease (AD), Al administration in several ways has been used to produce AD model. Intraperitoneal (ip) low dose (4.2 mg/ kg) Al injection in rats for long periods is the preferred method by some researchers. In this paper, the efficiency of this method for producing an AD model was evaluated. In this study, we looked at the neuropathology of Al and the characteristic lesions of AD by histological and immunohistochemical techniques and determined oxidative stress markers in the brains of Al-treated and control rats. We also made electrophysiological recordings at the hippocampus and evaluated possible behavioural changes by Morris water maze test. However, no pathologic changes occurred in the animals except for an impairment in long-term potentiation (LTP) in the hippocampus (e.g. the LTPs of population spike (PS) amplitude at 15 min post-tetanus were measured as 217±27% in Al-treated rats and as 240±42% in sham-treated rats, of baseline PS amplitude). According to the findings of the present study, low dose of ip Al in rats is not sufficient to produce a good AD model. Higher doses (≥10 mg/kg) should be used.

Effectiveness of a multidimensional approach for prevention of ventilator-associated pneumonia in 11 adult intensive care units from 10 cities of Turkey: findings of the International Nosocomial Infection Control Consortium (INICC).

PURPOSE: To evaluate the impact of the International Nosocomial Infection Control Consortium (INICC) multidimensional approach on the reduction of ventilator-associated pneumonia (VAP) in adult patients hospitalized in 11 intensive care units (ICUs), from 10 hospitals, members of the INICC, in 10 cities of Turkey. METHODS: A prospective active before-after surveillance study was conducted to determine the effect of the INICC multidimensional approach in the VAP rate. The study was divided into two phases. In phase 1, active prospective surveillance of VAP was conducted using the definitions of the Centers for Disease Control and Prevention National Health Safety Network, and the INICC methods. In phase 2, we implemented the multidimensional approach for VAP. The INICC multidimensional approach included the following measures: (1) bundle of infection control interventions, (2) education, (3) outcome surveillance, (4) process surveillance, (5) feedback of VAP rates, and (6) performance feedback of infection control practices. We compared the rates of VAP obtained in each phase. A time series analysis was performed to assess the impact of our approach. RESULTS: In phase 1, we recorded 2,376 mechanical ventilator (MV)-days, and in phase 2, after implementing the multidimensional approach, we recorded 28,181 MV-days. The rate of VAP was 31.14 per 1,000 MV-days during phase 1, and 16.82 per 1,000 MV-days during phase 2, amounting to a 46 % VAP rate reduction (RR, 0.54; 95 % CI, 0.42-0.7; P value, 0.0001.) CONCLUSIONS: The INICC multidimensional approach was associated with a significant reduction in the VAP rate in these adult ICUs of Turkey.

Postoperative repeated respiratory insufficiency and thyrotoxicosis in molar pregnancy.

Following the removal of a hydatiform mole in a 34-year-old, 14-week pregnant patient, thyrotoxicosis and respiratory insufficiency attacks were twice unexpectedly repeated. The symptoms were resolved with the administration of plasmapheresis, antithyroid and ß-blocker drugs and non-invasive mechanical ventilation; however, she was again operated due to prolonged elevated ß-hCG.

The efficacy of supervised physiotherapy for the treatment of adhesive capsulitis.

OBJECTIVES: The aim of this study was to evaluate the efficacy of supervised physiotherapy supplemented with non-steroidal antiinflammatory drugs (NSAIDs) for treating adhesive capsulitis. BACKGROUND: Even though adhesive capsulitis is a common disorder, literature on its treatment is still limited. METHODS: Forty-five patients with adhesive capsulitis treated between 2004 and 2007 were reviewed retrospectively and 29 patients were enrolled in the study. All patients received a supervised physiotherapy program supplemented with NSAIDs. The outcome variables were determined as passive range of motion (ROM) values and intensity of shoulder pain. The clinical assessment was performed at the initial visit and immediately after physiotherapy. One to four years after the supervised physiotherapy, the resolved patients were reevaluated. RESULTS: The mean age of patients was 55.4+/-9.2 years. Fifteen patients (51.7 %) were women and 14 (48.3 %) were men. The average length of supervised physiotherapy was 3.5+/-0.5 weeks. After physiotherapy, the shoulder flexion, abduction and external rotation significantly increased (p<0.001) compared with the initial values. Additionally, shoulder pain significantly decreased (p<0.001). In the follow-up, 25 (86.2 %) of 29 patients were resolved with physiotherapy, whereas 4 (13.8 %) patients required capsular release operation because of residual functional impairment. As to long-term results of patients resolved with physiotherapy, 45.4 % of cases were cured with less than a 10 % loss in total passive ROM of the shoulder, 22.7 % of cases with a total ROM loss between 10 and 20 %, 18.1 % of cases with a total ROM loss from 20 to 30 %, and lastly, 13.6% of cases with a total ROM loss up to 40 %. CONCLUSIONS: Supervised physiotherapy supplemented with NSAIDs improves the ROM values in most of patients with adhesive capsulitis. A small percentage of patients may require operative treatment (Tab. 2, Fig. 1, Ref. 23). Full Text in free PDF www.bmj.sk.

The comparison of ultrasound treatment and local steroid injection plus splinting in the carpal tunnel syndrome: a randomized controlled trial.

OBJECTIVES: This study aimed to compare the efficacy of ultrasound treatment to local corticosteroid injection plus splinting in carpal tunnel syndrome (CTS). There is still limited knowledge on the efficacy of conservative treatment options for CTS. METHODS: Fourty-nine hands of 34 patients with CTS were enrolled in this study. Patients were randomly assigned to the ultrasound treatment (group A) or local corticosteroid injection plus splinting (group B). The primary outcome measures included clinical parameters such as symptoms severity score, visual analogue scale (VAS) pain, functional status score, grip strength and two-point discrimination. The secondary outcome measures were the electrophysiological tests. The examinations were performed at baseline, and then at 4th and 8th weeks. RESULTS: At the end of the study, a statistically significant improvement was obtained in all clinical parameters in the group B: VAS pain, severity of symptoms, functional status, grip strength (p < 0.001 for each) and two-point discrimination (p < 0.016). Also the group A showed significant improvements in all clinical parameters (p < 0.001 for each), except for the grip strength. Additionally, significant improvements in the median nerve sensory conduction velocity and distal motor latency were also found in both groups at the end of the 8 week follow-up period. There was no significant difference between the groups in the primary and secondary outcome measures, except for the grip strength. CONCLUSIONS: Both ultrasound treatment and corticosteroid injection plus splinting were effective on the clinical symptoms and the electrophysiological findings of CTS. Thus, the ultrasound therapy may be an alternative treatment for CTS, particularly in patient who do not accept injection or splinting (Tab. 3, Fig. 2, Ref. 36).

Serum vitamin D status and bone mineral density in fibromyalgia.

OBJECTIVES: To compare serum vitamin D levels and bone mineral density (BMD) values in patients with fibromyalgia and healthy controls. BACKGROUND: The so far available reports of low levels of vitamin D and low BMD values in patients with fibromyalgia are inconsistent. METHODS: Serum 25-hydroxyvitamin D (25-OHD) levels and BMD values were measured in thirty women with fibromyalgia and compared with thirty age-matched healthy women. Serum calcium, phosphorus, alkaline phosphatase and parathyroid hormone (PTH) levels were also measured. All participants completed the Fibromyalgia Impact Questionnaire (FIQ) and Hospital Anxiety and Depression Score (HADS). Pain severity was assessed with visual analog scale (VAS). RESULTS: Mean serum 25-OHD levels did not differ between the groups (fibromyalgia 10.57 +/- 10.46, controls 10.87 +/- 5.52 ng/l; p=0.89); nor did the frequency of vitamin D deficiency (25-OHD < or = 20 ng/l) in each group (fibromyalgia 86.7%, controls 96.7%; p=0.353). Although, mean serum PTH level was found significantly higher in fibromyalgic patients than in controls (p=0.014), only one patient and two of controls had barely elevated PTH levels. There was no relationship between vitamin D level and FIQ score (p=0.707) or HADS (p=0.824) or pain VAS (p=0.414). BMD values in the patients with fibromyalgia were comparable to those in controls at both, the lumbar spine (p=0.866) and femur neck (p=0.61). CONCLUSION: Neither vitamin D levels nor BMD values are different between women with and without fibromyalgia. In this cross-sectional study, mean serum PTH level was found higher in the fibromyalgic patients than in controls. Nevertheless, in order to confirm the findings of this preliminary study it is still necessary to perform a controlled longitudinal study (Tab. 2, Fig. 2, Ref. 35). Full Text in free PDF www.bmj.sk.

Potassium induced dilation in bovine coronary artery involves both inward rectifier potassium channels and Na+ /K+ ATPase.

Increases in extracellular potassium (K+) concentration (up to 20 mM) cause dilation in some blood vessels. This may be particularly important in myocardial ischemia because in this condition K+ is released from ischemic cells. In this study, we investigated mechanisms of effect of increased K+ concentration on the tone of isolated bovine coronary artery. Bovine coronary arteries were isolated and mounted in organ baths for isometric tension recording. After an equilibration period, arteries were contracted with serotonin (1 microM). When serotonin contraction reached a steady-state, K+ concentration of organ baths was increased from physiological levels to 10 mM, 14 mM, 18 mM or 22 mM in four groups of the arteries. After a washout period, this procedure was repeated in presence of ouabain, a blocker of Na+ /K+ ATPase or a K+ channel blocker (tetraethylammonium, 4-aminopyridine, glibenclamide or barium). Increasing K+ concentration of the organ baths to 10 mM, 14 mM and 18 mM caused dilation in the arteries. Ouabain abolished the dilation and barium (a blocker of inward rectifier K + channels) inhibited the dilation significantly.According to our results there is K+ -induced dilation in bovine coronary artery and it involves activation of both Na+ /K+ ATPase and inward rectifier K+ channels.

A new value of proximal femur geometry to evaluate hip fracture risk: true moment arm.

This study was undertaken to determine the influence of proximal femur geometry on hip fracture risk independent of bone mineral density. We examined 34 hip fracture subjects (17 men, 17 women) and 36 control subjects (18 men, 18 women). The control subjects were matched with the hip fracture patients by femoral neck bone mineral density (+ or - 0.100 g/cm(2)). Hip axis length (HAL), femoral axis length (FAL), femoral neck-shaft angle (Theta angle), lateral and medial femoral cortical thickness were measured on standart pelvic radiographs. In the literature, there are conflicting views of the relationship between femur geometry and hip fracture risk which may be explained by different definitions of some parameters. We investigated the effect of a new parameter called true moment arm (TMA) on hip fracture risk. Longer TMA may be correlated to higher transmission of impact energy to the femoral neck. Thus it may be useful to define fracture prone individuals. The results of this study showed that HAL, FAL and TMA were significantly longer in the hip fracture subjects compared to the control group (p<0.001). Hip fracture patients had thinner lateral and medial femoral cortical thickness (p<0.001). Theta angle was wider in the hip fracture group than in the control group (p<0.001). In conclusion, our study showed that evaluation of TMA in addition to HAL, FAL, Theta angle, MCT and LCT can be used to determine of the fracture risk independently of BMD.

Handwashing: a simple, economical and effective method for preventing nosocomial infections in intensive care units.

As most nosocomial infections are thought to be transmitted by the hands of healthcare workers, handwashing is considered to be the single most important intervention to prevent nosocomial infections. However, studies have shown that handwashing practices are poor, especially among medical personnel. This review gives an overview of handwashing in health care and in the community, including some aspects that have attracted little attention, such as hand drying and cultural issues determining hand hygiene behaviour. Hand hygiene is the most effective measure for interrupting the transmission of micro-organisms which cause infection, both in the community and in the healthcare setting. Using hand hygiene as a sole measure to reduce infection is unlikely to be successful when other factors in infection control, such as environmental hygiene, crowding, staffing levels and education, are inadequate. Hand hygiene must be part of an integrated approach to infection control. Compliance with hand hygiene recommendations is poor worldwide. While the techniques involved in hand hygiene are simple, the complex interdependence of factors that determine hand hygiene behaviour makes the study of hand hygiene complex. It is now recognized that improving compliance with hand hygiene recommendations depends on altering human behaviour. Input from behavioural and social sciences is essential when designing studies to investigate compliance. Interventions to increase compliance with hand hygiene practices must be appropriate for different cultural and social needs.

Pulmonary involvement in rheumatoid arthritis.

The primary objective of this investigation was to assess the relationships between clinical characteristics, lung involvement, and frequency of pulmonary involvement in rheumatoid arthritis (RA). Using high-resolution computed tomography (HRCT) and pulmonary function tests (PFT), we prospectively evaluated 52 patients with RA (eight males and 44 females, mean age 53.6 years). The HRCT was abnormal in 35 patients (67.3%), the most frequent abnormalities being reticulonodular patterns, which were found in 22 patients (62.9%), ground-glass attenuation (20%), and bronchiectasis (17%). In this group of patients, PFT results were normal in 13 patients (37%). Titers of rheumatoid factor and erythrocyte sedimentation rate were significantly higher in abnormal HRCT presence. Higher Larsen's score, advanced age, and severe disease were significant risk factors for lung involvement (p<0.001, p<0.01, and p<0.01, respectively) and are suggested by our data to be statistically significant predictors of lung involvement in RA.

Pulmonary abscess due to leflunomide use in rheumatoid arthritis: a case report.

A 43-year-old woman had rheumatoid arthritis (RA) for 5 years and complained of fever, arthralgia/myalgia, and night sweating for a month. She had been receiving only leflunomide (20 mg/day) for 5 months. On admission, there was no evidence of active arthritis or vasculitic lesion. Laboratory evaluation showed an erythrocyte sedimentation rate of 145 mm/h and C-reactive protein of 160 mg/dl. All cultures were negative. Chest radiograph and computed tomography (CT) revealed a pulmonary abscess. Staphylococcus aureus multiplied in the culture of a purulent sample obtained from the abscess under ultrasonography. The leflunomide was stopped, and sultamicillin (IV 4x2 g/day) was started for a further 6 weeks. Four weeks later, the patient had completely recovered and CT showed significant improvement of the pulmonary abscess. Ten milligrams/day of prednisolone and 7.5 mg/week of methotrexate were started for RA treatment. The patient has been under control for 5 months without any further abscess or RA activation.

Effects of magnesium sulfate on tissue lactate and malondialdehyde levels after cerebral ischemia.

In the present study, the effects of magnesium sulfate on tissue lactate and malondialdehyde (MDA) levels after cerebral ischemia in rabbits were studied. The rabbits were divided equally into three groups. Group 1 (n = 8) was the sham-operated control group, in group 2 (n = 8) only cerebral ischemia was induced by clamping bilaterally the common carotid arteries for 60 min, and in group 3 (n = 8) magnesium sulfate was administered at a dose of 100 mg/kg i.v. within 5 min after opening the clamps. In group 1 EEG recordings were obtained immediately and 60 and 120 min after craniectomy. In groups 2 and 3 EEG recordings were obtained immediately after craniectomy but before clamping and 60 min after clamping. One hour after opening the clamps and taking EEG recordings, brain cortices were resected, and the concentrations of lactate and MDA were determined using spectrophotometric/enzymatic and thiobarbituric acid methods, respectively. In all groups, there were significant differences between MDA and lactate levels (p < 0.05). There were no significant differences in lactate levels between groups 2 and 3 (p > 0.05), and also the preischemic EEG grades were the same in all groups. Preischemic and postischemic EEG values were significantly different (p < 0.05), and there were also significant differences between postischemic EEG grades in groups 2 and 3 (p < 0.05). There was a correlation between postischemic EEG grades and MDA and lactate levels. These results demonstrate that cerebral ischemia-reperfusion injury leads to an increase in brain tissue lactate and MDA levels, that magnesium sulfate suppresses the increase of MDA and lactate concentrations, and that magnesium sulfate treatment improves the EEG changes. The EEG grades correlated well with MDA and lactate levels.

Warming and response to contractile agents in calf cardiac vein: role of the nitric oxide.

The effects of warming on the response to various contractile agents of calf cardiac vein were studied using 2.5-mm long cylindrical segments. Concentration-response curves for carbachol (10(-9)-3 x 10(-4) m), 5-hydroxytryptamine (5-HT; 10(-8)-3 x 10(-3)), potassium chloride (KCl; 10(-4)-5 x 10(-2) m) and calcium chloride (CaCl2; 10(-4)-10(-2)) were isometrically recorded at 37 and 41 degrees C (warming). During warming the sensitivity, but not the maximal response, of carbachol 5-HT, KCl, and CaCl2 was significantly higher than at 37 degrees C. Warming to 41 degrees C after treatment with NG-nitro-L arginine methyl esther (10(-5) m) did not modify the effect of warming. These results suggest that nitric oxide seems to have no role in the warming-induced responses in calf cardiac vein.

Non-cardiogenic pulmonary oedema in the course of verapamil intoxication.

Only three cases of non-cardiogenic pulmonary oedema secondary to high dose verapamil ingestion have been described in the literature. This report describes two girls, who developed pulmonary oedema in the course of massive dose verapamil ingestion (total of 6000 mg and 7200 mg). Left ventricule size and function was normal on transthoracic echocardiograms in both of these patients. They were treated successfully with mechanical ventilatory support. The report emphasises that this fatal complication may be seen with verapamil overdose and underlying mechanisms and therapeutic approach are discussed.

The role of K+ ions on the response to carbachol of calf coronary artery and cardiac vein during cooling.

The role of K+ ions on the vasoconstrictions induced by carbachol during cooling (28 degrees C) in the endothelium of a denuded calf coronary artery and cardiac vein (noncutaneous vessel) was studied. Carbachol (10(-9) - 3 x 10(-4) M) induced concentration-dependent contractions at both 37 degrees C and 28 degrees C. The sensitivity, but not the maximal response, of carbachol (10(-9) -3 x 10(-4) M) was significantly lower at 28 degrees C than at 37 degrees C. Cooling to 28 degrees C after treatment with tetraethylammonium (TEA, 10(-3) M) or ouabain (10(-5) M), after incubation in K+-free medium increased the sensitivity to carbachol in both preparations. The results suggest a role for K+ ions in the cooling-induced changes of noncutaneous vessels.