Long non-coding RNA 01126 promotes periodontitis pathogenesis of human periodontal ligament cells via miR-518a-5p/HIF-1α/MAPK pathway.
Periodontitis is a prevalent oral inflammatory disease, which can cause periodontal ligament to a local hypoxia environment. However, the mechanism of hypoxia associated long non-coding RNAs (lncRNAs) involved in periodontitis is still largely unknown. Microarray was performed to detect the expression patterns of lncRNAs in 3 pairs of gingival tissues from patients with periodontitis and healthy controls. The expression of lncRNA 01126 (LINC01126), miR-518a-5p and hypoxia-inducible factor-1α (HIF-1α) in periodontal tissues and in human periodontal ligament cells (hPDLCs) under hypoxia was measured by quantitative real-time polymerase chain reaction or western blot. Fluorescence in situ hybridization and cell fraction assay were performed to determine the subcellular localization of LINC01126 and miR-518a-5p. Overexpression or knockdown of LINC01126 or HIF-1α was used to confirm their biological roles in hPDLCs. MTT assays were performed to evaluate hPDLCs proliferation ability. Flow cytometry was used to detect apoptosis. ELISA was used to measure the expression levels of interleukin (IL)-1β, IL-6, IL-8 and TNF-α. Dual-luciferase reporter assays were performed to assess the binding of miR-518a-5p to LINC01126 and HIF-1α. RNA immunoprecipitation assay was used to identify whether LINC01126 and miR-518a-5p were significantly enriched in AGO-containing micro-ribonucleoprotein complexes. We selected LINC01126, which was the most highly expressed lncRNA, to further verify its functions in periodontitis-induced hypoxia. The expression of LINC01126 was increased in periodontal tissues. In vitro experiment demonstrated that LINC01126 suppressed proliferation, promoted apoptosis and inflammation of hPDLCs under hypoxia via sponging miR-518a-5p. Moreover, we identified HIF-1α acted as a direct target of miR-518a-5p in hPDLCs and LINC01126 promoted periodontitis pathogenesis by regulating the miR-518a-5p/HIF-1α/MAPK pathway. LINC01126 promotes periodontitis pathogenesis of hPDLCs via miR-518a-5p/HIF-1α/MAPK pathway, providing a possible clue for LINC01126-based periodontal therapeutic approaches.


Producing protein microarrays from DNA microarrays.
The development of protein microarrays makes possible interaction-based protein assays in miniaturised, multiplexed formats. A major requirement determining their uptake and use is the availability and stability of purified, functional proteins for immobilisation. With conventional methods, involving individual expression and purification of recombinant proteins, the cost of commercial high-content protein arrays is often found to be prohibitively high. Moreover, due to the need for specialised microarray production equipment, custom-made protein arrays containing more focussed sets of proteins of interest are also in little use. In the DNA array to protein array technology described herein, repeated economical printing of protein microarrays from a reusable template DNA microarray is performed on demand by cell-free -protein synthesis. Once the template DNA microarray has been obtained, protein microarrays are made using purely macro-handling procedures, making protein arraying accessible without sophisticated microarraying apparatus.


Perfluorooctanoic acid-induced immunomodulation in adult C57BL/6J or C57BL/6N female mice.
Perfluorooctanoic acid (PFOA), an environmentally persistent compound of regulatory concern, has been reported to reduce antibody responses in mice at a single dose. The aim of this study was to evaluate PFOA effects on humoral and cellular immunity using standard assays for assessing immune function, and to derive dose-response data. C57BL/6J mice received 0 or 30 mg PFOA/kg/day for 10 days; half of the exposed groups were switched to vehicle and half continued on PFOA for five days. C57BL/6N mice received 0-30 mg/kg/day of PFOA in drinking water for 15 days. Mice were immunized with sheep red blood cells or sensitized to bovine serum albumin in Freund's complete adjuvant on day 10 of exposure; immune responses were determined 1 day post-exposure. We found that 30 mg PFOA/kg/day given for 10 or 15 days reduced IgM synthesis; serum collected 1 day postexposure contained 8.4 x 10(4) or 2.7 x 10(5) ng PFOA/mL, respectively. IgM synthesis was suppressed at exposures > or = 3.75 mg PFOA/kg/day in a dose-dependent manner, and IgG titers were elevated at 3.75 and 7.5 mg PFOA/kg/day. Serum PFOA at 3.75 mg/kg/day was 7.4 x 10(4) ng/mL 1 day postexposure, or 150-fold greater than the levels reported in individuals living near a PFOA production site. Using a second-degree polynomial model, we calculated a benchmark dose of 3 mg/kg/day, with a lower bound (95% confidence limit) of 1.75 mg/kg/day. Cell-mediated function was not affected. IgM antibodies were suppressed after PFOA exposure. The margin of exposure for reduced IgM antibody synthesis was approximately 150 for highly exposed human populations.


Patients without records and records without patients: review of patient records in primary care and implications for surveillance of antibiotic prescribing in rural China.
We need to monitor patterns of antibiotic prescribing in order to develop and evaluate antibiotic stewardship interventions in rural China. As part of a multidisciplinary study of antibiotic use in Anhui Province we assessed the validity of electronic records (e-records) as a source of surveillance data. One township healthcare centre and one village clinic were selected in each of three different counties. Patients with symptoms of Upper Respiratory Tract Infection (URTI), exacerbation of Chronic Obstructive Pulmonary Disease (COPD) or Urinary Tract Infection (UTI) were recruited consecutively. Researchers observed and documented clinic consultations and interviewed each of the study participants. E-records were compared to clinic observations and patient interviews. A total of 1030 patients were observed in clinic. Antibiotics were prescribed in 917 (89%) of consultations. E-records were created only for individuals with health insurance, with considerable between-site variation in completeness (0 to 98.7% of clinic consultations) and in the timing of documentation (within-consultation up to weeks afterwards). E-record accuracy was better in relation to antibiotics (82.8% of e-records accurately recorded what was prescribed in clinic) than for diagnosis and symptoms (45.0 and 1.1% accuracy). Only 31 participants (3.0%) presented with UTI symptoms. We have confirmed very high rates of outpatient antibiotic prescribing in rural Anhui province. E-records could provide useful information to inform stewardship interventions, however they may be inaccurate and/or biased. Public Health authorities should focus on improving technical infrastructure and record-keeping culture in outpatient settings. Further research is needed into community treatment of UTIs.


Interaction between pH-shifted ovalbumin and insoluble neohesperidin: Experimental and binding mechanism studies.
In this study, ovalbumin (OVA) formed a complex with neohesperidin (NH) via a pH-shifting method. The NH-OVA complex self-assembled into NH-OVA nano-particles, which were then characterized and whose binding mechanism was evaluated by using multi-spectroscopic, thermodynamics, and molecular docking simulation methods. Fluorescence intensity decreased after OVA was complexed with NH. The binding constant of the OVA-NH complex was in the order of 6.32 × 105 M-1 suggesting that the complex is stable. Circular dichroism (CD) analysis showed that α -helix content increased, β-folding, β -turning, and irregular crimp content decreased after OVA and NH binding. Isothermal titration calorimetry results showed that hydrophobic interactions and hydrogen bonds made an important impact in the complex formation. The molecular docking results revealed that Van der Waals forces and hydrogen bonds contributed to the free binding energy of the complex. There were multiple possible surface binding sites between OVA with NH. The obtained results provide new insights into the interaction mechanism of OVA and NH, and as a vehicle for NH, the OVA has shown promising applications in functional foods.


Clinician assessment of blood alcohol levels among emergency department patients.
Alcohol intoxication is a significant public health concern and is commonly seen among emergency department (ED) patients. This study was undertaken to identify the accuracy of clinician assessment of blood alcohol levels among emergency department patients. This prospective survey study was conducted at a Level 1 Trauma Center. Eligible study participants included physicians, nurses, and medical students involved in the care of patients who had BAC. Clinicians estimated the BAC prior to results availability. Among 243 clinicians, the mean difference between the estimated BAC and actual BAC was 17.4 (95% CI: 4.7 to 30.1). Providers tended to overestimate the actual BAC level. The accuracy between roles (attendings, residents, RNs, students) was not significant (ANOVA p-value 0.90). Accuracy was not correlated with age of the patient (Pearson correlation 0.04, p-value 0.54). Accuracy was not associated with the patient's gender (Student's t-test two-tailed p-value 0.90), ethnicity (White versus all others, t-test p-value 0.31), nor insurance (government versus not government, t-test p-value 0.81). The average accuracy value was associated with mode of arrival (t-test p-value 0.003). The average accuracy for walk-in subjects was -14.9 (CI: -32.8 to 3.1) compared to ambulance arrivals 28.3 (CI: 12.7 to 44.0). Providers underestimated BAC for walk-ins and overestimated BAC for ambulance arrivals. Among 107 patients with a BAC of 0, clinician estimates ranged from 0 to 350. Clinicians estimated non-zero BAC levels in 17% of patients with BAC of 0 (N = 18). Clinicians' estimates of BAC were often inaccurate, and often overestimated the BAC.


The role of histidine in human epidermis.
Incorporation of radioactively-labelled histidine into half-thickness skin and into the major protein fraction extracted from epidermis by urea (protein Fraction A) has been measured and compared with the amount of labelled urocanic acid appearing in the same specimen. After a short lag period, incorporation into half-thickness skin and into protein Fraction A was found to be linear up to 23 h of the study. Histidine appeared to be initially preferentially deaminated to urocanic acid. The somewhat erratic presence of urocanic acid at later periods is attributed to its high instability and solubility.


Transfection-free and scalable recombinant AAV vector production using HSV/AAV hybrids.
Adeno-associated virus (AAV) vectors are highly efficient tools for use in gene therapy. Current production methods rely on plasmid transfection and are not generally considered amenable to scale-up. To improve recombinant AAV (rAAV) vector production in terms of both final titre and simplicity, we constructed recombinant herpes simplex virus (HSV) vectors, either disabled (ICP27 deleted) or nondisabled, encoding the AAV rep and cap genes. We also integrated an rAAVGFP construct into the nondisabled vector and also into a second pair of HSV vectors (disabled and nondisabled) not expressing rep and cap. Transgene incorporation and expression was confirmed by Southern and Western blot, respectively. Optimal double-infection ratios were established for disabled and nondisabled pairs of rep/cap-expressing and rAAVGFP-containing vectors, resulting in up to 1.55 x 10(12) rAAV capsids and 4 x 10(8) expression units from approximately 1 x 10(7) BHK producer cells. Functionality of the prepared vector was confirmed by the detection of abundant green fluorescent protein (GFP) expression following injections of rAAV preparations into the rat brain. This paper therefore describes a simple, efficient, and transfection-free rAAV production process based on the use of HSV and not relying on a proviral cell line that, with appropriate scale-up, could yield quantities of rAAV sufficient for routine clinical use.


Draft Genome Sequences for Bacteria Associated with Root Nodules of Alnus incana in New England.
Nine bacterial strains isolated from the root nodules of Alnus incana were sequenced to determine their potential roles in plant health. The selected bacterial isolates belonged to the genera Bacillus, Herbaspirillum, Pantoea, Paenibacillus, and Rothia. Here, we report the draft genome sequences.


MAIA microperimeter for short-duration fixation stability measurements in central vision loss: Repeatability and comparison with the Nidek MP1.
This study reports the repeatability of 20 s-duration fixation stability measurements recorded with the Macular Integrity Assessment (MAIA) microperimeter in patients with central vision loss, in contrast to the Nidek MP1 microperimeter. Fixation stability was recorded in 39 eyes of 25 patients with macular disease using MAIA and the MP1 for 20 s intervals, twice for each eye, with each instrument. Twenty eyes were identified as the better eye (BE) and 19 eyes as the worse eye (WE). Fixation stability was quantified with the 95% bivariate contour ellipse area (BCEA), logarithmically transformed. Bland-Altman plots were used to determine the 95% limits of agreement. For MAIA, the 95% limits of agreement were ±0.84 log deg2 for the BE and ±0.66 log deg2 for the WE. Similarly, for the MP1 these limits were ±0.48 log deg2 for the BE and ±0.72 log deg2 for the WE. Inter-device repeatability was modest, ±1.09 log deg2 for the BE and ±1.01 log deg2 for the WE, and a proportional bias was detected. Occasionally, MAIA did not register all the expected number of data points, and included far outliers in the BCEA calculation; the inter-device repeatability did not improve when these outliers were removed. Repeatability of 20 s-duration fixation stability examination in patients with central vision loss is specific to the instrument used. We recommend that only data from same type of microperimeter with the same fixation duration should be compared when using fixation stability as an outcome measure to monitor disease progression, effect of treatment or in clinical trials.


Binding of [3H]forskolin to solubilized preparations of adenylate cyclase.
The binding of [3H]forskolin to proteins solubilized from bovine brain membranes was studied by precipitating proteins with polyethylene glycol and separating [3H]forskolin bound to protein from free [3H]forskolin by rapid filtration. The Kd for [3H]forskolin binding to solubilized proteins was 14 nM which was similar to that for [3H]forskolin binding sites in membranes from rat brain and human platelets. Forskolin analogs competed for [3H]forskolin binding sites with the same rank potency in both brain membranes and in proteins solubilized from brain membranes. [3H]forskolin bound to proteins solubilized from membranes with a Bmax of 38 fmol/mg protein which increased to 94 fmol/mg protein when GppNHp was included in the binding assay. In contrast, GppNHp had no effect on [3H]forskolin binding to proteins solubilized from membranes preactivated with GppNHp. Solubilized adenylate cyclase from non-preactivated membranes had a basal activity of 130 pmol/mg/min which was increased 7-fold by GppNHp. In contrast, adenylate cyclase from preactivated membranes had a basal activity of 850 pmol/mg/min which was not stimulated by GppNHp or forskolin. Thus, the number of high affinity binding sites for [3H]forskolin in solubilized preparations correlated with the activation of adenylate cyclase by GppNHp via the guanine nucleotide binding protein (GS).


Maternal discussions about pregnancy and adolescents, attitudes toward pregnancy.
To identify aspects of maternal-adolescent relationships that are associated with high ambivalence toward pregnancy. This was a cross-sectional study of 350 inner-city, African-American females between the ages of 14 and 17 years residing in the Philadelphia metropolitan area. Adolescent females and their resident female caretaker (usually the biological mother) were both interviewed. Multiple regression analyses were used to predict adolescents' pregnancy attitudes from maternal-adolescent discussions about pregnancy, perceived maternal disapproval of pregnancy, and relationship satisfaction. Greater levels of discussion about negative consequences of pregnancy, stronger levels of perceived maternal disapproval, and higher levels of satisfaction all were associated with more negative attitudes towards pregnancy on the part of the adolescent. Maternal communication about the negative consequences of experiencing a pregnancy and the quality of the mother-daughter relationship more generally may impact on adolescent attitudes toward pregnancy.


Drain fixation made foolproof.
A reliable method of securing drain tubing is presented. Knots made from two half-hitches have been used in the sailing world for many years. They provide a strong, slip-free knot which is simple to learn, tie, teach and easy to remove.


Cardiovascular response to increasing theophylline concentrations.
The circulatory effects of plateau plasma theophylline concentrations of 5, 10 and 20 mg/l were examined in normal male volunteers using forearm plethysmography, systolic time intervals and echocardiography. Systemic arterial and forearm venous pressures were not significantly altered during theophylline infusions. Forearm blood flow increased an average 9.4% at 5 mg/l, 15.3% at 10 mg/l and 41.2% at 20 mg/l. Forearm venous distensibility was maximally increased by 20.9% at 10 mg/l. Changes in heart rate and systolic time intervals were apparent only at concentrations between 10-20 mg/l. Heart rates increased from 3-16 beats/min. The left ventricular ejection time index and isovolumetric contraction time were significantly reduced with an increase in the PEP/LVET ratio consistent with a reduction in cardiac pre-load and an increase in inotropy. However, the interindividual variability in circulatory responses was great and consistent alterations in cardiac stroke volume and ejection fraction were not observed in either the supine or 60 degrees passive-tilt positions. The variable responses cannot be due to variations in drug disposition between subjects since the range of plasma theophylline values at each concentration plateau was narrow.


Maternal floor infarction of the placenta: association with central nervous system injury and adverse neurodevelopmental outcome.
To compare cranial ultrasound studies and neurodevelopmental outcome of preterm infants affected by maternal floor infarction (MFI) of the placenta to gestational age-matched controls over an 8-year period from a single institution. Retrospective case/control study. Compared to gestational age-matched controls, infants born to mothers with MFI had a higher incidence of CNS injury on neonatal cranial ultrasound examinations and at follow-up were more likely to have a suspicious or abnormal neurologic examination. MFI cases had lower developmental scores in all areas tested and were more likely to have neurodevelopmental impairment. Infants born to mothers with MFI should have serial neonatal cranial ultrasound examinations to detect CNS injury and neurodevelopmental assessment during early childhood.


An improved DNA extraction method for detecting Bacillus subtilis spores in spiked foods and beverages.
Bacillus and Paenibacillus spp. are essential aerobic spoilage bacteria in various food industry sectors. Spoilage from microorganisms occurs at many points throughout food production systems. Due to their complex wall structures, spores can resist heat, radiation, chemical agents, and enzymatic treatments. An alkaline lysis and mechanical disruption combination method was developed and evaluated to counter this. This combination method effectively improved DNA extraction from B. subtilis spore cells spiked into food (solid) and beverages (liquid milk and coffee) at concentrations down to 102 CFU/mL or g when spiked into food matrices and drinks. Released DNA recoveries were 27 % and 25 % for potato salad and 38 % and 36 % for whole corn spiked at 106 and 103 CFU/mL concentrations. Conversely, there was a low recovery for wheat flour (10 % and 8.8 %) and milk powders (12 % and 25 %) at 106 and 103 CFU/mL spiked concentrations. The combination method provides rapid, specific, reliable, and accurate signature sequences identification for the detection and presence confirmation of psychrophilic and psychrotolerant spoilage spore cells, improving food spoilage assessments and food control applications.


Dynamic Radiographic Results of Different Semi-Rigid Fusion Devices for Degenerative Lumbar Spondylolisthesis: "Dynamic Rod" vs. "Dynamic Screw Head".
To assess the clinical outcomes and compare the segmental range of motion (ROM) at the implanted L4-L5 level, the cranial and caudal adjacent levels, and the ROM of the whole lumbar spine after semi-rigid lumbar fusion with 2 different devices. Patients with neurogenic claudication, due to grade 1 spondylolisthesis and spinal stenosis at levels L4-L5, were treated either Bio-flex® (n = 28) and Cosmic® (n = 23); discectomy was not performed at any level. The clinical outcomes were compared between the 2 groups. All patients underwent neutral, flexion, and extension radiography before the surgery and after 2 years postoperatively. ROM was assessed at the level of L4-L5, L3-L4, L5-S1, and at the whole lumbar spine. According to clinical outcomes, 82% and 78% of patients in the BioFlex and Cosmic groups, respectively, had a good or excellent result. In both groups, there was significant reduction of the segmental ROM at the implanted L4-L5 level (p = 0.039 and 0.011). These outcomes may play a role in decreasing the risk of ASD after dynamic stabilization, at least 2 years after surgery.


Modeling myocardial growth and hypertrophy in engineered heart muscle.
The introduction of biomimetic culture paradigms has advanced myocardial tissue engineering fundamentally, enabling today the provision of engineered rodent and human heart muscle with features characteristically found in postnatal myocardium. This is in marked contrasts to "flat" cardiomyocyte cultures with their typically low degree of organotypic maturation. Here, we discuss the collagen hydrogel-based engineered heart muscle (EHM) technology and provide background information on its use in simulations of myocardial growth and disease.


Predicting Responses to Pregabalin for Painful Diabetic Peripheral Neuropathy Based on Trajectory-Focused Patient Profiles Derived from the First 4 Weeks of Treatment.
Prediction of final clinical outcomes based on early weeks of treatment can enable more effective patient care for chronic pain. Our goal was to predict, with at least 90% accuracy, 12- to 13-week outcomes for pregabalin-treated painful diabetic peripheral neuropathy (pDPN) patients based on 4 weeks of pain and pain-related sleep interference data. We utilized active treatment data from six placebo-controlled randomized controlled trials (n = 939) designed to evaluate efficacy of pregabalin for reducing pain in patients with pDPN. We implemented a three-step, trajectory-focused analytics approach based upon patient responses collected during the first 4 weeks using monotonicity, path length, frequency domain (FD), and k-nearest neighbor (kNN) methods. The first two steps were based on combinations of baseline pain, pain at 4 weeks, weekly monotonicity and path length during the first 4 weeks, and assignment of patients to one of four responder groups (based on presence/absence of 50% or 30% reduction from baseline pain at 4 and at 12/13 weeks). The third step included agreement between prediction of logistic regression of daily FD amplitudes and assignment made from kNN analyses. Step 1 correctly assigned 520/939 patients from the six studies to a responder group using a 3-metric combination approach based on unique assignment to a 50% responder group. Step 2 (applied to the remaining 419 patients) predicted an additional 121 patients, using a blend of 50% and 30% responder thresholds. Step 3 (using a combination of FD and kNN analyses) predicted 204 of the remaining 298 patients using the 50% responder threshold. Our approach correctly predicted 90.0% of all patients. By correctly predicting 12- to 13-week responder outcomes with 90% accuracy based on responses from the first month of treatment, we demonstrated the value of trajectory measures in predicting pDPN patient response to pregabalin. www.clinicaltrials.gov identifiers, NCT00156078/NCT00159679/NCT00143156/NCT00553475. Pfizer. Plain language summary available for this article.


The use of laminarias for osmotic dilation of the cervix in gynecological brachytherapy applications.
Osmotic dilators (laminarias) have been used for gradual nontraumatic dilation of the cervical canal for various intrauterine procedures; however, this technique has not been well accepted in gynecological brachytherapy. The purpose of this study was to evaluate the efficacy of osmotic cervical dilation for brachytherapy in gynecologic cancer patients, without the use of general/regional anesthesia, and to assess patient tolerance, complications, and outcome. Thirteen brachytherapy procedures were performed in 6 patients with clinical Stages I and II endometrial (5) and Stage IB cervical cancer (1), who were unable to tolerate general/regional anesthesia because of severe medical problems. An osmotic dilator (synthetic laminaria) was inserted into the cervical os 10-12 h before each brachytherapy procedure and removed just before the procedure. Standard Fletcher-Suit-Delclos tandem insertions with vaginal colpostats or cylinders were then performed. Degree of cervical dilation, patient discomfort, procedure time, intra- and postoperative complications were recorded, and local control and survival were assessed. Median follow-up was 31 months (range: 8-35 months). The diameter of the dilated cervical os after laminaria removal was adequate (> or = 5 mm) for tandem insertion, and no additional mechanical dilation was required in all but one procedure (1 of 13). All procedures were performed without general/regional anesthesia. The mean duration of the procedures was 44 min (range, 20-60 min). Discomfort was minimal in all cases. There were no intra- or postoperative complications. All patients maintained local control until death (1 of metastatic disease, 2 of intercurrent disease) or last follow-up (2 with no evidence of disease, 1 alive with metastatic disease). This preliminary study suggests that osmotic cervical dilation with a synthetic laminaria is a useful technique to facilitate intrauterine tandem insertion in patients who cannot tolerate general/regional anesthesia. This technique may reduce treatment-associated morbidity, shorten procedure time, and allow the delivery of adequate radiation therapy in this uncommon but challenging patient population.


Development and validation of an LC-MS/MS method for quantifying nine antimicrobials in human serum and its application to study the exposure of Chinese pregnant women to antimicrobials.
To study the prevalence of the exposure of pregnant women to antimicrobials, a sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine nine antimicrobials, namely sulfadimidine, sulfapyridine, sulfadiazine, sulfathiazole, ofloxacin, ciprofloxacin, norfloxacin, tetracycline, and lincomycin, in human serum. The sample preparation procedure included protein precipitation followed by a cleanup step with solid phase extraction (SPE). Separation was carried out using a CORTECS T3 column (100 × 2.1 mm, 2.7 µm) by gradient elution with a runtime of 8.0 min. Detection was performed on a triple quadruple tandem mass spectrometer with scheduled multiple reaction monitoring (sMRM) in positive ion scan mode. The calibration curves were linear over the concentration range of 0.5-50 ng/ml, and the limit of quantitation was between 0.01 and 0.2 ng/ml. For each level of quality control samples, the inter- and intra-assay precision values were less than 12.0%, and the accuracy ranged from 86.1% to 109.0%. No significant matrix effect or carryover was observed. The antimicrobials of interest were stable under all investigated conditions. The validated method was applied to analyze clinical samples from pregnant women in China, and 10 out of 500 samples showed the presence of antimicrobial residues. Moreover, compared with the time-resolved fluoro-immunoassay (TRFIA) method, the developed method showed greater sensitivity and specificity. This study provides a simple and rapid LC-MS/MS method for the simultaneous measurement of nine antimicrobials in serum samples, which could be a useful tool in clinical utilization.


Pharmacological analysis of analgesia and self-stimulation elicited by electrical stimulation of catecholamine nuclei in the rat brain.
The relationship between intracranial self-stimulation (ICSS) and stimulation-produced analgesia (SPA) was investigated in that rat employing an operant bar-press response and a modification of the hot-plate test. ICSS and SPA were elicited through bipolar electrodes chronically implanted in two catecholamine nuclei; the nucleus locus coeruleus (LC) and the substantia nigra (SN). These sites have previously been shown to yield both phenomena. SPA was shown to be of a magnitude similar to that of morphine. In addition, SPA deriving from both LC and SN was significantly reversed by the specific opiate antagonist, naloxone. The intensity of the stimulating current sufficient to induce SPA was found to be higher than that required for ICSS. The pharmacological susceptibilities of the two phenomena were tested by administering a number of drugs: haloperidol, propranolol, pimozide and AMPT attenuated ICSS in both LC and SN, while leaving SPA unaffected. In contrast, methysergide and PCPA blocked SPA and simultaneously facilitated ICSS. The present results indicate a dissociation of ICSS and SPA from LC and SN at both physiological and neurochemical levels; ICSS rates appeared to be a function of catecholaminergic tone, while SPA depended upon the integrity of serotonin transmission.


[Current methods of detection and localization of vagal chemodectomas].
The modern methods of diagnosis were analysed in 19 patients with chemodectoma of vagal glomus verified surgically and morphologically. The cytological investigation of the tumour punctates was made in 9 patients, ultrasonic introscopy in 2, computer tomography in 13 and arteriography in 17 patients. The informativeness of cytological and ultrasonic studies is low, whereas computer tomography and carotid arteriography are regarded as the method of choice for diagnosis and assessment of vagal chemodectoma prevalence.


Bioaccessibility of polycyclic aromatic compounds (PAHs, PCBs) and trace elements: Influencing factors and determination in a river sediment core.
Organic matter (OM), clays, sand or time are factors possibly influencing the bioaccessibility of polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs) from sediments. An experimental design was performed to monitor and quantify this process. The bioaccessible fraction, linked to the rapidly-desorbing fraction (Frap) of contaminants, was assessed through a non-exhaustive extraction using a carboxymethyl-β-cyclodextrin polymer. OM content was the most influential factor as regards Frap. Clay percentage was a slightly influential factor for PAHs while the interaction sand × OM was a slightly influential factor for PCBs. Frap was also determined in a sediment core collected from Martot's Pond (France). The higher the PAH/PCB concentration in this sediment, the higher the bioaccessible fraction. The relationship between a lower bioaccessibility and a higher number of PAHs cycles or PCB chlorines was linear. OM content impacted on Frap only for PAHs. Sequential extractions of some trace elements were also performed to evaluate their mobility. Cu, Cr, Pb, Ni were the less bioaccessible. A great part of As, Cd and Zn was found in the most bioaccessible sediment fractions. The 40-65 cm section might be considered as the most negatively impacting on the aquatic fauna, due to Cd and Zn high bioaccessible concentrations.


Structural stability of glycophorin. Effects of heat and guanidine . HCl.
The effects of guanidine hydrochloride and high temperature on human glycophorin and sialic acid-free glycophorin were monitored by circular dichroism, viscosity, and fluorescence of 1-anilino-8-naphthalane sulfonate (ANS). The following observations were made: 1. Glycophorin and its sialic acid-free counterpart are unusually stable to both guanidine . HCl and heat. 2. CD and viscosity measurements indicate that guanidine . HCl neither causes a cooperative unfolding nor generates a random coil. 3. The ANS binding site is much more sensitive to guanidine . HCl than the ellipticity at 220 nm (theta 220). 4. The effect of temperature on CD is reversible whereas the effect of guanidine . HCl is not. 5. The carbohydrate moiety influences the viscosity, and also contributes to the changes in theta 220 when solutions of glycophorin are heated. These unusual properties indicate a complex mechanism of unfolding for this structurally stable macromolecule.


Effects of semistarvation on transintestinal D-glucose transport and D-glucose uptake in brush border and basolateral membranes of rat enterocytes.
The present work shows that semistarvation (8-10 g of food for 10 days) increases net D-glucose, Na+ and water transport in the everted and perfused rat jejunum. A linear and positive correlation between cell sugar concentration and transport was found in control and semistarved rats, but the phenomenon was more relevant only in semistarved animals. Membrane vesicle experiments showed that semistarvation increases sugar overshoot only in brush border membrane vesicles, while this situation does not occur in basolateral membrane vesicles. The effect of partial food deprivation seems to enhance net sugar transport by increasing sugar entry across the apical membrane of enterocytes.


A comparison of 3-parameter, single-species population models, in relation to the management of brushtail possums in New Zealand.
A range of published 3-parameter, single-species models are compared using possums in New Zealand as an example. Most models yield similar population growth curves and similar estimates of maximum sustainable yield (MSY). A new model of competition for refuges, and interactive models of competition for food, both yield growth curves peaked to the right. In the second case, the relationship between the shape of the curve and the parameters of the interactive model is described. Three management consequences of asymmetric (rightward-peaked) growth as opposed to logistic growth are discussed. First, where the animal is a pest a greater control effort is required to reduce density. Second, for sustained yield harvesting the best policy is an adaptive one which initially requires no knowledge of the shape of the growth curve. Third, sustainable yields may be 50% higher than those predicted by the simple logistic model.


Competitive adsorption of phenolic compounds from aqueous solution using sludge-based activated carbon.
Preparation of activated carbon from sewage sludge is a promising approach to produce cheap and efficient adsorbent for pollutants removal as well as to dispose of sewage sludge. The first objective of this study was to investigate the physical and chemical properties (BET surface area, ash and elemental content, surface functional groups by Boehm titration and weight loss by thermogravimetric analysis) of the sludge-based activated carbon (SBAC) so as to give a basic understanding of its structure and to compare to those of two commercial activated carbons, PICA S23 and F22. The second and main objective was to evaluate the performance of SBAC for single and competitive adsorption of four substituted phenols (p-nitrophenol, p-chlorophenol, p-hydroxy benzoic acid and phenol) from their aqueous solutions. The results indicated that, despite moderate micropore and mesopore surface areas, SBAC had remarkable adsorption capacity for phenols, though less than PICA carbons. Uptake of the phenolic compound was found to be dependent on both the porosity and surface chemistry of the carbons. Furthermore, the electronegativity and the hydrophobicity of the adsorbate have significant influence on the adsorption capacity. The Langmuir and Freundlich models were used for the mathematical description of the adsorption equilibrium for single-solute isotherms. Moreover, the Langmuir-Freundlich model gave satisfactory results for describing multicomponent system isotherms. The capacity of the studied activated carbons to adsorb phenols from a multi-solute system was in the following order: p-nitrophenol > p-chlorophenol > PHBA > phenol.


Surgical treatment of tumours of the skull base.
To describe the special features of surgical removal of tumours of the skull base and to review a series of patients treated for skull base tumours. A retrospective review of patients followed up for between six months and six years. A unit specialising in surgery of the skull base at a tertiary referral centre. Sixty-six patients with skull base tumours were referred to the unit between February 1984 and December 1989. Fifty-six of the patients underwent complete or partial surgical removal of the tumour. Radiotherapy was used as primary treatment in 10 patients, for tumour recurrence in four patients and as adjuvant postoperative treatment in 10 patients. In the majority of patients the tumours were benign and have been controlled or are considered cured. Of the seven patients with malignant tumours five have died and two are long-term survivors (one has multiple metastases). Three patients with histologically "benign" tumours died of multiple metastases. There have been a total of 10 deaths in the series but only one occurred in the perioperative period. Cranial nerve palsy was the major postoperative complication. Modern microsurgical techniques and modern anaesthesia and intensive care skills lead to a cure in many patients with skull base tumours previously considered inoperable.


Cytisine basicity, solvation, logP, and logD theoretical determination as tool for bioavailability prediction.
Cytisine, an α4β2 nicotinic receptor partial agonist, is a plant alkaloid widely used as a smoking cessation agent. Despite long history of use, knowledge on pharmacokinetics of cytisine still demands an extension. This work is aimed at theoretical determination of physicochemical parameters that affect the bioavailability of cytisine. The acidic dissociation constant, Gibbs free energy of solvation in water and n-octanol as well as n-octanol/water partition coefficient and n-octanol/water distribution coefficient of cytisine were calculated as quantities corresponding to its solubility and permeability. Cytisine structure was optimized with several quantum chemical methods-ab initio: HF and MP2, and DFT functionals (B3LYP, B3LYP-D3, CAM-B3LYP, M06-2X, TPSS, VSXC) with 6-311++G(d,p) basis set. Solvation of cytisine in water and n-octanol was determined with the SMD continuum model. It was shown that lipophilicity of cytisine depends on the pH of an environment. Protonated cytisine, the most populated state under acidic conditions, is characterized by enhanced hydrophilicity. Then neutral cytisine, dominating in a basic environment, demonstrates more lipophilic character. It appears that cytisine is very well soluble in the gastrointestinal (GI) tract fluids. Then the distribution of cytisine ought to occur very rapidly. However, permeability of cytisine through the mucous membrane of the GI tract may be limited, leading to the diminished bioavailability.


Carotid Atherosclerotic Plaque Alters the Direction of Longitudinal Motion in the Artery Wall.
Longitudinal motion of the artery, a cyclical, bidirectional movement of the wall in the long axis of the artery, has recently gained interest in the characterization of artery function. The aim of this study was to evaluate longitudinal motion in patients with internal carotid atherosclerotic plaques. Speckle tracking ultrasound was used to assess common carotid artery wall motion in 12 patients with carotid plaque causing either moderate (50%-79%) or severe (80%-99%) stenosis based on the North American Carotid Endarterectomy Trial, and 23 healthy participants. Although healthy individuals were found to have a retrograde wall motion pattern, a distinct anterograde pattern was noted with plaque presence. Importantly, patients with severe plaque stenosis had greater anterograde motion (0.53 ± 0.36 mm) than those with moderate stenosis (0.17 ± 0.15 mm) (p < 0.05), likely owing to high wall shear stresses associated with greater peak systolic velocities at the site of stenosis (severe: 342.0 ± 99.4 cm/s, moderate: 177.5 ± 31.2 cm/s, p < 0.01). There were no differences in peak systolic velocities at plaque-free segments between plaque groups (severe: 80.2 ± 24.8 cm/s, moderate: 92.7 ± 23.0 cm/s). Blood flow at stenotic areas better predicted motion than plaque-free segments. We conclude that the presence of carotid plaque can have significant influence on longitudinal motion, with significantly greater anterograde displacements with increased stenosis. Future studies are needed to further investigate carotid artery wall mechanics.


Continuous monitoring of cerebrocortical blood flow during stimulation of the cerebellar fastigial nucleus: a study by laser-Doppler flowmetry.
Laser-Doppler flowmetry was used to continuously monitor cortical CBF during electrical stimulation of the fastigial nucleus (FN). Rats were anesthetized with isoflurane (0.75-5%), paralyzed, and artificially ventilated. The LDF probe was placed over a target region of the parietal cortex through a burr hole. The hypertension associated with FN stimulation was prevented by spinal cord transection at C1 with arterial pressure maintained by i.v. infusion of phenylephrine. After cord transection, CBF changed linearly with changes in arterial pco2 (r = 0.93; n = 23). FN stimulation (50-100 microA, 50 Hz, 1 s on/1 s off) produced sustained increases in CBF that developed slowly, reaching 50% of maximum within 24 +/- 1 s of stimulation (n = 17). After stimulation, CBF returned to baseline gradually within a time period (84-540 s) proportional to the duration of the stimulation (r = 0.93; n = 15). The CBF response was stimulus frequency and intensity dependent, was elicited only from restricted sites in FN, and was abolished by atropine (1 mg/kg, i.v.) or pentobarbital (30 mg/kg, i.v.). The slow temporal profile of the cerebrovasodilation is compatible with the hypothesis that in cerebral cortex local neurons mediate the vasodilation by interstitial release of vasoactive agents rather than by a direct action of neural processes on blood vessels. LDF is an effective technique for monitoring phasic change in CBF and may be useful in studies of the intrinsic neurogenic control of the cerebral circulation.