Preparation and absolute configuration of (1R,4R)-(+)-3-oxo-, (1S,4S)-(-)-3-oxo- and (1R,3S,4R)-(+)-3-acetyloxy-5-oxo-1 ,8-cineole.
Enantiomerically pure (1S,4S)-(-)-3-oxo-1,8-cineole (-)-2 and (1R,4R)-(+)-3-oxo-1,8-cineole (+)-2 were prepared for the first time and their absolute configurations assigned by vibrational circular dichroism (VCD) measurements. Thus, treatment of cineole 1 with chromyl acetate gave rac-2 which after sodium borohydride reduction and acetylation provided racemic 3-endo-acetyloxy-1,8-cineole, rac-4. Enantioselective hydrolysis using porcine liver esterase (PLE) gave a mixture of 3-endo-hydroxy-1,8-cineole (-)-3 and 3-endo-acetyloxy-1,8-cineole (+)-4. After chromatographic separation, (-)-3 was oxidized to (+)-2, while (+)-4 was hydrolysed to (+)-3 and then oxidized to (-)-2. The absolute configuration of either ketone 2 was established by VCD spectroscopy in combination with density functional theory (DFT) calculations at the B3LYP/DGDZVP level of theory, from where it followed that the (+)-2 enantiomer corresponds to (1R,4R)-1,3,3-trimethyl-5-oxo-2-oxabicyclo[2.2.2]octane and the (-)-2 enantiomer to the (1S,4S) molecule which is also in agreement with the absolute configuration deduced by the Mosher method for the starting chiral alcohols. Some literature inconsistencies are clarified. In addition, the enantiomerically pure monoester (1S,3S,4R,5R)-(-)-3-acetyloxy-5-hydroxy-1,8-cineole 6 and the ketoester (1R,3S,4R)-(+)-3-acetyloxy-5-oxo-1,8-cineole 7 were prepared from meso-diacetate 5 by enantioselective asymmetrization also using PLE.


A study of age at onset and affective temperaments in a Norwegian sample of patients with mood disorders.
Early age at onset of bipolar disorder is clinically important since it predicts a more severe course of illness and a poorer prognosis. Recently the age at onset (AAO) of bipolar disorder (BPD) has been reported different in samples from the USA and Europe (Bipolar Collaborative Network). We have examined the AAO of first major affective episode in major depressive disorder (MDD) and bipolar disorder in a sample of patients consecutively admitted to an affective ward in Norway. We also examined the relation between AAO and affective temperaments. Semi-structured interview of 119 patients consecutively admitted to an affective ward. Patients were diagnosed according to DSM-IV criteria for major affective disorders and affective temperaments were assessed using Akiskal Mallya criteria. Childhood-onset of BPD (before age 13) was found in 13.5% of patients, and early onset BPD (before age 20) in 61.6%. The AAO of first major affective disorder was significantly higher in MDD (n=67) compared to BPD (n=52); (28 Years Median (Min 10-Max 57) vs. 18 Years Median (Min 8-Max 41), p<0.001, Mann-Whitney U). Comparing patients with and without affective temperaments, showed that patients with such temperaments had an earlier AAO in both MDD and BPD (MDD: 20 Years Median (Min 10-Max 56), vs. 30 Years Median (Min 12-Max 57) p=0.006, Mann-Whitney U); BPD: 16 Years Median (Min 8-Max 30) vs. 20 Years Median (Min 12-Max 41, p=0.011, Mann-Whitney U). Our Norwegian data are more in line with the US than the European data regarding age at onset of bipolar disorders. The presence of an affective temperament presents an additional affirmation of the early occurrence of mood symptoms in early onset forms of both MDD and BPD.


Determination of the refractive index and thickness of transparent pellicles by use of the polarization-independent absentee-layer condition.
The refractive index and the thickness of a transparent pellicle are determined when the pellicle is placed between two vertical crossed polarizers and rotated in the horizontal plane. The transmission axes of the polarizers are neither parallel nor perpendicular to the plane of incidence. The light transmitted through the crossed polarizers reaches a minimum when the pellicle satisfies the absentee-layer condition. The refractive index and the film thickness are obtained from the pellicle orientation angles under such a condition.


In vitro cell culture pO2 is significantly different from incubator pO2.
Continuous noninvasive monitoring of peri-cellular liquid phase pO2 in adherent cultures is described. For neurons and astrocytes, this approach demonstrates that there is a significant difference between predicted and observed liquid phase pO2. Particularly at low gas phase pO2s, cell metabolism shifts liquid phase pO2 significantly lower than would be predicted from the O2 gas/air equilibrium coefficient, indicating that the cellular oxygen uptake rate exceeds the oxygen diffusion rate. The results demonstrate the need for direct pO2 measurements at the peri-cellular level, and question the widely adopted current practice of relying on setting the incubator gas phase level as means of controlling pericellular oxygen tension, particularly in static culture systems that are oxygen mass transfer limited.


Effects of potassium adaptation on blood pressure and pressor responses in normotensive and renal hypertensive Wistar rats.
Potassium adaptation reduces blood pressure (BP) in hypertensive humans and animals but its effects on normotensive BP and the nature of pressor responses to vasoactive drugs are not known. We measured directly, the mean arterial pressure (MAP) of normotensive control, normotensive potassium-adapted (given 0.75% potassium chloride solution for 5 weeks), renal hypertensive (RHP), and renal hypertensive Wistar rats later adapted to potassium. The maximum percentage change, the ED25, and recovery times after bolus injections of noradrenaline (NA), angiotensin II (Ang. II), sodium nitroprusside (SNP), and acetylcholine (ACh) were compared. The MAP of normotensive potassium-adapted rats was significantly lower than that of the normotensive controls (95.6+/-5.0 vs. 110.8+/-2.8 mmHg, p<0.05). The potassium-adapted hypertensive rats (RHP-A) also had significantly lower MAP values than the non-adapted hypertensive ones (116.0+/-4.4 vs. 138.2+/-4.1 mmHg, p<0.01). Potassium adaptation significantly blunted responses to NA and augmented responses to SNP but while the duration of action of Ang. II was significantly shortened, that of SNP was significantly increased. We conclude that potassium adaptation reduces BP in the normotensive and hypertensive rats and may influence both the degree and duration of action of vasoactive drugs given as bolus injections.


Single breath N2-test and exhaled nitric oxide in men.
The N(2) slope is an index of inhomogeneous distribution of ventilation and has been suggested to be suited for early testing of chronic obstructive pulmonary disease (COPD) in smokers. The aim of the present study was to examine the association between the fraction of exhaled nitric oxide (FENO) and the N(2) slope in a random population of smoking and non-smoking men. Altogether 57 subjects were included in the study, 24 never-smokers, seven ex-smokers and 26 current smokers. Subjects were examined twice, in 1995 when they regarded themselves as healthy, and in a follow-up in 2001. Spirometry, N(2) slope and high-resolution computed tomography (HRCT) were performed in 1995 while the follow-up examination included also measurement of FENO. The FENO value was significantly lower and the N(2) slope higher in current smokers. In smokers but not in never- or ex-smokers FENO was correlated to the difference in N(2) slope between 1995 and 2001 (r(s)=0.49, P=0.01). We analysed the data by multiple linear regression adjusted for smoking, mild respiratory symptoms and inhaled steroids. There were significant associations between FENO and the N(2) slope both in 1995 and in 2001. The strongest association was found to exist with the change in N(2) slope during these years. Sixteen of the subjects could be classified as having COPD, six with mild and ten with moderate COPD. There was a trend for an increase in N(2) slope with increased severity of COPD; among subjects with no COPD the N(2) slope in 2001 was 2.3% N(2)/L, and those with mild and moderate COPD had 2.5% N(2)/L and 3.9% N(2)/L, respectively (P=0.0004). No such trend was seen for FENO (17.8, 15.5 and 20.3 parts per billion (ppb), respectively, P=0.8). The results show that FENO is associated with the N(2) slope, indicating that FENO reflects inflammatory changes in the peripheral airways of both non-smoking and smoking subjects.


Cardiopulmonary bypass technique for treatment of renal cell carcinoma extending into the vena cava.
Cardiopulmonary bypass (CPB) is a unique technique used widely for renal cell carcinoma patients showing inferior vena cava invasion. We used a modification of CPB technique for 2 patients. These patients were managed with CPB without cross clamping and so without cold potassium cardioplegia. The primary tumor was a renal cell carcinoma propagating into the inferior vena cava and going into the right atrium as a thrombus in both patients. As a surgical procedure the urology team did nephrectomy and then with our clinical techniques we used CPB to cool the patients to 20 degrees C and decreased the flow to 500 ml/min/m2. The heart went into spontaneous ventricular fibrillation without using cross clamping and cardioplegia. Then we did atrial and inferior vena caval thrombectomy in a bloodless and visible operation field within a safe time interval. The aim of using this technique is to prevent myocardial injury and to protect the brain from hypoxia by using this low flow technique. This method can be used safely for the management of renal cell carcinomas and for some retroperitoneal malignancies associated with vena caval and atrial involvement.


The enhancement of photoresponse of an ordered inorganic-organic hybrid architecture by increasing interfacial contacts.
A modified ZnO quantum dot/polythiophene (ZnO/PTh) inorganic-organic hybrid architecture was fabricated by using ordered mesoporous silica (SBA-15) as the retaining template. First, a two-step strategy was developed to synthesize an ordered organic conducting polymer composite (PTh/SBA-15). Then, ZnO quantum dots were in situ formed on the pore walls of the ordered PTh/SBA-15 composite. Photoresponse of the inorganic-organic hybrid was studied with respect to its incident photon to collected electron conversion efficiency (IPCE) and morphology. The presence of SBA-15 proved to be critical for controlling the interfacial morphology and hence enlarging the interfacial area of the inorganic-organic heterojunction. The proposed approach may act as a key method to open up potential applications in photovoltaic devices.


Association of interleukin-10 polymorphisms with risk factors of Alzheimer's disease and other dementias (SADEM study).
Some studies have reported a genetic association between single nucleotide polymorphisms (SNPs) in the promoter region of Interleukin (IL) 10 and Alzheimer's disease (AD), with conflicting results. To further investigate the proposed association and to clarify the role of cytokines as a potential cause for AD susceptibility, we analyzed genotypes, allele distributions and haplotypes of IL-10 promoter polymorphisms -1082 (rs1800896) and -819 (rs1800871) in a Mexican population: 986 normal controls and 221 cases divided as follows: 122 with Alzheimer disease (AD), 67 with (VaD) and 32 with mixed dementia (AD/VaD). Patients with dementia showed increased frequency of "ATA, CTG, and CTA" haplotypes when compared to controls. We identified two risk haplotypes: ATA (OR=3.56, 95%CI=2.84-4.45, p<0.0001), and CTA (OR=1.90, 95%CI=1.38-2.62, p<0.0001), and four protection haplotypes: ATG (OR=0.60, 95%CI=0.45-0.82, p=0.0012), CTG (OR=0.38, 95%CI=0.23-0.62, p<0.0001), ACG (OR=0.01, 95%CI=0.002-1.13, p<0.0001), and CCG (OR=0.02, 95%CI=0.004-0.203, p<0.0001). In summary, this is the first study in Mexican population that considers the analysis of IL-10 in patients with AD, VaD and AD/VaD. Our results showed the relevance of the role that IL-10 plays in the pathological mechanisms that result in the development of dementia. In addition, in our study, it was possible to distinguish two protective and two risk haplotypes for the development of dementia.


[Dental students' opinions on education in fabricating single-tooth fixed prostheses and multi-teeth fixed prostheses].
Education in fabricating single-tooth fixed prostheses and multi-teeth fixed prostheses is an important component of the dental curriculum. To explore the opinion of dental students on this component of the curriculum, a web-based survey was carried out among the students of the 3 dental schools in the Netherlands. The 389 completed questionnaires revealed that the education in fabricating single-tooth fixed prostheses and multi-teeth fixed prostheses started in different years of study at the 3 dental schools. Another striking difference is that the methods of preparation and the choice of materials for single-tooth fixed prostheses and multi-teeth fixed prostheses in the pre-clinical practicum in the programmes was not the same. The students of the 3 dental schools seemed to have a reasonably positive opinion of their preclinical training preparation for treating patients. The opinions of the Nijmegen Dental School students were significantly more positive than the opinions of the students at the 2 other dental schools.


Identification and Biosynthesis of 1-Mercaptoethanesulfonic Acid (1-MES), an Analogue of Coenzyme M, Found Widely in the Methanogenic Archaea.
Here I report on the identification of 1-mercaptoethanesulfonic acid (1-MES), an analogue of 2-mercaptoethanesulfonic acid (coenzyme M, HSCoM). 1-MES and HSCoM were both present in the growth media of eight different methanogens at concentrations ranging from ∼1 to 100 μM. In an effort to determine a chemical origin of 1-MES, several plausible chemical routes were examined each assuming that HSCoM was the precursor. In all examined routes, no 1-MES was formed. However, 1-MES was formed when a solution of vinylsulfonic acid and sulfide were exposed to ultraviolet light. On the basis of these results, I conclude 1-MES is formed enzymatically. This was confirmed by growing a culture of Methanococcus maripaludis S2 in the presence of [1,1',2,2'-2H4]HSCoM and measuring the incorporation of deuterium into 1-MES. 1-MES incorporated three of the four deuteriums from the fed HSCoM. This result is consistent with the abstraction of a C-2 deuterium of the HSCoM, likely by a 5'-dAdoCH2• radical, followed by a radical rearrangement in which the sulfonic acid moves to position C-1, followed by abstraction of a H• likely from 5'-dAdoCH2D. At present, the reason for the production of 1-MES is not clear. This is the first report of the occurrence of 1-MES in Nature.


Specific heat of multifractal energy spectra.
Motivated by the self-similar character of energy spectra demonstrated for quasicrystals, we investigate the case of multifractal energy spectra, and compute the specific heat associated with simple archetypal forms of multifractal sets as generated by iterated maps. We considered the logistic map and the circle map at their threshold to chaos. Both examples show nontrivial structures associated with the scaling properties of their respective chaotic attractors. The specific heat displays generically log-periodic oscillations around a value that characterizes a single exponent, the "fractal dimension," of the distribution of energy levels close to the minimum value set to 0. It is shown that when the fractal dimension and the frequency of log oscillations of the density of states are large, the amplitude of the resulting log oscillation in the specific heat becomes much smaller than the log-periodic oscillation measured on the density of states.


Quantitative polymerase chain reaction analysis reveals marked overexpression of interleukin-1 beta, interleukin-1 and interferon-gamma mRNA in the lymph nodes of lupus-prone mice.
The nature of the stimuli driving autoantibody production in systemic lupus erythematosus (SLE) is unclear, but cytokines are believed to play an important role. Since cytokines primarily appear to act locally at the tissue level, we analysed mRNA expression of several cytokines (IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, IFN gamma, TNF alpha, TNF beta and TGF beta 1) in the lymph nodes of lupus-prone mice, in models of early onset disease. We constructed a multispecific competitor fragment that allowed quantification of these cytokine transcripts by competitive PCR assay. The results reveal considerable overexpression of IL-1 beta, IL-10 and IFN gamma transcripts in SLE-prone MRL-lpr/lpr (MRL/l) and BXSB male (BXSBm) mice, but with some strain differences. IFN gamma was most markedly augmented in MRL/l mice (in some cases over 100-fold greater than control mice), IL-1 beta was most severely overexpressed in BXSBm mice while IL-10 was equally increased in both strains. In addition, TGF beta 1 expression was moderately elevated in the lymph nodes of BXSBm (but not MRL/l) mice. We found no abnormality in the expression of the other cytokines. Cytokine transcript levels were only slightly altered at 4 weeks of age, but were elevated from 10 to 22 weeks of age. The latter phase corresponds to a period where lupus-like disease escalates, resulting in frequent mortality. Interestingly, our results do not reveal a clear Th1 or Th2 cytokine expression pattern in these lupus-prone mice. IL-1 beta, IFN gamma and IL-10 are pleiotropic cytokines with pro-inflammatory and B-cell stimulatory effects. These results point to certain cytokines as potential targets for immunotherapy in lupus.


[The changes in the oxygen saturations in the superior vena cava and the pulmonary artery are not the same during cardiac surgery].
To evaluate the changes over time (trend) in sign and magnitude for SSVO2 and SVO2 during and after cardiac surgery. A prospective and observational study was conducted on 34 cardiac surgery patients. Venous blood samples were taken simultaneously from the introductor (SVCO2) and distal (SVO2) port of the pulmonary artery catheter at predefined intervals. Systemic and pulmonary hemodynamic variables were measured at the same time. The trend was calculated as the difference between 2 consecutive measurements (tSO2). Data were processed with ANOVA for multiple comparisons, Pearson correlation coefficient and Bland-Altman analysis. There was a significant correlation between SVCO2 and tSVO2 (R(2)=0.55), the mean of the differences was 0.36±7.75%, and the limits of agreement ranged from -15.1 to 15.9%. The sign of the trend was similar in 85.1% of the paired data. However, the magnitude of the changes in tSVCO2 and tSVO2 were not always equivalent. Between 0 and 5% of the change in the tSVCO2 was coincident with only 44.7% of the tSVO2. A wide variation was found between both trends when the signs and magnitudes of the changes were taken into account. When considering the sign and magnitude, the change over time of central venous O2 saturations were not interchangeable in cardiac surgery patients. Clinical decisions based exclusively on tSVCO2 monitoring should be taken with caution.


The psychopathology of echophenomena.
Repetition of other person's activities is, to some extent, part of normal human behaviour. However, in some circumstances it becomes pathological and symptomatic of an underlying disease process. The pathogenesis of this type of behaviour is very diverse. The circumstances in which echoing can be expected to occur are reviewed.


Effects of Vepacide (Azadirachta indica) on aspartate and alanine aminotransferase profiles in a subchronic study with rats.
The aim of this study was to ascertain the long-term effects of Vepacide, a neem-based pesticide on biochemical profiles. Albino Wistar rats were treated orally with 80 (low), 160 (medium) and 320 mg/kg (high) doses of Vepacide in coconut oil for 90 days. Control rats received the same volume of the vehicle. Vepacide caused increase of aspartate and alanine aminotransferase in serum, kidney and lung, and these enzymes decreased in liver in both male and female rats when measured after 45 and 90 days of treatment. The two-way analysis of variance (ANOVA) showed that the alterations in these enzymes were dose- and time-dependent. Sexual dimorphism was observed when male rats were compared with female rats (Student t-test at P< 0.05). Positive correlation was observed with regard to these enzymes between serum, kidney and lung, whereas in the case of serum and liver, a negative correlation was recorded. These enzyme profiles elucidate that they increased in serum with simultaneous decrease in liver, indicating necrosis of liver, whereas in other tissues, the level of enzymes increased, showing an adaptive mechanism due to the chemical stress. The affected enzymes were recovered to normal conditions after 28 days of post-treatment (withdrawal study). Due to the Vepacide treatment, lung was more affected followed by liver and kidney. This study has indicated that these enzymes could be useful as biomarkers for the insult of any toxicant. Besides, they can also help in predictive toxicology.


Contemporary protease inhibitors and cardiovascular risk.
To review the evidence linking use of HIV protease inhibitors with excess risk of cardiovascular disease (CVD) in HIV+ populations. For the two contemporary most frequently used protease inhibitors, darunavir and atazanavir [both pharmacologically boosted with ritonavir (/r)], darunavir/r has been shown to be associated with increased CVD risk. The effect is cumulative with longer exposure increasing risk and an effect size comparable to what has been observed for previously developed protease inhibitors. Biological mechanisms may be overlapping and include perturbed lipid metabolism and accumulation of cholesterol derivatives within macrophages. Conversely, atazanavir/r has not been shown to be associated with CVD, possibly because of its ability to increase cardioprotective bilirubin levels. Evidence linking protease inhibitors to CVD is based on observational studies only, whereas plausible biological explanations are well established and derived from randomized trials and controlled experiments. Given the possible association with clinical disease, a conservative approach to apply the data in daily practise is proposed which is focused on individualization of care based on underlying risk of CVD.


Senior nurses to be groomed for CNO post.
The Department of Health has picked six senior nurses for a 'high potential' leadership programme that aims to groom the next chief nursing officer (CNO) for England.


Treatment of scapho-lunate instability.
Scapho-lunate dissociation is one of the most frequent reasons of carpal instability. Diagnosis is mostly based on a careful physical examination where stability of the SL joint is specifically checked through different clinical maneuvers, followed by static and dynamic radiographic examination of the wrist in a variety of positions and loading conditions aiming at demonstrating the presence of a gap between the two dissociated bones. When in doubt, arthroscopy is used. In the treatment five factors are to be considered: integrity of the dorsal SL ligament, its healing potential, status of other scaphoid stabilizers, reducibility of malaligment and cartilage status. The staging classification of SLD is also presented. On this basis the possible treatment techniques both recent and past are described, including various methods of soft tissue repair and reconstruction procedures and bony stabilization.


A method to quantify the uptake rate of 2-[18F]fluoro-2-deoxy-D-glucose in tissues.
A two-compartment model is used to derive the tissue time-activity curve of 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG). At the later phase of the 18F-FDG activity in the tissue, the model leads to a method which allows the assessment of the 18F-FDG uptake rate constant. The proposed method is both compared with the standard uptake value (SUV) method and with Patlak's graphical analysis.


Double expressions of connexin 43 and 32 in human periodontal ligament fibroblasts.
The expressions of connexin 43 and 32 in cultured and intact human periodontal ligament fibroblasts (PDLFs) were examined using immunohistochemical methods, and western blot analysis was conducted with anti-connexin 43 and 32 in cultured PDLFs. The PDLFs both in cultured cells and tissue sections reacted with anti-connexin 43 and 32, and western blot analysis showed bands of approximately 43 kD and 27 kD reacted with anti-connexin 43 and 32 respectively, suggesting the existence of gap junctions in human PDLFs. In cultured PDLFs there were no reaction products of connexin 43 when the cells were not in contact with adjacent cells, but reaction products were increasingly observed with increases in cell-cell contacts. Different from connexin 43, the reaction products of connexin 32 were found in the cytoplasm, regardless of whether the cells were or were not in contact with adjacent cells. Further, the reaction activity of connexin 32 varied among PDLFs; some were strong, some moderate, and some weak. The expressions of connexin 43 and 32 in human PDLFs are suggested to be related to the regulation of two different functions of the PDLFs.


Biatrial neuroablation attenuates atrial remodeling and vulnerability to atrial fibrillation in canine chronic rapid atrial pacing.
We investigated the proposition that an intact cardiac nervous system may contribute to electrophysiological remodeling and increased vulnerability to atrial fibrillation (AF) following chronic rapid atrial pacing (RAP). Baseline study was conducted prior to ablating right and left ganglionated plexuses (RAGP, LAGP) in 11 anesthetized canines (Neuroablation group) and in 11 canines without neuroablation (Intact GP). After being subjected to RAP (400 beats/min) for 6 weeks, animals were reanesthetized for terminal study. The ERP shortening typical of chronic RAP was significantly more pronounced in the Intact GP (baseline: 112 ± 12 to terminal: 80 ± 11 ms) than in the Neuroablation group (113 ± 18 to 102 ± 21 ms, p < .001), and AF inducibility (extrastimulus protocol) showed significantly greater increment in the Intact GP (baseline: 23 ± 19% to terminal: 60 ± 17% of trials) than in the Neuroablation group (18 ± 15% to 27 ± 17%, p = 0.029). Negative chronotropic responses to right vagus nerve stimulation were markedly reduced immediately after the neuroablation procedure but had recovered at terminal study. Vagally-evoked repolarization changes (from 191 unipolar electrograms) occurred in a majority of Intact GP animals in the superior, middle and inferior RA free wall, and in the LA appendage. In the Neuroablation group, repolarization changes were restricted to the superior RA free wall but none occurred in the inferior RA and only infrequently in the LA appendage, yielding significantly smaller affected areas in Neuroablation than in Intact GP animals. Persistent functional denervation in LA and RA regions other than RA pacemaker areas may contribute to prevent the development of a tachycardia-dependent AF substrate.


The fine structure of the placenta and chorionic vesicles of the bush baby, Galago crassicaudata.
The fine structure of the placental villi and the chorionic vesicles of the bush baby, Galago crassicaudata, were examined. The placenta was of the diffuse epitheliochorial type. The trophoblastic epithelium of the placental villi consisted of cells joined by tight junctions and desmosomes. The apical borders of these cells interdigitated with corresponding processes on the uterine epithelial cells. The fetal capillaries indented the trophoblastic cells later in gestation, and the trophoblast over the capillaries was correspondingly thinner with advancing gestation. The trophoblast of the villi was characterized by numerous lipid droplets and moderate amounts of granular endoplasmic reticulum (ER). The apical cytoplasm often had many mitochondria as well as small electron-dense vesicles and tubules. Uterine epithelial cells contained agranular ER and a well-developed Golgi apparatus. The apical cytoplasm contained numerous granules often with an electron-dense content. The uterine epithelial cells appeared to be secretory, contributing to the electron-dense material found between the fetal and maternal cells. Fetal capillaries were of the continuous type, whereas occasional fenestrae were observed in maternal capillaries. Recesses in the trophoblastic epithelium occurred at the tips, sides, and bases of the villi. These areas had taller trophoblastic cells that appeared to be more active in phagocytosing uterine content. The chorionic vesicles are invaginations of the chorion opposite the mouths of uterine glands. A layer of columnar trophoblastic cells covered the villi of the vesicles. These cells had abundant vacuoles and coated pits and vesicles. The mesodermal component of the chorionic vesicles included a capillary network and a layer of smooth muscle cells in the wall of the vesicle. It was concluded that the trophoblast of the placental villi is engaged in both hemotrophic and histotrophic nourishment of the embryo. The specialized chorionic vesicles are particularly important in providing histotrophic nutrients to the embryo, especially the secretions of the uterine glands. The similarity of the chorionic vesicles to the areolae of other species suggests they may be the pathway by which iron is transported from mother to fetus, perhaps by receptor-mediated endocytosis of macromolecules derived from uterine gland secretions.


The value of cerebrospinal fluid antiviral antibody in the diagnosis of neurologic disease produced by varicella zoster virus.
We studied four patients with subacute to chronic varicella zoster virus (VZV) infection of the central nervous system (CNS). VZV infection was verified by detecting antibody to VZV in the cerebrospinal fluid (CSF). VZV caused myelitis in two patients and encephalitis in two patients. In one of the patients with VZV encephalitis, in addition to VZV IgM antibody, VZV DNA was found in the CSF. Among the four patients with VZV infection of the CNS, CSF antibody to VZV was the crucial diagnostic laboratory test which corroborated the clinical features, and indicated that VZV caused neurologic disease. In addition to looking for amplifiable VZV DNA in the CSF of patients with neurologic disease whose clinical and radiologic features suggest VZV infection, we also recommend a search for CSF antibody to VZV, particularly in patients with intervals of weeks to months between zoster and the onset of neurologic disease, or in those patients without rash in whom the tempo of illness is unknown.


The minimum infusion rate of alfaxalone during its co-administration with lidocaine at three different doses by constant rate infusion in goats.
To determine the minimum infusion rate (MIR) of alfaxalone required to prevent purposeful movement in response to standardized stimulation while co-administered with lidocaine at three different doses by constant infusion rate infusion (CRI) in goats. Prospective, blinded, randomized crossover, experimental. A total of eight healthy goats: four does and four wethers. Anaesthetic induction was with lidocaine at 1 mg kg-1 [low dose of lidocaine (L-Lid)], 2 mg kg-1 [moderate dose (M-Lid)] or 4 mg kg-1 [high dose (H-Lid)] and alfaxalone at 2 mg kg-1. Anaesthetic maintenance was with alfaxalone initially at 9.6 mg kg-1 hour-1 combined with one of three lidocaine treatments: 3 mg kg-1 hour-1 (L-Lid), 6 mg kg-1 hour-1 (M-Lid) or 12 mg kg-1 hour-1 (H-Lid). The MIR of alfaxalone was determined by testing for responses to a stimulation in the form of clamping on a digit with a Vulsellum forceps every 30 minutes during lidocaine CRI. Basic cardiopulmonary parameters were measured. The alfaxalone MIRs were 8.64 (6.72-10.56), 6.72 (6.72-8.64) and 6.72 (6.72-6.72) mg kg-1 hour-1 during L-Lid, M-Lid and H-Lid, respectively, without any significant differences among treatments. Compared to the initial rate of 9.6 mg kg-1 hour-1, these reductions in MIR are equivalent to 10, 30 and 30%, respectively. Significant increases in heart rate (HR) and arterial carbon dioxide partial pressure (PaCO2) and decreases in arterial haemoglobin saturation (SaO2), arterial oxygen partial pressure (PaO2) and respiratory frequency (fR) immediately after induction were observed during all lidocaine treatments. Lidocaine reduces the alfaxalone MIR by up to 30% with a tendency towards a plateauing in this effect at high CRIs. Immediate oxygen supplementation might be required to prevent hypoxaemia.


Stimulation of skin menthol receptors stimulates detrusor activity in conscious rats.
The Transient Receptor Potential Melastatin (TRPM)8 channel, activated by menthol and cool temperatures, has been proposed to be a cool receptor involved in mediation of the bladder response in the diagnostic ice water test. We examined the expression of TRPM8 in the rat skin and investigated if stimulation of skin menthol receptors by menthol can induce detrusor activity in conscious rats. Immunofluorescence staining for TRPM8 was performed in samples of the skin from the leg and back of the rats. Continuous cystometry was performed in conscious animals. Voiding interval (VI), micturition volume (MV), and bladder capacity (BC) were evaluated before and after spraying menthol solution to the skin of the leg and back of the rats. Immunohistochemical analysis revealed that TRPM8 was expressed in the investigated tissues. The expression of TRPM8-positive receptors was not significantly different between the leg and back skin. Saline caused no significant change in cystometric parameters. After spraying menthol solution (50% and 99% to the skin of the leg, and 99% to the back skin), VI, MV, and BC decreased significantly. TRPM8 is expressed in rat skin. Spraying menthol solution onto the skin-induced detrusor activity, an effect we suggest is mediated by stimulation of TRPM8 receptors.


Experiences using the Fixin locking plate system for the stabilization of appendicular fractures in dogs: a clinical and radiographic retrospective assessment.
To retrospectively evaluate the stabilization of appendicular fractures in dogs using the Fixin locking plate system. Medical records and radiographs of dogs with fractures stabilized with the Fixin system in the period from May 2005 to September 2010 were reviewed. For each patient, data pertaining to signalment, the nature of the fracture, implants used, and evidence of fracture healing were recorded. The outcome and complications were determined from clinical and radiographic follow-up examinations. Limb function was evaluated between 40 days and 90 days postoperatively. Owners of pets with complications were contacted by phone for long-term follow-up. Eighty-two fractures in seventy-five dogs met the inclusion criteria for the study. Radiographic re-examinations were carried out between eight days to two years (median 60 days) following surgery. Seventy-three out of 82 fractures (89%) reached union without complications. Major complications were seen in six dogs (7%). Limb function was graded as 'normal' in 73/75 (97%) dogs and 'mild lameness' in 2/75 (3%) cases. Despite a modest complication rate, the Fixin locking bone plating system appears to be an acceptable choice of implant for the stabilization of appendicular fractures in dogs.


Enzymatic asymmetric synthesis by decarboxylases.
Decarboxylation reactions using microbial cells or enzymes are increasingly being used for the synthesis of enantiomerically pure compounds because of their high degree of regio- and stereo-specificity. Pyruvate decarboxylase, benzoylformate decarboxylase and phenylpyruvate decarboxylase enzymes are capable of acyloin-type condensation reactions leading to formation of chiral alpha-hydroxy ketones, which are versatile building blocks in the pharmaceutical and chemical industries. Availability of three-dimensional structures of some decarboxylases in recent years has facilitated understanding of reaction mechanisms and the creation of mutants with enhanced activity and stability.


Alternative end-joining pathway(s): bricolage at DNA breaks.
To cope with DNA double strand break (DSB) genotoxicity, cells have evolved two main repair pathways: homologous recombination which uses homologous DNA sequences as repair templates, and non-homologous Ku-dependent end-joining involving direct sealing of DSB ends by DNA ligase IV (Lig4). During the last two decades a third player most commonly named alternative end-joining (A-EJ) has emerged, which is defined as any Ku- or Lig4-independent end-joining process. A-EJ increasingly appears as a highly error-prone bricolage on DSBs and despite expanding exploration, it still escapes full characterization. In the present review, we discuss the mechanism and regulation of A-EJ as well as its biological relevance under physiological and pathological situations, with a particular emphasis on chromosomal instability and cancer. Whether or not it is a genuine DSB repair pathway, A-EJ is emerging as an important cellular process and understanding A-EJ will certainly be a major challenge for the coming years.


Epidemiology of contact lens-related inflammation and microbial keratitis: a 20-year perspective.
Contact lens-related keratitis has been the subject of a number of rigorous epidemiologic studies over the past 20 years. This article summarizes data from such studies that address incidence and risk factors for this condition. Estimates of risk and associated predisposing factors have been quite consistent over time, geography and study design.


Chemical composition of the essential oil of leaves and roots of Ottoa oenanthoides (Apiaceae) from Mérida, venezuela.
Essential oils extracted by hydrodistillation from leaves and roots of Ottoa oenanthoides Kunth (Apiaceae) were analyzed by GC/MS. The oils, obtained in yields of 0.10% and 0.66%, respectively, each contained four compounds, which were identified from their mass spectra and retention indices (RI). The major compound identified was 2-methoxy-8-methyl-1,4-naphthalindione (59.9% leaves, and 62.8%, roots), followed by 7-methoxy-1-naphthol (18.3% leaves and 17.3% roots), 2-naphthalenol (18.6% leaves and 15.0% roots), and 3-methoxy-2-naphthalenol (3.1% leaves and 2.1% roots). To the best of our knowledge, this is the first time that naphthalene derivatives have been reported for any species of the Apiaceae family.


A non-heme iron protein with heme tendencies: an investigation of the substrate specificity of thymine hydroxylase.
Thymine hydroxylase from Rhodotorula glutinis catalyzes the oxidation of thymine to its alcohol, aldehyde, and carboxylic acid in three successive reactions. Each step involves stoichiometric consumption of O2 and alpha-ketoglutarate and formation of CO2 and succinate. Given the promiscuity of this enzyme, it was hoped that it would serve as a prototype for understanding the mechanism of this class of enzymes, the non-heme Fe2+ dioxygenases. Kinetic parameters for thymine, O2, Fe2+, and alpha-ketoglutarate have been determined, and isotope effect analysis of (trideuteriomethyl)thymine with enzyme reveals D(V) = 2.08 and D(V/K) = 1.11 at saturating O2. The kinetic parameters for (hydroxymethyl)uracil oxidation have been determined, and incubation of (5'-R)- and (5'-S)-[5'-2H]-5-(hydroxymethyl)uracil with enzyme reveals stereospecific removal of the pro-S hydrogen. No apparent isotope effect is observed in this reaction. The substrate specificity of this enzyme has been examined in detail. The enzyme can catalyze epoxidation, oxidation of a thioether to a sulfoxide and a sulfone, hydroxylation of an unactivated carbon-hydrogen bond, and oxidation of a methylamine to formaldehyde, as revealed through studies with 5-vinyluracil, 5-(methylthio)uracil, 5,6-dihydrothymine, and 1-methylthymine, respectively. In each case, the products were identified by gas chromatography-mass spectrometry, and 18O2-labeling studies revealed that one atom from O2 is incorporated into each product. The enzyme has also been shown to catalyze an uncoupling of hydroxylation and decarboxylation in the presence of a substrate analog incapable of undergoing hydroxylation or a substrate that is difficult to oxidize.(ABSTRACT TRUNCATED AT 250 WORDS)


Gastric carcinoma exhibits distinct types of cell differentiation: an immunohistochemical study of trefoil peptides (TFF1 and TFF2) and mucins (MUC1, MUC2, MUC5AC, and MUC6).
The expression of trefoil peptides (TFF1 and TFF2) and mucins (MUC1, MUC2, MUC5AC, and MUC6) has previously been described in gastric polyps. In the present study, the expression profile of these trefoil peptides and mucins was characterized in 96 gastric carcinomas, in an attempt to further the understanding of the histogenesis and cell differentiation of gastric carcinoma. Taking together the co-expression of trefoil peptides and mucins, three phenotypes were defined: complete gastric, incomplete gastric, and non-gastric phenotype. Gastric differentiation (complete and incomplete) was observed in 30 out of 33 (90.9%) diffuse carcinomas and in 38 out of 53 (71.7%) intestinal carcinomas. Non-gastric differentiation was observed in only three (9.1%) diffuse carcinomas and in 15 (28.3%) intestinal carcinomas. The phenotypes observed in intestinal carcinomas were similar to those previously observed in adenomatous polyps, whereas most diffuse carcinomas mimicked the phenotype of hyperplastic polyps. The percentage of cases displaying a non-gastric phenotype was higher, though not significantly, in tumours that had invaded the gastric wall than in T1 tumours, regardless of histotype. It is concluded that gastric-type differentiation is retained in the majority of gastric carcinomas, being more prominent in diffuse than in intestinal carcinomas, and in early than in advanced carcinomas.


[The effect of supplying rats with vitamin K and administration of pelentane on Na,K- and spectrin-dependent ATPase in erythrocyte ghosts].
It was found that the activity of spectrin-dependent ATPase of erythrocyte ghosts isolated from rats with alimentary deficiency of vitamin K was significantly increased as compared with control animals, whereas in rats kept on a vicasol-rich diet this parameter was unchanged. In vitamin K-deficient rats the amount of proteins loosely bound to erythrocyte membranes was significantly reduced. At the same time, the activity of the integral enzyme (Na, K-ATPase) did not depend on the vitamin K provision despite the fact that in vitamin K-deficient animals kept on a vicasol-rich diet the enzyme affinity for ouabain was strongly decreased as compared with control. It was suggested that this effect might be due to the changes in the lipid and protein environment of the membrane-bound enzyme. Administration of the antivitamin K, pelentane, did not induce any conspicuous changes in the enzyme activities. It was concluded that antivitamin K does not induce any modification of the properties of erythrocyte-linked enzymes observed under conditions of vitamin K deficiency.


Involvement of occupational therapy departments in research: a provincial survey.
It is increasingly recognized that research is necessary to advance the practice of occupational therapy. The objective of this study was to determine whether occupational therapy departments in Quebec were commonly involved in research, and whether participation varied with the size of department and type of work setting. Secondly, enablers and barriers to participation in research were identified. A random sample of occupational therapy departments, stratified by size, were surveyed by mail. Participation rate was 74.8% (107/143). Participation in research was noted in half of the departments surveyed (51/107), most frequently as collaborator. Involvement in research was associated (p < .001) with a greater number of therapists in the department. Furthermore, occupational therapy departments in rehabilitation centres and in university hospitals were more likely to participate in research (79% and 90% respectively), whereas participation was lower for long-term care facilities (29%) and community health clinics (38%). When asked to rank factors that may facilitate participation in research, the highest rankings were given to: time allotted to research, research as a priority for administration, adequate financial support, and presence of an occupational therapy researcher and a research centre on site. This survey demonstrates that a number of factors can promote or prevent the realization of research activities within the clinical setting. Strategies are proposed to enhance the integration of research into clinical practice.


Preliminary X-ray data analysis of crystalline cowpea chlorotic mottle virus.
Crystals of cowpea chlorotic mottle virus (CCMV) that diffract X-rays to 3.1 A resolution were grown in a succinate-PEG solution buffered at pH 3.3. The crystals are in space group P2(1)2(1)2(1) with unit cell dimensions of a = 381.26 A, b = 381.26 A, and c = 408.59 A. Four particles occupy the unit cell, placing a single virion in the crystallographic asymmetric unit. Diffraction intensities measured from 196 films collected at the Cornell High Energy Synchrotron Source accounted for 55% of the theoretically possible data to 3.2 A. Unit cell dimensions and rotation function analyses of the X-ray data revealed that the particles were organized in a pseudo-tetragonal relationship with the pseudo-fourfold axis along the crystal c axis. Analysis of electron micrographs of two-dimensional crystals of CCMV revealed a remarkable similarity between these and planes of particles perpendicular to the crystallographic c axis in the three-dimensional crystal.


Differential response of Apis mellifera acetylcholinesterase towards pirimicarb.
The kinetic analysis of Apis mellifera acetylcholinesterase inhibition by the carbamate pirimicarb showed that native and detergent-solubilized membrane enzyme exhibited slightly different carbamylation kinetics. The acetylcholinesterase form sensitive to phosphatidylinositol-specific phospholipase C (PI-PLC) was carbamylated more rapidly (kapp = 36.4 X 10(-3) min-1) than the PI-PLC-resistant counterpart (kapp = 10.13 X 10(-3) min-1) which had a behavior close to that of the soluble tryptic enzyme (kapp = 11.89 X 10(-3) min-1). A difference in acetylcholinesterase sensitivity towards pirimicarb was also observed between foraging and emerging bees. These results show that the molecular structure, the mode of preparation and the source of acetylcholinesterase from the bee head should be taken into account in accurate toxicological studies.


Multiple-life-stage probabilistic risk assessment for the exposure of Chinese population to PBDEs and risk managements.
Studies assessing body burden of polybrominated diphenyl ethers (PBDEs) exposure have been conducted in the United States and Europe. However, the long-term assessment that is associated with multimedia exposure of PBDEs for the Chinese population is not available. The current study estimated the health risks using large PBDEs data to quantify the contributions of various media from different regions and distinguished the most vulnerable periods in life. We summarized media-specific (soil, dust, outdoor and indoor air, human milk and food) concentration of PBDEs in China from 2005 to 2016. Probabilistic risk assessment was adopted to estimate the health risks of infants, toddlers, children, teenagers and adults through ingestion, inhalation and dermal absorption. Monte Carlo simulation and sensitivity analysis were performed to quantify risk estimates uncertainties. E-waste areas had the highest PBDEs concentration, which was at least an order of magnitude higher than in other areas. BDE209 was the primary congener, accounting for 38-99% of the estimated daily intake. The dominant exposure pathway for infants was dietary intake through human milk, whereas dust ingestion was a higher contributing factor for toddlers, children, teenagers and adults. The 95th percentile of hazard index for infants and toddlers from e-waste areas of Guangdong and Zhejiang provinces exceeded one. Our estimates also suggested that infants may have the highest body burdens of PBDEs compared to other age groups. Sensitivity analyses indicated that PBDEs concentrations and ingestion rates contributed to major variances in the risk model. In this study, e-waste was found as a significant source of PBDEs, and PBDEs-containing e-waste are likely to be a threat to human health especially during early period of life. Risk strategies for better managing environmental PBDEs-exposure and human health are needed, due to the high intake of PBDEs and their persistence in the environment.


Identifying structural motifs in proteins.
In biological macromolecules, structural patterns (motifs) are often repeated across different molecules. Detection of these common motifs in a new molecule can provide useful clues to the functional properties of such a molecule. We formulate the problem of identifying a given structural motif (pattern) in a target protein (example) and discuss the notion of complete matches vis-a-vis partial matches. We describe the precise error criterion that has to be minimized and also discuss different metrics for evaluating the quality of partial matches. Secondly, we present a new polynomial time algorithm for the problem of matching a given motif in a target protein. We also use the sequence and (if available) secondary structure information to annotate the different points in motif and the target protein, thus reducing the search space size. Our algorithm guarantees the detection of a perfect match, if present. Even otherwise, the algorithm computes very good matches. Unlike other methods, the error minimized by our algorithm directly translates to root mean square deviation (RMSD), the most commonly accepted metric for structure matching in biological macromolecules. The algorithm does not involve any preprocessing and is suitable for the detection of both small and large motifs in the target protein. We also present experiments exploring the quality of matches found by the algorithm. We examine its performance in matching (both full and partial) active sites in proteins.


Management of hypertensive emergencies in children.
Pediatric hypertension is considered a relatively recent disease. Most hypertensive children do not require emergent care and can be managed in an outpatient setting. On the other hand, hypertensive emergencies and urgencies which can be encountered in the outpatient clinics, the inpatient wards or during an emergency visit need prompt recognition and treatment. Efficient management of these children is essential to avoid some of the life-threatening complications associated with hypertension and its treatment. This article will focus on the main pharmacologic agents used in the management of hypertensive emergencies in the pediatric population.


Multidetector CT of solitary pulmonary nodules.
With the increasing use of multidetector CT, small nodules are being detected more often. Although most incidentally discovered nodules are benign, usually the sequelae of pulmonary infection and malignancy, either primary or secondary, remains an important consideration in the differential diagnosis of solitary pulmonary nodules. This article reviews the role of imaging in the detection and characterization of solitary pulmonary nodules. Strategies for evaluating and managing solitary pulmonary nodules are also discussed.


Bioaccumulation and risk assessment of per- and polyfluoroalkyl substances in wild freshwater fish from rivers in the Pearl River Delta region, South China.
Per- and polyfluoroalkyl substances (PFASs) are used in various industries, which results in their ubiquitous occurrence in the environment. This study determined the concentrations of eighteen PFASs in muscle and liver of nine wild freshwater fish species collected from rivers in the Pearl River Delta (PRD) region, South China, and assessed their bioaccumulation and potential health risks to local people. The results showed that eight and twelve PFASs were detected in the fish muscle and liver samples, respectively. Perfluorooctane sulfonate (PFOS) was found to be the predominant PFAS both in muscle and liver with its highest concentrations of 79ng/g wet weight (ww) in muscle and 1500ng/g ww in liver, followed by Perfluoroundecanoic acid (PFUnDA) and Perfluorotridecanoic acid (PFTrDA) with trace concentrations. The mean PFOS concentrations in fish muscle and liver tissues of the nine collected species ranged from 0.40ng/g in mud carp to 25ng/g in snakehead, and from 5.6ng/g in mud carp to 1100ng/g in snakehead, respectively. Significant positive correlations were found among PFASs both in water and fish, indicating a similar pollution source for these PFASs. In tilapia samples, PFOS concentrations showed an increasing trend with increasing length and weight, but no significant difference between genders. Bioaccumulation factors (logBAF) in fish for the PFASs were in the range from 2.1 to 5.0. The calculated hazard ratios (HR) of PFOS for all fishes were in the range of 0.05-2.8, with four out of nine species (tilapia, chub, leather catfish and snakehead) having their HR values more than 1.0. The results suggest that frequent consumption of these four fish species may pose health risks to local population.


[A message on the occasion of the 50th anniversary of the World Health Organization].
The Director of the Panamerican Health Organization gives us a message about the 50th anniversary of WHO, emphasizing its origin and characteristics, the successes of its programmes and policies to the sound management of health programmes in its member countries, through the strategy of primary health care and health for all programmes.


Effects of daily precipitation and evapotranspiration patterns on flow and VOC transport to groundwater along a watershed flow path.
MTBE and other volatile organic compounds (VOCs) are widely observed in shallow groundwater in the United States, especially in urban areas. Previous studies suggest that the atmosphere and/or nonpoint surficial sources could be responsible for some of those VOCs, especially in areas where there is net recharge to groundwater. However, in semiarid locations where annual potential evapotranspiration can exceed annual precipitation, VOC detections in groundwater can be frequent. VOC transport to groundwater under net discharge conditions has not previously been examined. A numerical model is used here to demonstrate that daily precipitation and evapotranspiration (ET) patterns can have a significant effect on recharge to groundwater, watertable elevations, and VOC transport. Ten-year precipitation/ET scenarios from six sites in the United States are examined using both actual daily observed values and "average" pulsed precipitation. MTBE and tetrachloroethylene transport, including gas-phase diffusion, are considered. The effects of the precipitation/ET scenarios on net recharge and groundwater flow are significant and complicated, especially under low-precipitation conditions when pulsed precipitation can significantly underestimate transport to groundwater. In addition to precipitation and evapotranspiration effects, location of VOC entry into the subsurface within the watershed is important for transport in groundwater. This is caused by groundwater hydraulics at the watershed scale as well as variations in ET within the watershed. The model results indicate that it is important to consider both daily precipitation/ET patterns and location within the watershed in order to interpret VOC occurrence in groundwater, especially in low-precipitation settings.


Identification of the high-molecular-mass mitochondrial oxaloacetate keto-enol tautomerase as inactive aconitase.
The homogeneous bovine heart mitochondrial high-molecular-mass oxaloacetate keto-enol tautomerase [(1988) Biochim. Biophys. Acta 936, 10-19] is shown to be an iron-sulfur protein as revealed by the enzyme spectral properties and direct chemical determination of non-heme iron and acid-labile sulfur. The protein is capable of catalysing the aconitase reaction after treatment with ferrous ions under anaerobic conditions. Treatment of the 'activated' protein with N-ethylmaleimide results in the simultaneous irreversible loss of the oxaloacetate keto-enol tautomerase and aconitase activities. The effects of some substrates and inhibitors on both activities show that the same catalytic site is involved in the oxaloacetate tautomerase and aconitase reactions. It is concluded that the protein previously described as a 80 kDa oxaloacetate keto-enol tautomerase is inactive aconitase.


Correlation between Maternal Characteristics during Early Pregnancy, Fetal Growth Rate and Newborn Weight in Healthy Pregnancies.
The objective of this study was to evaluate the association between maternal characteristics in early pregnancy and fetal growth (FG) and birth weight (BW). A prospective cohort study was performed in unselected pregnant women who attended an ultrasound evaluation at 11-14 weeks of pregnancy. Medical history, biochemical blood tests, biophysical variables and fetal weight at 20-25 and 30-36 weeks as well as the BW were assessed. Bivariate and multivariate linear models were constructed. In all, 543 patients with normal pregnancy and labor were selected. The multiple regression analysis showed a statistically significant association between maternal body mass index (BMI) in early pregnancy and the uterine artery pulsatility index (UtAPI) in the first trimester with BW (p < 0.0008) and with the ratio of fetal growth between the second and third trimesters (p < 0.0001). No correlation was found between these variables and first trimester levels of hemoglobin or glycemia. Maternal first trimester BMI and UtAPI correlate with the rate of intrauterine FG and with the BW. This evidence highlights the influence of maternal first trimester variables on fetuses with normal growth and the potential role of these variables in fetal programming.


A study of the serum 3,5,3'-triiodothyronine sulfate concentration in normal and hypothyroid fetuses at various gestational stages.
We have studied T3 sulfate (T3S) levels, blindly, in coded plasma samples from 21 normal and 3 hypothyroid fetuses at different stages of gestation (19-42 weeks). Fetal plasma samples were obtained by cordocentesis. T3S was detectable in all samples studied, with values ranging from 50-294 (mean +/- SD, 130 +/- 62 pmol/L). Plasma T3S was low (< 45 pmol/L) in all 4 normal adult control subjects studied simultaneously; serum T3S ranged from less than 20 to 130 in another set of 18 control subjects (mean +/- SD, 63 +/- 32 pmol/L). Fetal T3S values were positively correlated with gestational age (r = 0.43; P < 0.05), but not with free T4 (FT4), FT3, or TSH values. In the 3 hypothyroid fetuses at 31, 38, and 40 weeks gestation, respectively, plasma TSH was elevated (26, 98, and 24 mU/L, respectively), FT4 was low (10, 6.7, and 7.5 pmol/L, respectively), and FT3 was normal or high (3.2, 8.2, and 2.2 pmol/L, respectively). However, T3S values in hypothyroid fetuses (88, 133, and 252 pmol/L, respectively) were similar to those in normal fetuses at corresponding gestational ages. We conclude that 1) T3S is detectable in fetal circulation from at least 19 weeks gestation, and its concentration increases with fetal-age; 2) plasma T3S concentrations in the fetus at 19-40 weeks gestation are at least comparable to but generally higher than those in the adult; and 3) plasma T3S levels in hypothyroid fetuses are similar to those in normal fetuses. Recent studies demonstrating the ability of some fetal rat tissues (e.g. cerebral cortex) to desulfate T3S to T3 have suggested a possible role of T3S as a source of T3. Normal T3S in fetal hypothyroidism suggests that T3S may contribute to attenuation of the effects of hypothyroidism during intrauterine life.


The recoveryELISA--a newly developed immunoassay for measurement of therapeutic antibodies and the target antigen during antibody therapy.
Therapeutic antibodies are an important part of Biopharmaceuticals. They are highly innovative and specific drugs. Additionally, they play a challenging role in new demands in diagnostics because they disturb conventional antibody-based tests, such as immunoassays. The recoveryELISA is a newly developed immunoassay technology for monitoring such therapeutic antibodies or comparable biologics during the therapy. The recoveryELISA determines three results in one test: the free level of antigen (if available in serum), the level of therapeutic antibody and the specific dose-response interaction. The free level of antigen is the amount that can be measured in the immunoassay, as it exists unmasked under assay conditions. The relationship between therapeutic antibody level and neutralization rate of target protein is shown by the so-called 'recovery curve'. The recoveryELISA is demonstrated with the example of Omalizumab/IgE.


[Giant haemangioma of the liver - clinical and diagnostic problems - a case report].
Typical echodense haemangiomas of the liver can easily be identified by B-scan ultrasound. Atypical or giant haemangiomas of the liver, however, need to be differentiated from secondary tumours or primary liver carcinomas and cholangiocarcinomas. Until recently, computed tomography or magnetic resonance tomography were required to confirm the diagnosis, and not so rarely even a fairly risky fine-needle puncture had to be performed. Contrast media-assisted sonography, in general, offers the same scope of differentiating liver tumours as the radiographic tomography methods. Contrast media-assisted sonography is not yet widely available and has not been sufficiently evaluated. One important limitation is thought to be the lack of echo depth. We demonstrate the diagnostic value of this method in the case of a liver haemangioma measuring more than 20 cm in diameter. The resulting clinical questions are being discussed.


Genetic variability of hepatitis B virus isolates in Poland.
There is very limited knowledge about the genetic variability of HBV strains circulating in the population of Polish chronically infected HBV patients. The aim of this study was to analyse the phylogenetic relatedness and polymorphism in some functional domains of HBV genome among chronically infected patients from northern Poland. Fifty-one serum samples were included to analysis of HBV genomes due to the viral load sufficient for DNA preparation and sequencing. The sequences of the rt polymerase/S and preC/BCP regions of those isolates were analysed, compared to genome sequences of different variants of HBV from GenBank database and genetic relatedness of Polish genotypes to known reference strains was estimated. A phylogenetic tree of 41 analysed genotype A isolates as well as 8 genotype D strains was constructed showing relationship to know reference strains. Two isolates, initially classified as genotype F turned to be related to genotype H, newly described genotype deriving from genotype F, a very rare genotype in Europe. HBV genotypes' distribution pattern in Poland and phylogenetic relatedness seems to be different from our Eastern neighbours. Due to the fact that Poland is still ethnically uniform country, it is interesting to explore molecular epidemiology of HBV infections in our population.


Ossification sequence in infants who die during the perinatal period: population-based references.
To determine population-based references for the relationships between the presence of ossification centers and gestational age and skeletal length measurements among infants who die during the perinatal period, as well as to evaluate the possible influence of intrauterine growth restriction on ossification stage. During an 11-year period, nearly all infants who died perinatally in a well-defined geographic area routinely underwent radiography with a standardized technique. The presence of visible secondary ossification centers in the singletons (n = 495) was evaluated. Cluster analysis was used to identify stages of ossification; a sequential appearance of secondary ossification centers was assumed. Comparisons were made with Wilks lambda between male and female infants and between infants who were presumed to have growth restriction and those who were not. Reference ranges for the presence of ossification centers were calculated for interquartile ranges of femur length and gestational age. Eight clusters of ossification defining different stages of ossification of the pelvis, hindfeet, and knees were identified. The sequential clusters outlined well-defined intervals of femur length and gestational age. Bone lengths, birth weight, and gestational age within ossification clusters did not differ between the sexes (Wilks lambda = 0.989, P =.532) or according to whether growth restriction was presumed to exist (Wilks lambda = 0.958, P =.481). The reference diagrams calculated with this method indicate relationships between ossification sequence and both gestational age and skeletal length measurements.


Assessing genome-wide copy number aberrations and copy-neutral loss-of-heterozygosity as best practice: An evidence-based review from the Cancer Genomics Consortium working group for plasma cell disorders.
Plasma cell neoplasms (PCNs) encompass a spectrum of disorders including monoclonal gammopathy of undetermined significance, smoldering myeloma, plasma cell myeloma, and plasma cell leukemia. Molecular subtypes have been defined by recurrent cytogenetic abnormalities and somatic mutations that are prognostic and predictive. Karyotype and fluorescence in situ hybridization (FISH) have historically been used to guide management; however, new technologies and markers raise the need to reassess current testing algorithms. We convened a panel of representatives from international clinical laboratories to capture current state-of-the-art testing from published reports and to put forward recommendations for cytogenomic testing of plasma cell neoplasms. We reviewed 65 papers applying FISH, chromosomal microarray (CMA), next-generation sequencing, and gene expression profiling for plasma cell neoplasm diagnosis and prognosis. We also performed a survey of our peers to capture current laboratory practice employed outside our working group. Plasma cell enrichment is widely used prior to FISH testing, most commonly by magnetic bead selection. A variety of strategies for direct, short- and long-term cell culture are employed to ensure clonal representation for karyotyping. Testing of clinically-informative 1p/1q, del(13q) and del(17p) are common using karyotype, FISH and, increasingly, CMA testing. FISH for a variety of clinically-informative balanced IGH rearrangements is prevalent. Literature review found that CMA analysis can detect abnormalities in 85-100% of patients with PCNs; more specifically, in 5-53% (median 14%) of cases otherwise normal by FISH and cytogenetics. CMA results in plasma cell neoplasms are usually complex, with alteration counts ranging from 1 to 74 (median 10-20), primarily affecting loci not covered by FISH testing. Emerging biomarkers include structural alterations of MYC as well as somatic mutations of KRAS, NRAS, BRAF, and TP53. Together, these may be measured in a comprehensive manner by a combination of newer technologies including CMA and next-generation sequencing (NGS). Our survey suggests most laboratories have, or are soon to have, clinical CMA platforms, with a desire to move to NGS assays in the future. We present an overview of current practices in plasma cell neoplasm testing as well as an algorithm for integrated FISH and CMA testing to guide treatment of this disease.


[Persistent fever in the elderly].
We report the case of a 68 year old immuncompetent woman with persisting fever. Symptomatic acute CMV infection with a partial thrombosis of the left portal vein branch was diagnosed.


Solid-state nuclear magnetic resonance derived model for dynamics in the polypeptide backbone of the gramicidin A channel.
The dynamics of the backbone of the gramicidin A transmembrane cation channel in dimyristoylphosphatidylcholine bilayers have been investigated using solid state 15N nuclear magnetic resonance (n.m.r.) spectroscopy. With the temperature-dependent fluidity of the bilayer, the rates of motions in the helical gramicidin channel can be modulated. It is shown that in the gel phase, all substantial motions of the channel are slow on the timescale of the n.m.r. experiment (3.5 kHz). The use of oriented samples in which the axis of global channel rotation is aligned parallel to the magnetic field enables separation of global and local dynamics. Spectra obtained from oriented bilayer samples containing single-site 15N-labeled gramicidin at 8 degrees C are analyzed to yield a spatial model for local backbone motion. This model includes the axis of motion, the mean orientation, and the maximum amplitude of displacement for individual peptide planes. Specific sites in the first turn of the amino terminus were investigated, with emphasis on the Ala3 and Leu4 linkages, for which the orientation of the 15N chemical shift tensor with respect to the molecular frame has been determined. The effect of two well-characterized bilayer defect structures, parabolic focal conics and oily streaks, is included in the spectral simulations. It is found that only relatively small amplitude motions are possible at the two sites, with amplitudes of not more than +/- 8 degrees and +/- 15 degrees for the Ala3 and Leu4 sites, respectively. Detailed characterization of the bilayer surface geometry in the oriented samples is presently the major limiting factor in the use of this technique for probing the spatial extent of local motions in integral membrane proteins.


Fibroblast growth factor 21 reduces the severity of cerulein-induced pancreatitis in mice.
Fibroblast growth factor 21 (FGF21) acts as a hormonal regulator during fasting and is involved in lipid metabolism. Fgf21 gene expression is regulated by peroxisome proliferator-activated receptor (PPAR)-dependent pathways, which are enhanced during pancreatitis. Therefore, the aim of this study was to investigate FGF21's role in pancreatic injury. Fgf21 expression was quantified during cerulein-induced pancreatitis (CIP) or following mechanical or thapsigargin-induced stress through Northern blot analysis, in situ hybridization, and quantitative reverse transcription polymerase chain reaction. FGF21 protein was quantified by Western blot analysis. Isolated acinar cells or AR42J acinar cells were treated with recombinant FGF21 protein, and extracellular regulated kinase 1/2 activation was examined. The severity of CIP was compared between wild-type mice and mice overexpressing FGF21 (FGF21Tg) or harboring a targeted deletion of Fgf21 (Fgf21(-/-)). Acinar cell Fgf21 expression markedly increased during CIP and following injury in vitro. Purified FGF21 activated the extracellular regulated kinase 1/2 pathway in pancreatic acinar cells. The severity of CIP is inversely correlated to FGF21 expression because FGF21Tg mice exhibited decreased serum amylase and decreased pancreatic stellate cell activation, whereas Fgf21(-/-) mice had increased serum amylase and tissue damage. The expression of Fgf21 was also inversely correlated to expression of Early growth response 1, a proinflammatory and profibrotic transcription factor. These studies suggest a novel function for Fgf21 as an immediate response gene protecting pancreatic acini from overt damage.


Use of rigid and flexible bronchoscopy among pediatric otolaryngologists.
To explore how rigid and flexible bronchoscopy are used in pediatric otolaryngologic practice. Survey. Members of the American Society of Pediatric Otolaryngology who practice in the United States and Canada and were listed in the membership directory were eligible. Of the 206 members, 24 practicing outside the United States or Canada and 11 without an e-mail address or a fax machine were excluded. Hence, a questionnaire was e-mailed or faxed to 171 pediatric otolaryngologists. Questions concerned the practice setting, type and number of bronchoscopies, indications, complications, and medicolegal cases. Responses were received from 120 subjects (70.2%), with 3 retired and 2 practicing only otology, leaving 115 respondents who completed at least some of the questionnaire. Rigid and flexible bronchoscopy were performed by 72.7% (56/77) of those in academic settings and by 71.1% (27/38) of those in group or solo practices. In the last 12 months, approximately 10 454 total bronchoscopies were performed, with 2052 flexible and 9117 rigid bronchoscopies. Stridor, suspected foreign body inhalation, and laryngomalacia were the most common indications for bronchoscopy. Of the 83 respondents practicing rigid and flexible bronchoscopy, 25 (30.1%) used both instruments to manage complex or repeated foreign bodies, 25 (30.1%) used both to manage patients with cystic fibrosis, and 15 (18.1%) used both to manage simple foreign bodies. Complications were reported by 15.7% of the respondents, the most common being arrhythmia. Familiarity with a case resulting in medicolegal action was reported by 32.2% of the respondents. Rigid and flexible bronchoscopy have multiple uses in pediatric otolaryngologic practice. Also, flexible bronchoscopy appears to be emerging as a more frequently used diagnostic and therapeutic tool.


Does providing cost-effectiveness information change coverage priorities for citizens acting as social decision makers?
Escalating costs have generated increasing calls for Medicare to use cost-effectiveness as one criterion in determining coverage decisions. Decision-makers in U.S. health care have largely assumed that the public will reject any explicit consideration of cost in coverage policy, but there has been little formal testing of that hypothesis. We tested this assumption in a pilot study in which groups of citizens learned about and discussed health care costs, CEA methods, and common ethical issues embedded in CEA. Participants received information about 14 conditions and treatments and were asked to prioritize them for funding by Medicare under assumptions of constrained resources. Contrary to prevailing assumptions, this diverse sample understood CEA, were largely open to its use, and changed their own funding priorities when given cost-effectiveness ratio information.


Nucleotide sequence and revised map location of the arn gene from bacteriophage T4.
Non-glucosylated (Glu-) T-even phage DNAs are restricted by Escherichia coli RgIA and RgIB endonucleases with different specificities. RgIB endonuclease activity is strongly inhibited by anti-restriction endonuclease (Arn) encoded by the bacteriophage T4 genome. The nucleotide sequence of the arn gene encoding Arn was determined. The product of the cloned arn gene was overexpressed by the T7 RNA polymerase/promoter system, and its molecular size is consistent with that predicted from the open reading frame of the arn gene. The arn gene is located between the asiA gene and motA gene in the region of 161,300-161,578 nucleotides.


Development and validation of an algorithm to identify patients newly diagnosed with HIV infection from electronic health records.
An algorithm was developed that identifies patients with new diagnoses of HIV infection by the use of electronic health records. It was based on the sequence of HIV diagnostic tests, entry of ICD-9-CM diagnostic codes, and measurement of HIV-1 plasma RNA levels in persons undergoing HIV testing from 2006 to 2012 at four large urban Veterans Health Administration (VHA) facilities. Source data were obtained from the VHA National Corporate Data Warehouse. Chart review was done by a single trained abstractor to validate site-level data regarding new diagnoses. We identified 1,153 patients as having a positive HIV diagnostic test within the VHA. Of these, 57% were determined to have prior knowledge of their HIV status from testing at non-VHA facilities. An algorithm based on the sequence and results of available laboratory tests and ICD-9-CM entries identified new HIV diagnoses with a sensitivity of 83%, specificity of 86%, positive predictive value of 85%, and negative predictive value of 90%. There were no meaningful demographic or clinical differences between newly diagnosed patients who were correctly or incorrectly classified by the algorithm. We have validated a method to identify cases of new diagnosis of HIV infection in large administrative datasets. This method, which has a sensitivity of 83%, specificity of 86%, positive predictive value of 85%, and negative predictive value of 90% can be used in analyses of the epidemiology of newly diagnosed HIV infection.


Postoperative analgesia by epidural infusion of ropivacaine and fentanyl versus intravenous administration of morphine in patients undergoing radical retropubic prostatectomy: results of a prospective study.
Radical prostatectomy is followed by postoperative pain, that can be linked to potential morbidity. Aim of this study is to compare efficacy of epidural infusion of ropivacaine and fentanyl versus intravenous administration of morphine in terms of reduction of pain and possible complications. We have considered 73 consecutive patients who underwent a retropubic radical prostatectomy. For analgesia control we have considered the use of epidural infusion of ropivacaine and fentanyl (Group R) in 48 patients, compared to continuous intravenous administration of morphine (Group M) in 25 patients. Evaluation criteria have been: blood pressure, heart and breath rate, sedation state, nausea and vomit, entity of pain with VRS scale (Verbal Rating Score) at rest and in movement in the first 72 hours, canalization and need of additional analgesia. VRS values were lower in Group R especially during movement, and need of additional analgesia was reported in one patient of Group R, and in three patients of Group M; assumption of morphine was interrupted in three patients. Epidural catheter was removed for malfunction in three patients, and seven patients complained for leg paresthesia. Sedation resulted higher in group M. Mobilization in second day occurred with fewer difficulties for patients of group R, while canalization resulted delayed in group M. Results underlined a good analgesic cover in both groups. Advantages of epidural analgesia are represented by the use of smaller doses of opioids, fewer cardiocirculatory and respiratory side effects, lower incidence of nausea and vomit, early canalization and post-operative mobilization.


Porteus Maze Q score as a measure of impulsivity.
The validity of the Q score (derived from the qualitative scoring of the Porteus Maze Test) as a measure of impulsivity was investigated by comparing the performance of high- and low-impulse groups. 80 Ss were employed and the Q score failed to differentiate high- from low-impulse institutionalized Ss, delinquent from nondelinquent Ss and extreme groups based on ratings. These findings are at variance with the claim that the Q score measures impulsivity.


Postoperative ventilatory performance: dependence upon surgical incision.
Sixty-one patients undergoing elective surgery were evaluated in regard to the degree of depression in FVC and FEV1 that occurred in the postoperative period. Following a thoracotomy a reduction of about 75 per cent occurred, after a long midline incision 68 per cent, after an upper midline or paramedian incision 61 per cent, after a subcostal incision 53 per cent and after a lower midline incision 38 per cent. This information may be useful in determining which patients are at an increased risk for postoperative respiratory insufficiency.


[Abstracts of VIII Young Scientists Conference dedicated to 55th anniversary of Central Research Institute of Dentistry and Maxillofacial Surgery 'Modern scientific achievements in dentistry and maxillofacial surgery'].
Abstracts of VIII Young Scientists Conference dedicated to 55th anniversary of Central Research Institute of Dentistry and Maxillofacial Surgery 'Modern scientific achievements in dentistry and maxillofacial surgery'. Therapeutic stomatology. Surgical dentistry. Maxillofacial surgery. Orthopedic stomatology. Orthodontics. Organization of dental care.


Skeletal remineralization after surgery for primary and secondary hyperparathyroidism.
Bone mineral was measured by photon absorptiometry before and after parathyroid surgery in patients with primary or secondary hyperparathyroidism (HPT). The mean bone mineral density of lumbar vertebrae was 0.82 +/- 0.04 (SEM) gm/cm2 in primary HPT (n = 7) and 0.86 +/- 0.05 gm/cm2 in secondary HPT (n = 11). These values are significantly lower than for age-matched normal subjects. After successful parathyroid surgery, the bone mass of the distal radius and lumbar vertebrae increased by approximately 10% within 3 months after operation and then remained stable during the first postoperative year. In conclusion, parathyroid surgery is followed by a significant increase of bone mass in primary and secondary HPT. The substantial increase in bone mass in parts of the skeleton consisting predominantly of trabecular bone, as well as in sites with predominantly cortical bone, indicates that remineralization after operation involves a generalized increase in bone mass.


[Development of a computer-aided system for augmentation mammaplasty simulation].
A method for developing a three-dimensional visual system for simulating augmentation mammaplasty based on OpenGL is proposed. The 3D reconstruction of breast surface using NURBS, reality simulation of the breast surface, parametric design of mammary prosthesis, and simulation of postoperative effect are described. The system may provide a means for better communication between the surgeons and patients.


Health-related quality of life in haematopoietic stem cell transplant survivors: a qualitative study on the role of psychosocial variables and response shifts.
Psychosocial assessment prior to haematopoietic stem cell transplants (HSCT) can help to identify patients at risk of impaired health-related quality of life (HRQOL) post-transplant. According to the response-shift model, certain antecedents and mechanisms, along with changes in internal standards, values or conceptualizations of HRQOL, facilitate adjustment to changes in health circumstances. This study sought to explore the role of psychosocial variables in adjustment to compromised HRQOL following HSCT, from the theoretical basis of the response-shift model. Semi-structured interviews were conducted with 28 patients (15 women, 13 men; 22-71 years), post-HSCT. Time since transplant ranged from 1 month to 28 years. Verbatim transcripts were analysed using template analysis. Patients provided narrative examples of changing their values and internal standards. Optimism, social support, social comparisons, changing expectations and setting goals were identified as important in managing threats to HRQOL. The response-shift model is a useful theoretical basis for exploring HRQOL in HSCT patients. Response shifts and psychosocial variables may help patients to cope and enabling them to experience good HRQOL despite the negative effects of HSCT. Understanding the adjustment processes has implications for patient care.


The Addition of Intravenous Metronidazole to Oral Vancomycin is Associated With Improved Mortality in Critically Ill Patients With Clostridium difficile Infection.
The optimal therapy for critically ill patients with Clostridium difficile infection (CDI) is not known. We aimed to evaluate mortality among critically ill patients with CDI who received oral vancomycin (monotherapy) vs oral vancomycin with intravenous (IV) metronidazole (combination therapy). A single-center, retrospective, observational, comparative study was performed. Patients with a positive C. difficile assay who received oral vancomycin while bedded in an intensive care unit (ICU) between June 2007 and September 2012 were evaluated. Patients meeting ≥3 of the following criteria were included: albumin <2.5 g/dL, heart rate >90 bpm, mean arterial pressure <60 mmHg, white blood cell count ≥15 000 cells/mL, age >60 years, serum creatinine ≥1.5 times baseline, or temperature ≥100.4°F. Patients in the combination therapy group received IV metronidazole within 48 hours after initiating vancomycin. Patients <18 years or with unrelated gastrointestinal disease were excluded. The primary outcome was in-hospital mortality. Patients were matched using Acute Physiology and Chronic Health Evaluation II scores. Eighty-eight patients were included, 44 in each group. Patient characteristics were similar although more patients in the combination group had renal disease. Mortality was 36.4% and 15.9% in the monotherapy and combination therapy groups, respectively (P = .03). Secondary outcomes of clinical success, length of stay, and length of ICU stay did not differ between groups. Our data are supportive of the use of combination therapy with oral vancomycin and IV metronidazole in critically ill patients with CDI. However, prospective, randomized studies are required to define optimal treatment regimens in this limited population of CDI patients.


Treating Addiction to Prescription Opioids.
For over 30 years, medication-assisted treatment for preventing relapse in opiate use disorder has been limited to methadone maintenance and withdrawal treatments followed by naltrexone; however, buprenorphine has emerged as an exciting and successful pharmacotherapy alternative. Buprenorphine is a partial μ-opioid receptor agonist with less abuse potential and a favorable safety profile compared to methadone and offers a great opportunity of office-based opioid addiction treatment (OBOT) to improve patient access. However, we have limited number of providers certified to prescribe buprenorphine in South Dakota. We need to address potential barriers for its expansion, including concerns of diversion and abuse, insufficient knowledge on buprenorphine and addiction treatment, and lack of access to addiction specialists and treatment resources. Since buprenorphine does retain some abuse potential, good office practice needs to be implemented as we expand its access. We need to choose appropriate candidates for OBOT with clear expectations at the intake. We need to enforce urine drug screens, pill counts, and use of prescription monitoring programs. It is also important to provide or refer patients to addiction counseling services and more intensive levels of care including methadone maintenance and residential care when needed.


[Martian environment and life].
Five conditions for life to arise are discussed with referring to early Martian environment. The key to determine whether any life form appeared on Mars is found to be the early Martian carbon dioxide atmospheric pressure and then the temperature. The importance to determine the heat flow is indicated. The items to be measured for future Martian exploration are listed. The surface materials, which has been poorly understood, are emphasized for further exploration. Two strategies for search for life on Mars, "step by step" strategy and quick strategy, are suggested.


Bioanalytical method development and validation of natamycin in rabbit tears and its application to ocular pharmacokinetic studies.
A new selective and sensitive high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of natamycin in rabbit tears using amphotericin B as internal standard (IS). Chromatographic separation was achieved on a Luna Cyano column (100 mm × 2 mm, 3 μm) using ammonium acetate buffer (pH 4; 3.5mM): methanol (10:90, v/v) as the mobile phase. The run time was 5 min. Detection was performed by negative ion electrospray ionization in multiple reaction monitoring (MRM) mode. The calibration curve was linear over the concentration range from 25 to 800 ng/ml, and lower limit of detection of 12.5 ng/ml. The accuracy and precision of the method were within the acceptable limit of ± 20% at the lower limit of quantitation and ± 15% at other concentrations. Natamycin was stable during the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and 30 days storage in a freezer at -70 ± 10 °C. The method was successfully applied to the ocular pharmacokinetic studies of natamycin eye drops in New Zealand rabbit tears.


Efficient purification of small unsaturated oligoglucuronides by reversed-phase chromatography.
Ion-exchange chromatography has been applied to purification of unsaturated oligoglucuronans. After an isocratic elution on a strong anion-exchange column, the collected fractions were desalted by low pressure size exclusion chromatography. However, this efficient separation was limited by the time required to desalt. So, we developed a reversed-phase chromatography method using back ionization of oligomers. Two C18 columns were tested with trifluoroacetic acid (TFA 0.7%) as eluent. Different selectivities and column stabilities were observed in this acidic condition. The scale up for semi-preparative applications enabled us to recover pure unsaturated oligoglucuronans without desalting step.


Concentrating membrane proteins using asymmetric traps and AC electric fields.
Membrane proteins are key components of the plasma membrane and are responsible for control of chemical ionic gradients, metabolite and nutrient transfer, and signal transduction between the interior of cells and the external environment. Of the genes in the human genome, 30% code for membrane proteins (Krogh et al. J. Mol. Biol.2001, 305, 567). Furthermore, many FDA-approved drugs target such proteins (Overington et al. Nat. Rev. Drug Discovery 2006, 5, 993). However, the structure-function relationships of these are notably sparse because of difficulties in their purification and handling outside of their membranous environment. Methods that permit the manipulation of membrane components while they are still in the membrane would find widespread application in separation, purification, and eventual structure-function determination of these species (Poo et al. Nature 1977, 265, 602). Here we show that asymmetrically patterned supported lipid bilayers in combination with AC electric fields can lead to efficient manipulation of charged components. We demonstrate the concentration and trapping of such components through the use of a "nested trap" and show that this method is capable of yielding an approximately 30-fold increase in the average protein concentration. Upon removal of the field, the material remains trapped for several hours as a result of topographically restricted diffusion. Our results indicate that this method can be used for concentrating and trapping charged membrane components while they are still within their membranous environment. We anticipate that our approach could find widespread application in the manipulation and study of membrane proteins.


Endoscopic endonasal laser versus endonasal surgical dacryocystorhinostomy for epiphora due to nasolacrimal duct obstruction: prospective, randomised, controlled trial.
Rhinostomy patency is a problem in all forms of dacryocystorhinostomy. Laser-assisted procedures are potentially fast and result in excellent haemostasis. However, they may induce more fibroblastic activity, resulting in excessive scarring and stenosis of the rhinostomy, compared with non-laser dissection. The objective of this study was to compare subjective outcomes following dacryocystorhinostomy conducted with endoscopic endonasal laser and with endonasal surgical techniques. Prospective, randomised, controlled trial comparing potassium titanyl phosphate endonasal laser dissection with endonasal surgical techniques, for dacryocystorhinostomy to treat epiphora due to primary, acquired nasolacrimal duct obstruction. One hundred and twenty-six adult patients with chronic epiphora due to primary, acquired nasolacrimal sac or duct obstruction. INTERVENTIONS comprised endonasal dacryocystorhinostomy, performed using potassium titanyl phosphate laser or surgical dissection, in order to open the lacrimal sac into the nasal cavity. These were: symptom score and symptom relief of epiphora (at three and 12 months post-procedure); duration and ease of procedure; and duration of hospital stay. Sixty patients underwent endonasal laser dacryocystorhinostomy and 66 underwent endonasal surgical dacryocystorhinostomy. Symptomatic success was 82 per cent at three months and 68 per cent at 12 months in the laser group, and 76 per cent at three months and 74 per cent at 12 months in the surgical group. A two-point reduction in the symptom score was associated with symptomatic success at three and 12 months. Eight patients randomised to the laser group required additional instrumentation in order to remove thick bone over the lacrimal sac. At three months, endonasal laser dacryocystorhinostomy had better results than endonasal surgical dacryocystorhinostomy. However, at 12 months, the surgical procedure had better results than the laser procedure. There was no statistically significant difference between the two groups at three or 12 months with regard to symptomatic outcome. The ease of procedure (on a scale of zero to 10) was 4.5 for the laser procedure and 4.1 for the surgical procedure. The average times for the procedures were 25 minutes in the laser group and 20 minutes in the surgical group. No statistical difference was found when comparing: symptom score improvement for local anaesthetic vs general anaesthetic; ages over and under 70 years; laterality; or operating surgeon. Change in the symptom score was a useful indicator of symptomatic success.


An organocatalytic approach to enantiomerically enriched α-arylcyclohexenones and cyclohexanones.
The presence of a p-nitrophenyl group converts acetone into an excellent and versatile nucleophile in organocatalytic processes, able to react with α,β-unsaturated aldehydes affording β-substituted α-arylcyclohexenones via a Michael reaction/aldol reaction/dehydration sequence, which occurs in good yields, ee up to 96% and complete diastereoselectivity. The resulting compounds are excellent synthons for the diastereoselective preparation of a variety of synthetically useful polysubstituted cyclohexanones and derivatives.


[DNA diagnosis of hemophilia A in a family without an affected proband].
A pedigree is described where the proband affected by hemophilia A, as well as his healthy brother, have died (so that their DNA was not available for analysis), and yet, the determination of five linked RFLPs in the remaining pedigree members made it possible to ascertain the carrier status in the proband's grand-daughter. (Fig. 1, Tab. 1, Ref. 16).


Free serosal cells originating from the embryo of the wasp Diadromus pulchellus in the pupal body of parasitized leek-moth, Acrolepiosis assectella. Are these cells teratocyte-like?
In braconid species, teratocytes are derived from a serosal cell membrane which envelops the developing parasitoid embryo. On hatching, this membrane dissociates into individual cells, the teratocytes, which then circulate in the haemolymph of the host. We describe herein such a membrane, surrounding the embryo in eggs of the ichneumonid parasitoid wasp, Diadromus pulchellus. This membrane consisted of a single sheet of tightly packed cells with large 12+/-1.4 &mgr;m nuclei. These cells were released after hatching in vitro and cells of the same size were detected in vivo, in the vicinity of the D. pulchellus embryo. The number of nuclei detected suggests that the serosal membrane consists of about 450+/-150 cells. These cells did not grow after hatching of the parasitoid egg in the parasitized host, Acrolepiosis assectella, during the development of the parasitoid wasp larva. Southern blot experiments, using D. pulchellus satellite DNA or the ribosomal genes as probes, showed that free-living floating cells of wasp origin were present in the body of the parasitized host. This is the first time that free-floating teratocyte-like cells have been described in species of the Ichneumonidae.


Open vessel and cooling while heating microwave-assisted synthesis of pyridinyl N-aryl hydrazones.
We reported the first example of open vessel and cooling while heating microwave-assisted synthesis of pyridinyl N-aryl hydrazones. Compounds were prepared in excellent isolated yields (88-98%) in only 5 min, by reacting 4- and 2,4-(di)substituted phenylhydrazines, bearing both electron-donating (4-CH₃, 4-OCH₃) and -withdrawing (4-Cl, 4-Br, 4-CF₃, 4-NO₂, 2,4-Cl₂) groups with 2-, 3-, and 4-acetylpyridine. The method was successfully extended to other carbonyl compounds.


[Screening strategies for vulvar preneoplastic and neoplastic lesions].
Although they cover superficial areas, preneoplastic and neoplastic lesions of the vulva are often diagnosed late. The reasons for this delay is the low incidence of this invasive neoplasm, the advanced age of patients, the non-specific symptoms mainly taking the form of itch, burning, dyspareunia and blood loss, which are also compatible with a non-neoplastic infective pathology. The late diagnosis of carcinoma of the vulva may also be linked to the inadequate examination of the external genitals by doctors as a result of insufficient specific knowledge. Epidemiological data and the natural history of VIN lesions and carcinoma of the vulva argue that mass screening is not feasible, but an adequate programme of early diagnosis must be introduced. Early diagnosis is linked to three key elements: targeted anamnesis, clinical examination and the appropriate use of the various diagnostic procedures. Vulvoscopy represents the most reliable method, above all because it allows a biopsy to be taken of any suspected lesion. From January 1992 to December 1998, a total of 1678 vulvoscopies were performed at the Institute of Clinical Obstetrics and Gynecology of the University of Catania in patients aged between 16 and 82 years old. Biopsies were taken of all suspected lesions. Sixty-nine cases of VIN (4.11%) were diagnosed: 28 VIN1, 24 VIN2 and 17 VIN3. Lesions were only symptomatic in 39.1% of cases. The association of vulvoscopy with biopsy of suspected lesions, even in the absence of vulvar symptoms, represents the most efficacious method for the diagnosis of intraepithelial lesions.