Mechanisms of respiration-induced changes in vasomotor control exerted by rostral ventrolateral medulla.
Efferent and afferent mechanisms involved in the differential depressor response to inactivating rostral ventrolateral medulla (RVLM) under conditions of normal (2.5 ml) and reduced (1.5 ml) tidal volume were examined in urethan-anesthetized rats. Under conditions of reduced tidal volume, bilateral microinjection of lidocaine (200 nl, 4%) into RVLM produced an attenuated fall in both mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA) despite the fact that base-line levels of both are unchanged. The specificity of the differential response was tested by examination of regional hemodynamics. Microinjecting lidocaine into RVLM produced an attenuated fall in renal, mesenteric, and hindquarter vascular resistances in parallel with the attenuated fall in MAP produced by reducing tidal volume. Microelectrode stimulation of RVLM produced an increase in resistance in all three vascular beds. Of the neural sensory systems examined, both vagal and chest wall afferents appear to be involved in mediating the differential depressor response. These results demonstrate the importance of afferent sensory input to the medulla in determining sites of generation and maintenance of sympathetic vasomotor tone.


Social Information Processing in Anger Expression and Partner Violence in Returning U.S. Veterans.
We examined social information processing factors that could represent pathways through which posttraumatic stress disorder (PTSD) symptoms relate to anger expression and intimate partner violence (IPV) perpetration in returning U.S. veterans. The sample included 92 male Operation Enduring Freedom/Operation Iraqi Freedom veterans, primarily Caucasian (77.4%), with smaller numbers of African American, Asian, Hispanic or Latino, American Indian or Alaskan Native, and other minority participants (9.7%, 2.2%, 2.2%, 3.2%, and 5.3% respectively). The average age was 40.37 (SD = 9.63) years. Data were collected through self-report questionnaires (PTSD Checklist, State-Trait Anger Expression Scale, Revised Conflict Tactics Scales) and the Articulated Thoughts in Simulated Situations experimental protocol. Laboratory-based assessment of cognitive biases and hostile attributions were tested as mediators of associations between PTSD symptoms and anger expression and IPV. Among the PTSD symptom clusters, hyperarousal symptoms were most strongly associated with anger expression (r = .50) and IPV perpetration (r = .27). Cognitive biases mediated associations between PTSD total scores and 3 of 4 PTSD cluster scores as well as anger expression. Hostile attribution biases were also associated with IPV perpetration (r = .23). We discuss the implications of these findings for understanding social information processing mechanisms for the relationship between PTSD symptoms and aggression.


C1qTNF-related protein-6 protects against doxorubicin-induced cardiac injury.
The clinical use of doxorubicin (DOX) is limited by its toxic effect. However, there is no specific drug that can prevent DOX-related cardiac injury. C1qTNF-related protein-6 (CTRP6) is a newly identified adiponectin paralog with many protective functions on metabolism and cardiovascular diseases. However, little is known about the effect of CTRP6 on DOX-induced cardiac injury. The present study aimed to investigate whether CTRP6 could protect against DOX-related cardiotoxicity. To induce acute cardiotoxicity, the mice were intraperitoneally injected with a single dose of DOX (15 mg/kg). Cardiomyocyte-specific CTRP6 overexpression was achieved using an adenoassociated virus system at 4 weeks before DOX injection. The data in our study demonstrated that CTRP6 messenger RNA and protein expression were decreased in DOX-treated hearts. CTRP6 attenuated cardiac atrophy induced by DOX injection and inhibited cardiac apoptosis and improved cardiac function in vivo. CTRP6 also promoted the activation of protein kinase B (AKT/PKB) signaling pathway in DOX-treated mice. CTRP6 prevented cardiomyocytes from DOX-induced apoptosis and activated the AKT pathway in vitro. CTRP6 lost its protection against DOX-induced cardiac injury in mice with AKT inhibition. In conclusion, CTRP6 protected the heart from DOX-cardiotoxicity and improves cardiac function via activation of the AKT signaling pathway.


Polymorphisms in the human cysteine-rich secretory protein 2 (CRISP2) gene in Australian men.
Cysteine-rich secretory protein 2 (CRISP2) is localized to the human sperm acrosome and tail. It can regulate ryanodine receptors Ca(2+) gating and binds to mitogen-activated protein kinase kinase kinase 11 in the acrosome and gametogenetin 1 (GGN1) in the tail. In order to test the hypothesis that CRISP2 variations contribute to male infertility, we screened coding and flanking intronic regions in 92 infertile men with asthenozoo- and/or teratozoospermia and 176 control men using denaturing HPLC and sequencing. There were 21 polymorphisms identified, including 13 unreported variations. Three SNPs resulted in amino acid substitutions: L59V, M176I and C196R. All were only present in a heterozygous state and found in fertile men. However, the C196R polymorphism was of particular interest as it resulted in the loss of a strictly conserved cysteine involved in intramolecular disulphide bonding. Screening of an additional 637 infertile men identified 23 heterozygous C196R men to give an overall frequency of 3.6%, compared with 3.4% in control men. The functional significance of the C196R polymorphism was defined using a yeast two-hybrid assay. The C196R substitution resulted in the loss of CRISP2-GGN1 binding. Although none of the many polymorphisms identified herein showed a significant association with male infertility, functional studies suggested that the C196R polymorphism may compromise CRISP2 function.


A quantitative analysis of the effect of all-trans-retinoic acid on the pattern of chick wing development.
Small, positively charged beads that slowly release known amounts of all-trans-retinoic acid have been implanted below the apical ectodermal ridge at the anterior margin (opposite somite 16) of wing buds of 3 1/2 day-old chick embryos. The continuous release of retinoic acid is shown to create an anteroposterior concentration gradient of retinoic acid in the limb field that is stable with time, despite the fact that this compound is metabolized by the limb tissue. With beads that release increasing amounts of retinoic acid, the normal 234 digit pattern is progressively altered to a 2234, to a 32234, and then to a 432234 pattern. The tissue concentrations of all-trans-retinoic acid required to change the digit pattern in this way range between 1 and 25 nM. When the same amounts of retinoic acid are released from posteriorly implanted beads (placed below the apical ectodermal ridge opposite somite border 19/20 or somite 20), the normal digit pattern is unaffected. Implantations of beads that release all-trans-retinoic acid are thus identical in their effect to grafts of cells from the limb polarizing region, which cause similar dose-dependent changes in the digit pattern when grafted to the anterior margin of the bud (but not when grafted opposite somites 19 or 20). Because of the low concentrations of retinoic acid required for its biological effect, the graded response observed, and the fact that a concentration gradient is established across the limb field, all-trans-retinoic acid closely mimics the putative morphogen that has been postulated to be emitted by polarizing region cells during normal development.


Psychiatrists' religious attitudes in relation to their clinical practice: a survey of 231 psychiatrists.
A total of 231 psychiatrists working in several London teaching hospitals were surveyed about their private religious attitudes and how these affected their reported clinical practice. Although only 27% reported a religious affiliation and 23% a belief in God, 92% felt that psychiatrists should concern themselves with the religious concerns of their patients. Although the psychiatrists who were religious were more likely to make referrals to religious leaders or to disclose their own beliefs to patients, contrary to reports from the United States there was no evidence that psychiatrists' private religious beliefs had an important influence on their clinical practice.


[Psychopathological characteristics and social functioning of the schizophrenic patients with the T allele of the MTHFR677C>T polymorphism].
To compare the severity of psychopathological symptoms and characteristics of personal and social functioning in patients with schizophrenia, carriers of different alleles of the MTHFR677C>T polymorphism. One hundred and fifty patients diagnosed with schizophrenia were genotyped for the MTHFR677C>T polymorphism, some of them were examined using psychometric scales and tests (PANSS, SANS, PSP, CDSS, a battery of cognitive tests etc). Patients with the MTHFR677T variant had greater severity of negative symptoms regardless of psychometric instruments used in the study, greater severity of schizophrenia disorder in whole measured with the PANSS General psychopathological symptoms subscale, higher scores on the PANSS cognitive cluster and lower levels of personal and social functioning measured with the PSP compared to the patients with the MTHFR677CС genotype. The results of psychometric testing are in line with significant differences of MTHFR677T allele carriers from those with the MTHFR677CС genotype in the number of disable patients and the use of different types of psychiatric services. The differences were not related to sex, age, illness duration and depressive symptoms measured with CDSS and PANSS «depression/anxiety» factor.


Unraveling sorption of lead in aqueous solutions by chemically modified biochar derived from coconut fiber: A microscopic and spectroscopic investigation.
In this study, we examined the efficacy of nine different types of coconut-fiber derived biochars (CFBs), prepared at different temperatures and chemically modified with ammonia, hydrogen peroxide and nitric acid, to remove lead (Pb2+) from aqueous solutions. Langmuir-qm values of the biochars pyrolyzed at 300°C and modified with ammonia and nitric acid increased from 49.5 to 105.5 and 85.2mgg-1, respectively, compared to control (unmodified), whereas hydrogen peroxide treatment had no effect. The maximum amount of Pb adsorbed on biochars was in the order of CFB-700>MCFB-300-NH3·H2O>CFB-500>MCFB-300-HNO3>CFB-300. X-ray absorption fine structure (XAFS) spectroscopy results revealed that Pb-montmorillonite, Pb(C2H3O2)2, PbSO4, Pb-Al2O3 and Pb3(PO4)2 were the five most important Pb species observed in Pb-loaded biochars, and as such, favoring Pb immobilization in aqueous solutions. Overall, the sorption capacity of CFBs pyrolyzed at 300°C substantially increased for Pb2+ with ammonia and nitric acid modification. However, these chemical modifications did not improve the sorption of Pb on CFBs pyrolyzed at temperatures ≥500°C, thereby highlighting a temperature dependent response of chemically modified biochars to Pb sorption in this study.


Morphometric estimation of pulmonary diffusion capacity. VII. The normal Guinea pig lung.
A study of 15 normal adult guinea pig lungs fixed by instillation of fixatives via the airways is described. The morphological appearance of the guinea pig lung is similar to that of other mammals apart from quantitative differences in the mass of connective tissue in the alveolar-capillary barrier whose mean thickness was estimated at 1.59 mum. Lung volume, alveolar volume, capillary volume and the surface areas of alveoli and capillaries are shown to be linearly related to body weight. Analysis of the morphometric diffusion capacities of the alveolar-capillary membrane and of the lung shows that these are also linearly related to body weight. For a mean body weight of 429 g the lung volume was 13 ml, the alveolar surface 0.91 m2, the capillary volume 1.5 ml, the harmonic mean barrier thickness 0.42 mum, and DL 2.13 ml/min.mm Hg. These parameters are compared with those of other mammalian species and discussed in the context of normal activity levels as a probable important adaptive factor determining the size of the gas exchange apparatus.


Lysosomal and ATP changes after renal-stalk clamping in rats. I. Protective action of Trasylol.
The action of Trasylol on the lysosomal and ATP changes after renal-stalk clamping in rats was examined. It was found that, in the presence of an effective Trasylol concentration, a distinct stabilization of the lysosomal membrane can be detected after 30 min of renal-stalk clamping. It was also found that there is only an indirect relationship between lysosomal changes and ATP metabolism under the action of Trasylol. The more rapid regeneration of the ATP is a result of the better microcirculation. The results confirm the earlier belief that Trasylol acts primarily on the lysosomal membrane.


Peptide-TiO2 surface interaction in solution by ab initio and molecular dynamics simulations.
Ab initio periodic calculations and classical molecular dynamics (MD) simulations were performed to investigate the adsorption mode of alanine and a number of short peptides, in particular two peptides, alanine-glutamic acid and alanine-lysine, taken as model systems for the ionic self-complementary oligopeptide EAK16-II, onto TiO(2) (110) rutile surface, and their conformational characteristics upon adsorption. The atomistic description of the rutile surface and its interactions with water and peptide molecules were based on ab initio calculations, the TIP3P water model, the AMBER force field, and available parameters. By comparison with ab initio calculations, it is shown that MD simulations of reasonable duration can describe the main characteristics of the peptide-TiO(2) surface interaction in solution, at least on a short time scale. Atom-atom radial distribution functions, atom-surface distances, backbone and side chain dihedral angle distributions, and peptide-surface interaction energies have been analyzed. Once adsorbed onto the TiO(2) rutile surface by a bidentate interaction of both carboxyl oxygens with two adjacent Ti atoms, the small peptide studied showed a clear propensity to remain there and undergo relatively limited hinge-bending motions.


Décalage in moral judgement as a measure of deactualisation weakness of schizophrenic adolescents.
12 schizophrenic, 11 borderline and 20 neurotic adolescents were examined with the procedure, according to Kohlberg, to moral development. In addition, existence techniques, themes and formal personality attributes according to Thomae were assessed. In schizophrenic adolescents a very distinct discrepancy between average and the highest stage values of the moral judgement were the main result (décalage value). This distinct décalage corresponds with the previously discussed distinct ambiguity tolerance of schizophrenics that comes into being on the basis of a deactualisation weakness towards situatively stimulated representative pieces of existence. In connection with this décalage and with the other values for moral judgement are findings on life style and theme that can be interpreted in the sense of an introversion and a cry for help connected with increased resistance and present a form of illness mastering. A detachment from the parents only appears possible at the cost of an increased lability (in the area of moral and ego-identity as décalage).


Effects of calcium gluconate on experimental periodontitis and alveolar bone loss in rats.
We examined the effects of calcium gluconate, an anti-inflammatory calcium salt, on ligature-induced experimental periodontitis and related alveolar bone loss. Calcium gluconate was orally administered daily for 10 days at 250, 125 or 62.5 mg/kg, beginning 1 day after ligation. We recorded changes in body-weight and alveolar bone loss and quantified the anti-inflammatory effects of calcium gluconate by measuring levels of myeloperoxidase (MPO), IL-1β and TNF-α. We also evaluated inducible nitric oxide synthase (iNOS) activity and malondialdehyde (MDA) concentration as a measure of antioxidant effects. Ligature placement produced a marked decrease in body-weight, increased alveolar bone loss, and led to increased MPO, IL-1β, TNF-α and MDA concentrations, as well as elevated iNOS activity, increased inflammatory cell infiltration and decreased collagen fibre content in gingival tissue. Histopathology revealed decreased alveolar bone volume, increased osteoclast cell numbers and activity, and an elevated percentage of osteclasts on the alveolar bone surface. The effects of ligature placement were significantly and dose-dependently inhibited by 10 days of daily oral treatment with 250 and 125 mg/kg of calcium gluconate. The results suggest that 10 days daily oral treatment with calcium gluconate effectively inhibits ligature placement-induced periodontitis and related alveolar bone loss via antioxidant effects.


Controllable Synthesis of Highly Luminescent Boron Nitride Quantum Dots.
Boron nitride quantum dots (BNQDs), as a new member of heavy metal-free quantum dots, have aroused great interest in fundamental research and practical application due to their unique physical/chemical properties. However, it is still a challenge to controllably synthesize high-quality BNQDs with high quantum yield (QY), uniform size and strong fluorescent. In this work, BNQDs have been successfully fabricated by the liquid exfoliation and the subsequent solvothermal process with respect to its facileness and easy large scale up. Importantly, BNQDs with high-quality can be controllably obtained by adjusting the synthetic parameters involved in the solvothermal process including filling factor, synthesis temperature, and duration time. Encouragingly, the as-prepared BNQDs possess strong blue luminescence with QY as high as 19.5%, which can be attributed to the synergetic effect of size, surface chemistry and edge defects. In addition, this strategy presented here provides a new reference for the controllable synthesis of other heavy metal-free QDs. Furthermore, the as-prepared BNQDs are non-toxic to cells and exhibit nanosecond-scaled lifetimes, suggesting they have great potential biological and optoelectronic applications.


Effects of nicotine and a cannabinoid receptor agonist on negative contrast: distinction between anxiety and disappointment?
Animals trained to lick for a sucrose solution of a given incentive value that subsequently encounter an incentive downshift (i.e. 32-4% sucrose) display an exaggerated decrease in the amount consumed, relative to unshifted controls. This change has been classified as a successive negative contrast (SNC) effect. The emotional component to this robust behavioural change is dynamic and changes from post-shift day (PSD) 1 to 2. Anxiolytics block SNC, but the possible link between anxiety and SNC needs further exploration. Both nicotine and a cannabinoid receptor agonist have been reported to change anxiety and both have actions on the reward process, but their effects on SNC have not been investigated. To determine: (1) whether exposure to SNC evokes an anxiogenic response; (2) whether an anxiolytic dose of nicotine has the same effects on SNC as those of chlordiazepoxide; (3) the effects of a low (anxiolytic) and a high (anxiogenic) dose of the cannabinoid receptor agonist CP 55,940 on SNC. Two groups of animals were given access to high (32%) or low (4%) sucrose solutions for 5 min per day for 10 days. On PSD 1 and 2, the shifted group had access to a devalued incentive (from 32 to 4% sucrose) and the unshifted group remained at 4% sucrose. The volumes (ml) of sucrose solution consumed were measured pre-shift and on PSD 1 and 2. In experiment 1, immediately after SNC testing on PSD 1 and 2, the rats were tested in the social interaction and elevated plus-maze tests of anxiety. In experiment 2, the effects of chlordiazepoxide (5 and 7.5 mg/kg) and nicotine (0.1 mg/kg) were examined on PSD 1 and 2. In experiment 3, the effects of CP 55,940 (5 and 40 microg/kg) were examined on PSD 1 and 2. There were no anxiogenic effects of shift in either test of anxiety on either test day. However, on PSD 1, the shifted group had significantly higher locomotor activity and spent a higher percentage of time on the open arms, perhaps reflecting search strategies. Nicotine was without significant effect on SNC on either test day. On PSD 1, chlordiazepoxide (5 mg/kg) and CP 55,940 (5 and 40 microg/kg, IP) blocked SNC. On PSD 2, both doses of chlordiazepoxide and the low, anxiolytic dose of CP 55,940 (5 microg/kg) blocked SNC, the high dose of CP 55,940 was without effect. The pattern of results allows for the separation between effects on anxiety and SNC. The block of contrast on PSD 1 was independent of changes in anxiety, since both anxiolytic and anxiogenic drug doses were effective. It is suggested that this may provide an animal model of disappointment in which the cannabinoid system plays an important role. An anxiolytic action would seem to be a necessary, but not a sufficient, action to block SNC on PSD 2.


Food preferences in human obesity: carbohydrates versus fats.
A large clinical sample of obese men and women were asked for a self-generated list of ten favorite foods. The lists were characterized by frequent instances of foods that are major nutrient sources of fat in the American diet. While obese men listed mainly protein/fat sources (meat dishes) among their favorite foods, obese women tended to list predominantly carbohydrate/fat sources (doughnuts, cookies, cake) and foods that were sweet. There was no evidence that selective preferences for a single macronutrient, carbohydrate, were a standard feature of human obesity. Rather, preferences for major nutrient sources of fat as opposed to carbohydrate may be a primary characteristic of human obesity syndromes.


Application of Berendsen barostat in dissipative particle dynamics for nonequilibrium dynamic simulation.
The Berendsen barostat from molecular dynamics simulation is applied in both standard dissipative particle dynamics (DPD) and many-body dissipative particle dynamics (MDPD) simulations. The original Berendsen barostat works well in (M)DPD simulation of a single-component system under constant pressure condition and in nonequilibrium dynamic processes. The partial Berendsen barostat is proposed for multi-component system simulation with (M)DPD. The displacement rescaling process of the Berendsen barostat is only applied on the particles outside the center region, acting as a pressure "boundary condition." The center part forms the free zone, in which the interface shape and nonequilibrium dynamic behavior between different phases can be captured properly. An immiscible bubble in the second fluid under constant pressure condition is studied, and the oscillation of the bubble radius and fluctuation of systempressure can be obtained by the current barostat. Preliminary models for bubble growing and collapsing under square pressure wave and bubble oscillation under harmonic pressure wave are also reported in the current simulation. It shows that the partial Berendsen barostat is suitable for the modeling of nonequilibrium process of single or few droplets/bubbles in multi-component systems.


Effects of poststroke losartan versus captopril treatment on myogenic and endothelial function in the cerebrovasculature of SHRsp.
We assessed the ability of poststroke captopril and losartan treatment to reverse myogenic and endothelial dysfunction in the middle cerebral arteries of Kyoto-Wistar stroke-prone spontaneously hypertensive rats (SHRsp) that developed intracerebral hemorrhagic stroke. SHRsp were sampled before and after stroke development and after up to 37 days of captopril (50 mg/kg per day) or losartan (35 mg/kg per day) treatment initiated after stroke. Pressure-dependent constriction to a 100-mm Hg pressure step, constriction to nitric oxide synthase inhibition (100 micromol/L N(omega)-nitro-l-arginine methyl ester), and endothelium-dependent vasodilation to bradykinin (1.6 micromol/L), 2-f-LIGRLO-NH(2) (1 micromol/L, a protease-activated receptor-2 agonist), and A23187 (2 micromol/L) were evaluated in middle cerebral arteries at 100 mm Hg with a pressure myograph. Middle cerebral arteries from SHRsp with stroke could not constrict to pressure or nitric oxide synthase inhibition, lacked the ability to vasodilate to bradykinin, and exhibited attenuated dilation and vasomotion in response to A23187. Vasodilation to 2-f-LIGRLO-NH(2) was unaltered. The aforementioned cerebrovascular alterations were reversed after 31 days of poststroke losartan but not of captopril treatment in the absence of an antihypertensive effect. Captopril treatment restored middle cerebral artery constriction to pressure, NOS inhibition, and bradykinin vasodilation temporarily after 7 to 18 days of treatment, after which function deteriorated to a level observed in SHRsp at stroke. Aspects of poststroke cerebrovascular dysfunction, which likely play an important role in altering and modulating cerebral blood flow autoregulation, can be reversed in SHRsp more effectively after stroke development by blocking angiotensin II type 1 receptors as opposed to lowering angiotensin II levels.


[Femoral nerve block for total knee arthroplasty].
Femoral nerve block and sciatic nerve block are used to provide intraoperative and postoperative analgesia for total knee arthroplasty. Sciatic nerve block is contraindicated in our hospital, because orthopedists want to assess peroneal nerve function after the surgery. We retrospectively assessed postoperative analgesic effect and complications of the continuous femoral nerve block for total knee arthroplasty. We included 19 cases in 17 patients scheduled to undergo total knee arthroplasty under femoral nerve block combined with general anesthesia. Ultrasound-guided femoral nerve block was performed before the surgery. The ultrasound linear probe was used to visualize the femoral nerve. A 22 gauge needle attached to a nerve stimulator, was inserted with in-plane method. Five percent glucose solution was injected through the needle to encircle the femoral nerve. Then, the 22 gauge needle was withdrawn and an 18 gauge needle was inserted with out-of-plane method. Five percent glucose solution was injected through the needle to confirm the needle tip and perineural catheter was inserted through the needle. To achieve femoral nerve block, 0.375% ropivacaine 20 ml was injected through the needle. Perineural infusion with 0.15% ropivacaine at 4 ml x hr(-1) was initiated at the end of the surgery. Intravenous patient-controlled analgesia (IV-PCA) was also conducted postoperatively. We assessed pain at rest with a verbal numeric pain rating score (0-10) including pain on moving, and nausea as well as vomiting. Patients with numeric pain scores at 3 or less were 14 out of 19. Two patients complained of severe pain. There were 4 cases suffering pain on moving. Femoral nerve separation with 5% glucose solution using in-palne method and catheter placement with out-of-plane method could be useful for perineural catheter placement. Perineural infusion of 0.15% ropivacaine at 4 ml x hr(-1) combined with IV-PCA provided a good postoperative analgesia in patients receiving total knee arthroplasty.


Protamine allergy reactions during cardiac catheterization and cardiac surgery: risk in patients taking protamine-insulin preparations.
Protamine insulin use may immunologically sensitize patients to protamine, leading to anaphylactoid reactions upon subsequent exposure to protamine sulfate during cardiac catheterization or cardiovascular surgery. The risk of such reactions in protamine insulin-dependent patients is uncertain. One catheterization study reported a 50-fold greater risk while a second showed no increased risk! To clarify the risk, the records of 7,750 cardiac catheterization procedures between 1984 and 1987 were analyzed for presence of NPH or PZI insulin use, protamine administration, and any complications or adverse reactions. Protamine was administered in 3,341/7,750 procedures (43%), including 171 in diabetics receiving NPH insulin. Adverse reactions to protamine occurred in 2/3, 170 noninsulin patients, 0.06%, and adverse reactions due to probable NPH insulin sensitization occurred in 1/171, 0.6%, of NPH diabetics, p = .034. Meta-analysis of risk showed an odds ratio of 7.96 for the NPH diabetic patients, and combining these results with the other large series in the literature (269 NPH diabetics total) showed an odds ratio of 4.19 compared to a non-NPH insulin group. Meta-analysis of the surgical literature showed the risk in surgical patients to be 2.1% in NPH patients versus 0.12% with no NPH, with an odds ratio of 15.52. The greater incidence in surgical patients may be due to protamine sensitization at prior catheterization and to the larger dose of protamine administered to surgical patients.


The Revival of an "Old" Marker: CD4/CD8 Ratio.
The effectiveness of modern antiretroviral therapies (ART) transformed HIV infection into a chronic disease characterized by a persistent condition of inflammation and immune activation. For this reason, even thought AIDS-related mortality has been reduced with an increased life expectancy, patients living with HIV infection are more likely to develop non-AIDS events despite the achievement of a complete suppression of HIV replication. Hence, the scientific community feels the need to find new biomarkers which would be suitable in clinical practice for identifying patients who require a close monitoring because of an increased risk of developing comorbidities. A renewed interest has emerged about the usefulness of CD4/CD8 ratio as a strong marker of immune activation and immune senescence. Recently, many studies have underlined that CD4/CD8 ratio might represent a good predictor of AIDS and non-AIDS events. Herein, the potential role of the CD4/CD8 ratio for the monitoring of HIV patients in different clinical settings is reviewed.


[Factors Affecting the Dynamics of Circadian Activity of Frit Flies Meromyza saltatrix (L) (Diptera: Chloropidae)].
The dynamics of circadian activity in adult frit flies of the Holarctic species Meromyza saltatrix (L) from Mongolian, Moscow, and Polish populations was studied. Synchronous peaks of activity were revealed with the periodicity multiple of three-four hours, which may depend on the level of light. The direct effect of temperature and humidity on the activity of flies outside the optimal values of these factors was found. It was detected that the peak of adult emergence falls on the beginning of a general increase in the abundance of flies, which indicates constant rejuvenation of the population. The sex ratio is close to 1, but the emergence of males and females is in antiphase. The synchronization of peaks of circadian activity in the populations from different regions confirms the presence of a circadian rhythm of activity. The rhythm synchronizing the reproductive activity of adults was found to be modified by the photoperiod under the optimum conditions of temperature and humidity.


Decreased metabotropic glutamate receptor type 1 availability in a patient with spinocerebellar ataxia type 6: A (11)C-ITMM PET study.
Imaging of metabotropic glutamate receptor type 1 (mGluR1), localized exclusively in the cerebellar Purkinje cells and related to cerebellar function, has recently become possible using positron emission tomography (PET). We report the initial mGluR1 imaging in a 74-year-old woman with spinocerebellar ataxia type 6 (SCA6). The patient and 9 age-matched healthy controls underwent PET scanning with a mGluR1 radiotracer, N-[4-[6-(isopropylamino)pyrimidin-4-yl]-1,3-thiazol-2-yl] -4-(11)C-methoxy-N-methylbenzamide. Volumes-of-interest were placed on the anterior and posterior lobes, vermis, and flocculus. Binding potential (BPND) was calculated to estimate mGluR1 availability using the simplified reference tissue model. A partial volume correction was applied to the BPND values. Additionally, the volume of the whole cerebellum was measured using MRI. The corrected BPND values of the cerebellar subregions and the volume of the whole cerebellum in the patient were 51.0% to 68.3% and 72.6%, respectively, of the controls. Thus, the magnitude of reduced BPND values was relatively larger than the magnitude of cerebellar atrophy in the patient. These findings suggest that the measurement of mGluR1 availability is more sensitive than morphological measurements by MRI to detect reduced cerebellar function. Thus, imaging of mGluR1, probably reflecting the number and distribution of Purkinje cells, can be a specific and sensitive marker for estimation of cerebellar function.


Caveolae preservation in the characterization of human neuromuscular disease.
We have examined freeze-fracture replicas and conventional thin-section images of rat myofibers prepared by perfusion and by conventional immersion fixation protocols, and myofibers of normal and dystrophic human myofibers prepared by similar immersion fixation methods. In both rat and human myofibers, the size and distribution of caveolae was found to differ substantially according to the method of glutaraldehyde exposure, the depth of the myofiber from the surface exposed to the fixative, and if surgically bisected, the distance from the cut end of the myofiber. Conventional immersion fixation resulted in unavoidable but predictable alterations in sarcolemmal caveolae. These reproducible artifacts of fixation technique substantially complicate the use of caveolae as reliable markers for the characterization of human neuromuscular disease.


[Daily practice using the guidelines for prevention and treatment of osteoporosis. Semi-quantitative method in the assessment of vertebral fracture].
Assessment of vertebral fracture based on a thoraco-lumbar spine radiographs is a good tool to evaluate the risk of incident fracture. Presence of vertebral fracture indicates 4 to 7 fold increase of risk of future fracture. However, vertebral fracture are often underdiagnosed. Quantitative (QM) and semi-quantitative (SQ) methods are designed for the assessment of prevalent and incident fracture. The advantages of SQ method are more convenient and reproducible as well as more excellent in the assessment of the risk of future fracture compared to QM. It should be recognized that untrained SQ reader produce a high false negative of grade 1 (mild fracture).


The impact of HLA-DR antigen matching on the survival of cadaveric renal allografts. A prospective one-center analysis.
The impact of HLA-DR antigen matching on the survival of cadaveric renal allografts was assessed in 158 consecutive transplants performed in our unit since early 1978. In 41 donor-recipient pairs with two shared HLA-DR antigens, the actuarial graft survival rate at 6 months was 73% as compared with 51% in 76 transplants with one HLA-DR antigen shared and 32% in 41 transplants with zero shared HLA-DR antigens. This finding is highly significant (P for heterogeneity [PH] = 0.0005 and P for trend, [PT] = 0.0001). Our data clearly indicate that HLA-DR antigen sharing is more beneficial than merely avoiding HLA-DR incompatibility. But, the frequent antigen HLA-DRw6 was not taken into account in this study due to difficulties in its identification. We found no evidence that the observed beneficial effect of HLA-DR matching could be explained by interaction of other prognostic factors, such as sex, age, previous transplantation, diabetes mellitus or pretransplant blood transfusion. Patients who did not receive blood transfusion prior to transplantation had a significantly lower graft survival rate than those who did (13% vs. 56% at 6 months). HLA-DR matching was found to have a powerful effect on graft survival even among pretransplant blood transfused recipients (PH = 0.002, PT = 0.0006). We conclude that selection of recipients for transplantation should attempt to achieve HLA-DR identical combinations.


Longitudinal Relationships Between Reflective Functioning, Empathy, and Externalizing Behaviors During Adolescence and Young Adulthood.
Reflective functioning (RF) refers to the understanding of one's own and others' behaviors in terms of mental states, whereas empathy entails the abilities to understand (cognitive empathy) and to share (affective empathy) the emotions of others. Low RF and low empathy have been previously related to externalizing behaviors, such as aggression and rule breaking. However, few longitudinal studies have simultaneously examined the relationships between these variables during adolescence. The aim of the present study is to investigate the longitudinal effects of both RF and empathy on potential changes in externalizing behaviors over time, in a group of 103 adolescents and young adults from the general population assessed repeatedly up to four times. We conducted multilevel analysis in order to examine the effects of RF and empathy on the initial levels and the trajectories of externalizing behaviors over time, while accounting for other variables previously associated with externalizing behaviors, such as age, gender, internalizing problems, and cognitive abilities. The results suggest that the ability to reflect on behaviors in terms of mental states predicted a sharper decrease in externalizing behaviors over time. Moreover, externalizing behaviors at the first assessment were associated with RF impairments and low affective empathy. Age, gender, cognitive abilities, and cognitive empathy were not associated with externalizing behaviors. We discuss how our results, based on a typically developing population, might inform primary or indicated prevention strategies for externalizing behaviors by focusing on socio-cognitive processes such as RF and affective empathy.


[The characteristics of postloading bronchospasm in bronchial asthma patients].
A study of 73 patients suffering of asthma bronchiale with postloading bronchospasm and 20 without bronchospasm indicates that most pronounced bronchoconstriction after physical loads appears on the 15-20 minute in atopic asthma and asthma of physical exertion and by the 5-10 min. in infectious-allergic form of the disease. It is recommended to use MEF75, MEF50 and MEF25 as a diagnostic criterion of postloading bronchospasm.


Behavior of amino acids when volatilized in the presence of silica gel and pulverized basaltic lava.
To evaluate the types of amino acid thermal transformations caused by silicate materials, we studied the volatilization products of Aib, L-Ala, L-Val and L-Leu under temperatures of up to 270 degrees C in the presence of silica gel as a model catalyst and pulverized basaltic lava samples. It was found that silica gel catalyzes nearly quantitative condensation of amino acids, where piperazinediones are the major products, whereas lava samples have much lower catalytic efficiency. In addition bicyclic and tricyclic amidines and several products of their subsequent thermal decomposition have been identified using the coupled technique of GC-FTIR-MS and HPLC-PB-MS, with auxiliary computer simulation of IR spectra and NMR spectroscopy. The decomposition is due to dehydrogenation, elimination of the alkyl substituents and dehydration as well as cleavage of the bicyclic ring system. The imidazole ring appears to be more resistant to thermal decomposition as compared to the pyperazine moiety, giving rise to the formation of different substituted imidazolones. The amidines were found to hydrolyze under treatment with concentrated HCl, releasing the starting amino acids and thus behaving as amino acid anhydrides. The thermal transformations cause significant racemization of amino acid residues. Based on our observations, the formation of amidine-type products is suggested to be rather common in the high-temperature experiments on amino acid condensation.


Three dimensional water quality modeling of a shallow subtropical estuary.
Knowledge of estuarine hydrodynamics and water quality comes mostly from studies of large estuarine systems. The processes affecting algae, nutrients, and dissolved oxygen (DO) in small and shallow subtropical estuaries are relatively less studied. This paper documents the development, calibration, and verification of a three dimensional (3D) water quality model for the St. Lucie Estuary (SLE), a small and shallow estuary located on the east coast of south Florida. The water quality model is calibrated and verified using two years of measured data. Statistical analyses indicate that the model is capable of reproducing key water quality characteristics of the estuary within an acceptable range of accuracy. The calibrated model is further applied to study hydrodynamic and eutrophication processes in the estuary. Modeling results reveal that high algae concentrations in the estuary are likely caused by excessive nutrient and algae supplies in freshwater inflows. While algal blooms may lead to reduced DO concentrations near the bottom of the waterbody, this study indicates that stratification and circulation induced by freshwater inflows may also contribute significantly to bottom water hypoxia in the estuary. It is also found that high freshwater inflows from one of the tributaries can change the circulation pattern and nutrient loading, thereby impacting water quality conditions of the entire estuary. Restoration plans for the SLE ecosystem need to consider both a reduction of nutrient loading and regulation of the freshwater discharge pattern.


Morphological plasticity in the chick ventral lateral geniculate nucleus: temporal parameters.
The retinogeniculate projection in the chick undergoes apparent augmentation following lesions in the optic tectum. Using autoradiographic tracing techniques we determined that the alteration of the retinal projection required a minimum of 4 days to be detected if tectal lesions were made at hatching and could be produced by lesions placed up to 1.5 years posthatch.


[Doctor-Patient communication].
The Doctor-Patient relationship is of fundamental importance in the field of care and continues to undergo profound transformations. In ancient times the doctor was "authoritarian": the patient was considered a passive element, incapable of understanding, knowing and acting for his own health. The doctor then became a "paternalistic" figure and it is only recently that the patient has become a responsible subject who interacts with the doctor in the definition and evaluation of every health intervention. The patient needs to be actively involved and informed, participating in the therapeutic process and in monitoring the positive and negative effects of the treatment. The doctor needs to communicate in a way that is simple and appropriate in order to avoid misunderstandings, which are very common as patients and doctors can often attribute different meanings to certain words. Doctors need to be aware that such a possibility exists and verify that the patient has understood the real meaning of his words. As good communication between doctors and patients is a strategic factor in care, communication courses should be included in any Medical Degree course.


Integration of new CAPS and dCAPS-RGA markers into a composite chickpea genetic map and their association with disease resistance.
A composite linkage map was constructed based on two interspecific recombinant inbred line populations derived from crosses between Cicer arietinum (ILC72 and ICCL81001) and Cicer reticulatum (Cr5-10 or Cr5-9). These mapping populations segregate for resistance to ascochyta blight (caused by Ascochyta rabiei), fusarium wilt (caused by Fusarium oxysporum f. sp. ciceris) and rust (caused by Uromyces ciceris-arietini). The presence of single nucleotide polymorphisms in ten resistance gene analogs (RGAs) previously isolated and characterized was exploited. Six out of the ten RGAs were novel sequences. In addition, classes RGA05, RGA06, RGA07, RGA08, RGA09 and RGA10 were considerate putatively functional since they matched with several legume expressed sequences tags (ESTs) obtained under infection conditions. Seven RGA PCR-based markers (5 CAPS and 2 dCAPS) were developed and successfully genotyped in the two progenies. Six of them have been mapped in different linkage groups where major quantitative trait loci conferring resistance to ascochyta blight and fusarium wilt have been reported. Genomic locations of RGAs were compared with those of known Cicer R-genes and previously mapped RGAs. Association was detected between RGA05 and genes controlling resistance to fusarium wilt caused by races 0 and 5.


Advances in signalling by extracellular nucleotides. the role and transduction mechanisms of P2Y receptors.
Nucleotides are ubiquitous intercellular messengers whose actions are mediated by specific receptors. Since the first clonings in 1993, it is known that nucleotide receptors belong to two families: the ionotropic P2X receptors and the metabotropic P2Y receptors. Five human P2Y receptor subtypes have been cloned so far and a sixth one must still be isolated. In this review we will show that they differ by their preference for adenine versus uracil nucleotides and triphospho versus diphospho nucleotides, as well as by their transduction mechanisms and cell expression.


Transglutaminase-induced crosslinking of gelatin-calcium carbonate composite films.
The effects of transglutaminase (TGase) on the rheological profiles and interactions of gelatin-calcium carbonate solutions were studied. In addition, mechanical properties, water vapour permeability and microstructures of gelatin-calcium carbonate films were also investigated and compared. Fluorescence data suggested that the interaction of TGase and gelation-calcium carbonate belonged to a static quenching mechanism, and merely one binding site between TGase and gelatin-calcium carbonate was identified. Moreover, differential scanning calorimetry (DSC), the mechanical properties and the water vapour permeability studies revealed that TGase favoured the strong intramolecular polymerisation of the peptides in gelatin. The microstructures of the surfaces and cross sections in gelatin-calcium carbonate films were shown by scanning electron microscope (SEM) micrographs. The results of the fourier transform infrared spectroscopy (FTIR) indicated that TGase caused conformational changes in the proteins films. Therefore, TGase successfully facilitated the formation of gelatin-calcium carbonate composite films.


[Identification of idiotrophs for 3'-deoxykanamycin B synthesis in Streptomyces tenebrarius (Higgens a. Kastner)].
Mutants of Streptomyces tenebrarius with the blocked synthesis of 3'-deoxykanamycin B were obtained by treating the producer with NTG and chloramphenicol, or after gamma-irradiation. These mutants (idiotrophs) were distributed into three groups by means of the cosynthesis experiments on agar plates: convertors, secretors and "neutral" strains. Five idiotrophs represented five complementation groups for biosynthesis of the antibiotic. Three of these were defective in 2-deoxystreptamine synthesis, the fourth was defined as neamine-negative, and the fifth was probably blocked in regulation of enzymes responsible for conversion of neamine or paromamine into kanamycins. Localization of mutations has been shown on the scheme of kanamycins' biosynthesis.


Compulsive exercise as a mediator between clinical perfectionism and eating pathology.
The aim of this study was to examine whether compulsive exercise mediates the relationship between clinical perfectionism and eating pathology, based on the cognitive behavioral model of compulsive exercise. Participants were 368 adults who participated regularly in sport/exercise and completed online measures of perfectionism, compulsive exercise and eating disorders. In support of the well-established link between perfectionism and eating disorders, clinical perfectionism predicted eating pathology both directly and indirectly mediated by compulsive exercise. In addition, there were also direct effects of clinical perfectionism on the avoidance/rule-driven behavior, weight control, and mood improvement subscales of the Compulsive Exercise Test (CET). There was a direct effect of the CET weight control subscale on eating pathology and a negative direct effect of the CET subscale mood improvement on eating pathology. Findings lend support to the cognitive behavioral model of compulsive exercise in which clinical perfectionism is conceptualized as related to eating disorders directly and indirectly through the mediation of compulsive exercise. Compulsive exercise was also found to have a direct effect on eating disorders. Compulsive exercise may be a symptom of eating pathology, rather than an antecedent, however causal inferences could not be established given the correlational design. Longitudinal research using cross-lagged panel designs to examine a bidirectional relationship between compulsive exercise and eating disorders is needed.


Continuous wave and Q-switched laser output of laser-diode-end-pumped disordered Nd:LGS laser.
Laser-diode (LD)-pumped high-power cw and passively Q-switched laser performance has been demonstrated with a disordered Nd-doped La(3)Ga(5)SiO(14) (Nd:LGS) crystal for the first time to our knowledge. A cw output power of 2.25 W was obtained under an absorbed pump power of 7.84 W with a slope efficiency of 30.2%. Using a Cr:YAG as a saturable absorber, the passively Q-switched laser was achieved. The maximum pulse energy, shortest pulse width, and highest peak power were measured to be 117.5 muJ, 23.4 ns, and 5.02 kW, respectively.


The carboxyl terminus of zona occludens-3 binds and recruits a mammalian homologue of discs lost to tight junctions.
Mammalian homologues of the Drosophila polarity proteins Stardust, Discs Lost, and Crumbs have been identified as Pals1, Pals1-associated tight junction protein (PATJ), and human Crumbs homologue 1 (CRB1), respectively. We have previously demonstrated that PATJ, Pals1, and CRB1 can form a tripartite tight junction complex in epithelial cells and that PATJ recruits Pals1 to tight junctions. Here, we observed that the Pals1/PATJ interaction was not crucial for the ultimate targeting of PATJ itself to tight junctions. This prompted us to examine if any of the 10 post-synaptic density-95/Discs Large/zona occludens-1 (PDZ) domains of PATJ could bind to the carboxyl termini of known tight junction constituents. We found that the 6th and 8th PDZ domains of PATJ can interact with the carboxyl termini of zona occludens-3 (ZO-3) and claudin 1, respectively. PATJ missing the 6th PDZ domain was found to mislocalize away from cell contacts. Surprisingly, deleting the 8th PDZ domain had little effect on PATJ localization. Finally, reciprocal co-immunoprecipitation experiments revealed that full-length ZO-3 can associate with PATJ. Hence, the PATJ/ZO-3 interaction is likely important for recruiting PATJ and its associated proteins to tight junctions.


Creating the basis for a breast health program for female survivors of Hodgkin disease using a participatory research approach.
To learn from female survivors of Hodgkin disease about their perceptions of their current health status and future health risks, self-care practices to prevent or diminish health risks, and what kind of breast health program could benefit them, including the most effective methods and optimal times for learning about breast health. Participatory research using focus groups. Urban pediatric cancer center. 1 African American and 19 Caucasian female survivors of Hodgkin disease aged 16-26 years, diagnosed at least two years before the start of the study, and treated with mantle radiation therapy. Participants were recruited during visits to an outpatient clinic. Six open-ended questions were asked during three separate focus group sessions. Transcribed data were evaluated by content analysis techniques and analyzed to identify common themes. Current health status and perceived health risks, current health practices, and effective methods and timing for breast health teaching. Survivors reported feeling damaged by their cancer and its treatment and perceived that they were at risk for breast cancer. Self-care and risky behaviors also were reported. Internal influences (e.g., fear) and external influences (e.g., family) motivated survivors to participate in health promotion activities. Effective methods identified for learning about breast health included having access to other survivors, being respected as an adult, and having one-on-one staff teaching and peer support. The preferred timing of teaching varied, but survivors generally supported a gradual provision of information. A positive listening environment is important for developing a breast health program for survivors. An essential first step is to create an opportunity for survivors to tell about their experiences with cancer, including its impact on their lives. Information regarding breast health must be provided in multiple formats during and after treatment if good practices are to be undertaken. The provision of adequate information during and after therapy as well as peer counseling in a positive listening environment are important in helping survivors participate in health promotion activities.


Perceptions and effects of antenatal education.
To assess the quality and effect of antenatal health education on health-related issues during pregnancy, and the benefits to women during the peri-partum period. This is a descriptive study of 300 women attending the Postnatal Clinics and Women Health Clinics in 2 Family Medicine centers at King AbdulAziz Medical City, National Guard Health Affairs, Riyadh, Saudi Arabia. The study was carried out between November 2009 and February 2010. A predesigned questionnaire was used. Three hundred women completed the questionnaire. Most women made their first antenatal visit during the fourth month of pregnancy, and most had 3-6 antenatal care (ANC) visits. Highly educated mothers had significantly more ANC visits and attended the first visit earlier. Nearly 80% of subjects were satisfied with the health education provided during ANC visits. Multiparous women were found to receive less health education than primiparous women. Most reported receiving education on breast feeding (83%), signs of labor (75.3%), and baby care (74.7%). However, most of the subjects did not receive all of the important information that should be given during ANC. There was a negative linear relationship between health education score of women and their age. This study quantified the inadequate level of health education received by pregnant women during ANC.


Sex-specific cortisol levels in bipolar disorder and schizophrenia during mental challenge--relationship to clinical characteristics and medication.
Our objective was to examine the cortisol release during a mental challenge in severe mental disorders versus healthy controls (HC), analyzing effects of sex, clinical characteristics and medication, and comparing Bipolar Disorder (BD) to Schizophrenia (SCZ). Patients with BD and SCZ (n=151) were recruited from a catchment area. HC (n=98) were randomly selected from the same area. Salivary samples were collected before and after a mental challenge and cortisol levels determined. During the challenge there was an interaction between group and sex (P = 0.015) with male patients having a blunted cortisol release compared to male HC (P = 0.037). Cortisol change did not differ significantly between BD and SCZ. In all patients, the cortisol change correlated with number of psychotic episodes (r = -0.23, P = 0.025), and in females patients, with number of depressive episodes (r = -0.33, P = 0.015). Patients using antidepressants had a greater cortisol release during challenge than those not using antidepressants (P = 0.043). Male patients with severe mental disorders seem to have a uniform abnormal cortisol release during mental challenges which associates with clinical course, and with beneficial effects of antidepressants.


Effect of administration route on the renal safety of contrast agents: a meta-analysis of randomized controlled trials.
The objective of this study was to examine the association between administration route and relative renal safety of contrast agents. We searched all published articles indexed in Embase, Medline and the Cochrane Central Register of Controlled Trials, from January 1980 to November 2010, to identify relevant studies. Of the 1,047 initially identified studies, 11 randomized controlled trials (RCTs) including 2,210 patients with intra-arterial route and 7 RCTs including 919 patients with intravenous route were finally analyzed. With regard to intra-arterial route, our meta-analysis showed that iodixanol significantly decreased the risk of contrast-induced acute kidney injury (CI-AKI) when compared with a pool of low-osmolar contrast media (LOCM; risk ratio [RR] = 0.68; 95% confidence interval [95% CI], 0.50-0.92; Z=2.47; p=0.01), with no significant heterogeneity between individual studies (p=0.14, I2=32.4%). However, iodixanol was not associated with a reduction in CI-AKI compared with the LOCM pooled together (RR=0.75; 95% CI, 0.44-1.26; Z=1.10; p=0.27) with intravenous application, again with no significant heterogeneity between individual studies (p=0.40, I2=3.6%). Our meta-analysis suggests that administration route may affect the renal safety of contrast agents. Specifically, iodixanol may be a better choice for patients in the interventional cardiology setting.


Reaction pathways for the photodegradation of the organophosphorus cyanophos in aqueous solutions.
Photodegradation in aqueous solutions is an important pathway for many agrochemicals such as pesticides. In the present work, the photochemical transformation of cyanophos (CYA) was investigated in aqueous solutions using UV light within the 254-313 nm range as well as solar light. The study was performed in order to have a deep insight into the mechanistic pathways for the photochemical disappearance of CYA. Upon UV irradiation of an aerated solution of CYA, the degradation quantum yield was found equal to 1.8 x 10(-2). It is independent of the excitation wavelength but varies with oxygen concentration. It increased by a factor of 2 from oxygen-saturated to oxygen-free solution. Photosensitized experiments were performed using acrylamide and hydroquinone as energy acceptor and energy donor substrates, respectively. They show that both singlet and triplet excited states were involved in the photochemical behavior of CYA. The laser flash photolysis experiments clearly showed the involvement of the triplet excited state which was efficiently quenched by molecular oxygen and acrylamide with the rate constants 1.97 x 10(9) and 2.71 x 10(9) mol(-1) L s(-1), respectively. The photoproducts structures were proposed according to the mass spectral data using the LC/MS technique. The analytical study shows that various processes such as hydrolysis, homolytic bond dissociations and Photo-Fries process occur.


Fluorometric determination of DNA in agarose gels: usefulness for measurement of double-strand breaks in nonlabeled cells by pulsed-field gel electrophoresis.
Quantitative measurement of DNA in agarose gels, particularly as needed for measurement of double-strand breaks induced by agents such as radiation, usually involves the use of radioactively labeled DNA. Thus its usefulness is usually limited to growing cells which incorporate radiolabeled thymidine into DNA. To circumvent this problem, we have developed a fluorometric technique for quantitative estimation of DNA in the presence of large amounts of agarose. Gel slices are solubilized with concentrated sodium perchlorate and DNA is selectively precipitated with cadmium chloride. The amount of DNA can then be estimated with 3,5-diaminobenzoic acid. Determination of DNA is linear in the range 10 ng to 1 microgram or more. We describe the application of this technique to the measurement of 60Co gamma-ray-induced double-strand breaks by pulsed-field gel electrophoresis. Our results are essentially identical to those obtained using radiolabeled DNA.


Oxaliplatin-induced pancreatitis: a case series.
Drug-induced pancreatitis is less common compared to other causes of acute pancreatitis; the incidence ranges from between 0.1% to 2% of acute pancreatitis cases. Among alkylating agents, oxaliplatin has not been reported to be associated with acute pancreatitis. This case study presents a series of six cases of acute pancreatitis presumably related to exposure to oxaliplatin which had different gastrointestinal malignancies and were being treated with oxaliplatin in combination with other chemotherapeutic drugs. All other related causes of acute pancreatitis were excluded before implicating oxaliplatin as a possible cause. In all cases, oxaliplatin was stopped and patients had resolution of their signs and symptoms, along with a decrease in serum amylase and lipase levels. Knowledge regarding acute pancreatitis related to oxaliplatin is of paramount importance in order to diagnose cases early and institute effective treatment in patients who are undergoing chemotherapy with this drug.


The peroxisome proliferator-activated receptor (PPAR) β/δ agonist GW501516 inhibits IL-6-induced signal transducer and activator of transcription 3 (STAT3) activation and insulin resistance in human liver cells.
IL-6 induces insulin resistance by activating signal transducer and activator of transcription 3 (STAT3) and upregulating the transcription of its target gene SOCS3. Here we examined whether the peroxisome proliferator-activated receptor (PPAR)β/δ agonist GW501516 prevented activation of the IL-6-STAT3-suppressor of cytokine signalling 3 (SOCS3) pathway and insulin resistance in human hepatic HepG2 cells. Studies were conducted with human HepG2 cells and livers from mice null for Pparβ/δ (also known as Ppard) and wild-type mice. GW501516 prevented IL-6-dependent reduction in insulin-stimulated v-akt murine thymoma viral oncogene homologue 1 (AKT) phosphorylation and in IRS-1 and IRS-2 protein levels. In addition, treatment with this drug abolished IL-6-induced STAT3 phosphorylation of Tyr⁷⁰⁵ and Ser⁷²⁷ and prevented the increase in SOCS3 caused by this cytokine. Moreover, GW501516 prevented IL-6-dependent induction of extracellular-related kinase 1/2 (ERK1/2), a serine-threonine protein kinase involved in serine STAT3 phosphorylation; the livers of Pparβ/δ-null mice showed increased Tyr⁷⁰⁵- and Ser⁷²⁷-STAT3 as well as phospho-ERK1/2 levels. Furthermore, drug treatment prevented the IL-6-dependent reduction in phosphorylated AMP-activated protein kinase (AMPK), a kinase reported to inhibit STAT3 phosphorylation on Tyr⁷⁰⁵. In agreement with the recovery in phospho-AMPK levels observed following GW501516 treatment, this drug increased the AMP/ATP ratio and decreased the ATP/ADP ratio. Overall, our findings show that the PPARβ/δ activator GW501516 prevents IL-6-induced STAT3 activation by inhibiting ERK1/2 phosphorylation and preventing the reduction in phospho-AMPK levels. These effects of GW501516 may contribute to the prevention of cytokine-induced insulin resistance in hepatic cells.


Combined 3D-QSAR modeling and molecular docking study on quinoline derivatives as inhibitors of P-selectin.
P-selectin is a promising target for developing novel atherosclerosis drugs. To understand the structure-activity correlation of quinolines-based P-selectin inhibitors, we have carried out a combined molecular docking and three-dimensional quantitative structure-activity relationship (3D-QSAR) modeling study. The study has resulted in two types of satisfactory 3D-QSAR models, including the CoMFA model (r(2), 0.863; q(2), 0.589) and CoMSIA model (r(2), 0.866; q(2), 0.636), to predict the biological activity of new compounds. The detailed microscopic structures of P-selectin binding with inhibitors have been studied by molecular docking. We have also developed docking based 3D-QSAR models (CoMFA with r(2), 0.934; q(2), 0.591; CoMSIA with r(2), 0.896; q(2), 0.573). The contour maps obtained from the 3D-QSAR models in combination with the docked binding structures help to better interpret the structure-activity relationship. All of the structural insights obtained from both the 3D-QSAR contour maps and molecular docking are consistent with the available experimental activity data. The satisfactory results strongly suggest that the developed 3D-QSAR models and the obtained P-selectin-inhibitor binding structures are reasonable for the prediction of the activity of new inhibitors and in future drug design.


Unilateral once daily milking locally induces differential gene expression in both mammary tissue and milk epithelial cells revealing mammary remodeling.
Once daily milking reduces milk yield, but the underlying mechanisms are not yet fully understood. Local regulation due to milk stasis in the tissue may contribute to this effect, but such mechanisms have not yet been fully described. To challenge this hypothesis, one udder half of six Holstein dairy cows was milked once a day (ODM), and the other twice a day (TDM). On the 8th day of unilateral ODM, mammary epithelial cells (MEC) were purified from the milk using immunomagnetic separation. Mammary biopsies were harvested from both udder halves. The differences in transcript profiles between biopsies from ODM and TDM udder halves were analyzed by a 22k bovine oligonucleotide array, revealing 490 transcripts that were differentially expressed. The principal category of upregulated transcripts concerned mechanisms involved in cell proliferation and death. We further confirmed remodeling of the mammary tissue by immunohistochemistry, which showed less cell proliferation and more apoptosis in ODM udder halves. Gene expression analyzed by RT-qPCR in MEC purified from milk and mammary biopsies showed a common downregulation of six transcripts (ABCG2, FABP3, NUCB2, RNASE1 and 5, and SLC34A2) but also some discrepancies. First, none of the upregulated transcripts in biopsies varied in milk-purified MEC. Second, only milk-purified MEC showed significant LALBA downregulation, which suggests therefore that they correspond to a mammary epithelial cell subpopulation. Our results, obtained after unilateral milking, suggest that cell remodeling during ODM is due to a local effect, which may be triggered by milk accumulation.


Body composition of healthy 7-and 8-year-old children and a comparison with the 'reference child'.
There are few longitudinal data on body composition in healthy children. This has prompted a reliance on notional standards such as the 'reference child', to validate new methods of determining body composition and comparing cross-sectional height, weight and fatness data. These were twofold-to provide normative longitudinal data on changes in body composition in healthy pre-pubertal children, and to compare measures of growth and body composition with the appropriate age-specific reference child. A sample of healthy Scottish children aged 7-8y (n = 257) was recruited during 1991/1992. Data on height, weight, skinfold thickness and resistance from bioelectrical impedance analysis were collected twice, 12 months apart. Percentage body fat was estimated from both skinfolds and bioelectrical impedance. Fat and fat-free mass, but not body mass index, differed between boys and girls. All measurements increased significantly over the 12 month period except percentage body fat from skinfolds in boys. The reference child comparison revealed that our sample was taller, heavier and fatter and gained weight and fat mass at a greater rate than the Fomon standards. Data from the children in this study suggest that the reference child has a body composition which is now out of date. This may have important implications for body composition methodology. New references for height and weight may be required, but an upgrading of the body fat reference may conflict with public health aims to reduce obesity.


Silver oxide ointment wound dressing in venous ulcerations: home, self-management.
Venous ulcers are a common, chronic medical and social problem. These ulcers are difficult to heal in most patients with sustained venous hypertension. This pilot product evaluation registry study has evaluated the efficacy and safety of an antimicrobial silver oxide wound dressing ointment ("Silver Oxide Ointment") as part of the treatment of 'difficult' venous ulcerations. The study was conducted measuring the variations in the area of the ulceration and microcirculatory parameters. After four weeks, treatment with the Silver Oxide Ointment proved more effective than the 'best management' used in controls. Transcutaneous PO2 was increased (improved); Laser Doppler skin flux and transcutaneous PCO2 were improved (decreased). Also in the silver oxide group a significantly higher number of venous ulcers were completely healed at four weeks. The silver oxide ointment improved both the microcirculation and the healing rate of their ulcers. No significant tolerability problems were observed. In difficult venous ulcerations, local treatment with this Silver Oxide Ointment, as one component of the total wound dressing, improved microcirculation measurements and healing rate. Most of the treatments were done at home by the patients or by their tutors. This study indicates the important role of this type of treatment and indicates the need to plan larger and more prolonged studies.


Synthesis, structural analysis, and magnetic properties of ethylmalonate-manganese(II) complexes.
Five manganese(II) complexes of formulas [Mn(2)(Etmal)(2)(H(2)O)(2)(L)](n) (1-4) and {[Mn(Etmal)(2)(H(2)O)][Mn(H(2)O)(4)]}(n) (5) with H(2)Etmal = ethylmalonic acid (1-5) and L = 1,2-bis(4-pyridyl)ethane (bpa) (1), 4,4'-azobispyridine (azpy) (2), 4,4'-bipyridyl (4,4'-bpy) (3), and 1,2-bis(4-pyridyl)ethylene (bpe) (4) were synthesized and structurally characterized by single crystal X-ray diffraction. Their thermal behavior and variable-temperature magnetic properties were also investigated. The structure of the compounds 1-4 consists of corrugated layers of aquamanganese(II) units with intralayer carboxylate-ethylmalonate bridges in the anti-syn (equatorial-equatorial) coordination mode which are linked through bis-monodentate bpa (1), azpy (2), 4,4'-bpy (3), and bpe (4) ligands to build up a three-dimensional (3D) framework. The structure of compound 5 is made up by zigzag chains of manganese(II) ions with a regular alternation of [Mn(H(2)O)(4)](2+) and chiral (either Δ or λ enantiomeric forms) [Mn(Etmal)(2)(H(2)O)](2-) units within each chain. In contrast to the bidentate/bis-monodentate coordination mode of the Etmal ligand in 1-4, it adopts the bidentate/monodentate coordination mode in 5 with the bridging carboxylate-ethylmalonate also exhibiting the anti-syn conformation but connecting one equatorial and an axial position from adjacent metal centers. The manganese-manganese separation through the carboxylate-ethylmalonate bridge in 1-5 vary in the range 5.3167(4)-5.5336(7) Å. These values are much shorter than those across the extended bis-monodentate N-donors in 1-4 with longest/shortest values of 11.682(3) (3)/13.9745(9) Å (4). Compounds 1-5 exhibit an overall antiferromagnetic behavior, where the exchange pathway is provided by the carboxylate-ethylmalonate bridge. Monte Carlo simulations based on the classical spin approach (1-5) were used to successfully reproduce the magnetic data of 1-5.


Genetic characterization and isolation of the Saccharomyces cerevisiae gene coding for uridine monophosphokinase.
We selected a 5-fluorouracil-resistant, thermosensitive mutant of the uridine monophosphokinase step in Saccharomyces cerevisiae. The mutant displays very weak thermolabile uridine monophosphokinase activity and wild-type uridine diphosphokinase activity. Growth of the mutant at the non-permissive temperature causes immediate reduction of pyrimidine triphosphate pools to 10% of the wild-type level as well as significantly lowering total RNA and protein synthesis. These conditions also provoke derepression of the first gene of the pathway, URA2, at both the levels of enzymatic activity and transcription. The mutation segregates independently of all known genes of the pyrimidine biosynthetic pathway. The corresponding gene has been isolated on a 4.8 kb fragment by complementation of the mutant phenotype. The new gene, named URA6, codes for a 2.2 kb polyadenylated messenger RNA, exists in a single copy per haploid genome, and was mapped to the centromere of chromosome XI.


Development and psychometric evaluation of a safety climate scale for vineyards.
This study aimed to develop a questionnaire-based tool measuring the safety climate in vineyards and to assess its psychometric properties. A literature search was conducted to identify the dimensions and items that constitute the safety culture construct in various occupational sectors and to draft a conceptual framework. Content validity appraisal was performed by 16 farm managers or pesticide operators. The resulting preliminary conceptual framework consisted of 9 dimensions and 42 questions. Then, a telephone survey was conducted in the French Aquitaine (Bordeaux) region with 312 vineyard workers. Item-total correlation tests, Cronbach's alpha analysis and a principal component analysis were performed to confirm the unidimensionality of the scale under construction. Structural equation modeling (SEM) techniques were used to verify the model hypothesized from the exploratory analyses and to determine how well it fits the data. Exploratory analyses resulted initially in a 9-dimension, 20-item safety climate questionnaire. Internal consistency proved good with a Cronbach's alpha equal to 0.81. The SEM approach suggested two dimension groupings for a better fit of the data (7 dimensions operationalized through the same 20 items). Internal model parameters showed that the more influential dimensions of safety climate were Management commitment, Communication and feedback, Rules and practices, and Knowledge (all standardized path coefficients ≥ 0.7). Owing to its good psychometric properties, we hope this score will help in drawing up relevant interventions aimed at improving safety culture, raising pesticide risk awareness, and hopefully inducing more sustainable practices in the medium-term future.


Immunocytochemical localization of calmodulin in PC12 cells and its possible interaction with histones.
The subcellular localization of calmodulin, a multi-functional calcium-binding regulatory protein, was examined immunocytochemically in undifferentiated PC12 rat pheochromocytoma cells and cells differentiated with nerve growth factor (NGF) and dibutyryl cyclic AMP. In undifferentiated PC12 cells, diffuse immunostaining for calmodulin was observed in the cytoplasm, and weak, patch-like staining was found in the nucleus. In differentiated cells, intense immunostaining for calmodulin was observed in the cytoplasm, while nuclear immunostaining was still evident. Immunoreactivity for calmodulin was also observed along newly-formed neuritic processes, with strong staining in varicosity-like structures and growth cones. Using double-label immunochemistry, the relative intensity of immunostaining for calmodulin among the nuclei was found to correlate with the relative intensity of immunostaining for histones in the same nuclei. A comparison of a profile of 125I-calmodulin binding in the nuclear fraction from PC12 cells to that of immunoblotting for histones in the same fraction indicated that some of the histones are calmodulin-binding proteins in PC12 cells. These results show that the level and subcellular distribution of calmodulin are altered during the course of nerve cell differentiation and suggest the possibility that histones may function as major nuclear binding proteins for calmodulin.


Psychometric properties of 12-item self-administered World Health Organization disability assessment schedule 2.0 (WHODAS 2.0) among general population and people with non-acute physical causes of disability - systematic review.
Objective: WHODAS 2.0 is a unified scale to measuring disability across diseases, countries, and cultures. The objective was to explore the available evidence on the psychometric properties of 12-item self-administered WHODAS 2.0 among a general population and people with non-acute physical causes of disability. Methods: Five databases Medline, Embase, Web of Science, Scopus, and PsycINFO were searched for papers related to the validity, reliability, responsiveness, minimal clinically important difference or minimal detectable change of 12-item self-administered WHODAS 2.0. In order to avoid missing any potentially relevant studies, the search clauses were left as generic as possible and the refining search was conducted manually. As the review was focusing on chronic physical disorders and general adult population, major psychiatric diagnoses, acute traumas, other acute conditions (e.g., postpartum or pregnancy), hearing loss, progressive neurological disorders, and age <19 years were excluded. The relevancy of the studies was assessed by two independent reviewers. Results: The 14 out of 191 observational studies were considered relevant. The sample sizes varied from 80 up to 31,251 participants. Great diversity was observed in the participants' health problems. The Cronbach's alpha was high - up to 0.96. The correlations between WHODAS 2.0 and other disability scales were high. Substantial floor without ceiling effect was reported by two studies. Exploratory factor analysis resulted in a multidimensional structure - up to five factors. The discriminative ability and test-retest reliability of the scale was good. Conclusions: It seems, that the 12-item self-administered WHODAS 2.0 is internally consistent and a reliable scale demonstrating overall good correlation with other measures of disability. However, it appears that it is a multidimensional scale and its total score may represent different combinations of several contributing factors. Thus, the 12-item WHODAS 2.0 can be more reliable when creating a person's functional profile formed by the 12 individual item scores instead of a single total sum. IMPLICATIONS FOR REHABILITATION The 12-item self-administered WHODAS 2.0 is internally consistent and a reliable scale demonstrating overall good correlation with other measures of disability. It appears that it is a multidimensional scale and its total score may represent different combinations of several contributing factors. The 12-item WHODAS 2.0 can be more reliable when creating a person's functional profile formed by the 12 individual item scores instead of a single total sum.


Drug abuse prevention curricula in public and private schools in Indiana.
Recent research on drug education has focused on public schools. This study compares public school drug education programs with such efforts in private schools in one midwestern state. All schools in the state were asked to respond to a survey on curricula and training. This study reports on the types of prevention curricula used and at what grades, as well as the number of schools with trained staff. For almost every grade a higher percentage of public schools was found to be implementing drug education programs than private schools. This was manifested in public schools developing their own programs and/or purchasing commercially available drug curricula. Although an equal number of private and public schools have trained staff, less than half of all schools report a trained staff at each grade level. Directions for future research based on these results are suggested.


Minced cartilage without cell culture serves as an effective intraoperative cell source for cartilage repair.
Traumatic articular cartilage injuries heal poorly and may predispose patients to the early onset of osteoarthritis. One current treatment relies on surgical delivery of autologous chondrocytes that are prepared, prior to implantation, through ex vivo cell expansion of cartilage biopsy cells. The requirement for cell expansion, however, is both complex and expensive and has proven to be a major hurdle in achieving a widespread adoption of the treatment. This study presents evidence that autologous chondrocyte implantation can be delivered without requiring ex vivo cell expansion. The proposed improvement relies on mechanical fragmentation of cartilage tissue sufficient to mobilize embedded chondrocytes via increased tissue surface area. Our outgrowth study, which was used to demonstrate chondrocyte migration and growth, indicated that fragmented cartilage tissue is a rich source for chondrocyte redistribution. The chondrocytes outgrown into 3-D scaffolds also formed cartilage-like tissue when implanted in SCID mice. Direct treatment of full-thickness chondral defects in goats using cartilage fragments on a resorbable scaffold produced hyaline-like repair tissue at 6 months. Thus, delivery of chondrocytes in the form of cartilage tissue fragments in conjunction with appropriate polymeric scaffolds provides a novel intraoperative approach for cell-based cartilage repair.


New amine-functionalized cobalt cluster-based frameworks with open metal sites and suitable pore sizes: multipoint interactions enhanced CO2 sorption.
By employing the mixed-linker synthetic strategy, two new cobalt(II) cluster-based frameworks, [Co(2.5)(btc)(atz)2(Hatz)(DMF)]·2DMF (1) and [Co(2.5)(btc)(Hbtc)0.5(atz)(CH3CN)(H2O)]·H2O (2), constructed from the same initial materials Co(NO3)2, 1,3,5-benzenetricarboxylic acid (H3btc) and 3-amino-1,2,4-triazole (Hatz), have been synthesized by solvothermal reactions. Crystal-structure analyses demonstrated that 1 and 2 are three-dimensional (3D) porous frameworks based on Co-triazolate chain/layer and tricarboxylate pillars. Thermogravimetric analysis (TGA) and power X-ray diffraction (PXRD) measurements showed that both of them have high thermal stability and good water stability. Remarkably, both the structures of desolvated 1 and 2 contain suitable pore sizes and highly polar channel systems functionalized by amino groups, open metal sites, carbonyl or free carboxylic acid sites and uncoordinated triazolate nitrogen atoms on the pore surfaces, exhibiting multipoint interactions between CO2 molecules and frameworks, resulting in high CO2 uptake and selectivity for CO2 over N2.


Sildenafil (Viagra) evokes retinal arteriolar dilation: dual pathways via NOS activation and phosphodiesterase inhibition.
Sildenafil (Viagra; Pfizer, New York, NY), a selective phosphodiesterase type-5 (PDE5) inhibitor, is widely used to treat impotence by improving penile blood flow via elevation of cGMP. However, its effect on ocular circulation is controversial and whether retinal arterioles are responsive to this drug remains unclear. In this study, the direct reaction of retinal arterioles to sildenafil was examined and the signaling pathway underlying this vasomotor activity was probed. Retinal arterioles from porcine eyes were isolated, cannulated, and pressurized without flow. Diameter changes in response to sildenafil were recorded using videomicroscopic techniques. Retinal arterioles (67 +/- 2 microm) dilated dose dependently to sildenafil (1 ng/mL to 1 microg/mL). This dilation was inhibited by the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), the guanylyl cyclase inhibitor 1H- [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), the extracellular signal-regulated kinase (ERK) pathway inhibitor PD98059, the nonselective potassium channel blocker tetraethylammonium (TEA), and the selective adenosine triphosphate (ATP)-sensitive potassium (K(ATP)) channel blocker glibenclamide. The vasodilation elicited by the NO donor S-nitroso-N-acetylpenicillamine (SNAP) was inhibited by ODQ and TEA but was insensitive to PD98059. In the presence of L-NAME, the addition of SNAP (1 microM) produced modest vasodilation and the inhibited sildenafil response was subsequently restored. The restored dilation was insensitive to PD98059 but was blocked by TEA. Activation of NO synthase, through ERK signaling, leading to NO production and subsequent guanylyl cyclase activation and K(ATP) channel opening is the major vasodilatory pathway for sildenafil in retinal arterioles. Moreover, the elevated cGMP, from endogenous or exogenous NO, plays a permissive role for sildenafil to exert vasodilation through inhibition of the PDE5 pathway independent of ERK signaling.


The impact of fatigue on patient safety.
Modern medicine is founded on a culture of diligent, fatigued physicians. Fatigue is not desirable; however, the task of managing fatigue in health care professionals is complex and an ideal solution has not been described. Solutions need to integrate the immediate need for continued high-quality patient care, education of trainees, and the limited supply of health and human resources. Exploration of alternate scheduling models, broadened scope of practice, and new models of care delivery in demonstration projects or formal studies should be performed before widespread implementation. Appropriate evaluations are essential if well-meaning but larger scale errors in the name of patient safety are to be avoided.


Withdrawal of nursing services: strategies for survival.
In recent years, nursing strikes have become more common. Within the last decade, nurses in five of our ten provinces have all taken strike action. If this trend continues, more strikes could very likely occur. Administrative nurses provide direction and leadership for the nursing department. With the prospect of a withdrawal of nursing services, they are key players in preparing for the strike. They must ensure a standard of care during the strike, as well as facilitate the return of the unionized nurses following the strike. To assist administrative nurses who may some day be involved in a withdrawal of nursing services, this manuscript contains an account of the pre-strike preparations, putting these plans into action during the strike, and the immediate post-strike period at the Plains Health Centre, a tertiary care facility in Regina, Saskatchewan.


Hybrid photoacoustic fluorescence molecular tomography using finite-element-based inversion.
Improvements in fluorescence reconstruction when utilizing a hybrid photoacoustic (PAT) fluorescence molecular tomography (FMT) method to image optically heterogeneous media are studied and showcased. Quantitative optical absorption maps are retrieved using a normalized backprojection algorithm for PAT reconstruction. Consecutively, the reconstructed absorption distribution is employed into computing a diffusion-equation-based forward model for FMT using a finite-element solution. The potential promise of the suggested method is experimentally verified on tissue-mimicking fluorescent phantoms, where improvements in the quality of FMT reconstructions are observed when imaging at the presence of a large absorber.


Production of rabbit antibodies against the viral capsid antigen (VCA) of the Epstein-Barr virus (EBV).
Rabbits were immunized with nuclear or cytoplasmic extracts of the Epstein-Barr virus (EBV)-producing marmoset cell line B95-8. Following extensive absorption with human EBV-negative cells (HeP-2, Ramos and BJAB), sera were obtained that no longer reacted with cellular or serum proteins of human origin, but gave a single precipitin band with extracts of the human EBV-producing line, P3HR-1. Immunofluorescence tests performed with appropriate parallel human serum controls showed that the rabbit serum contained no activity against EBNA + EA - VCA - Raji cells, or against P3HR-1 virus superinfected, cytosine arabinoside-treated Raji cells that contained EBNA and EA, but not VCA. The sera gave a brilliant indirect immunofluorescence reaction with the virus-producing (EBNA+ EA+ VCA+) P3HR-1 lines. Two-color fluorescence tests, performed with a direct TRITC-labelled VCA conjugate and indirect FITC-staining with the rabbit serum, showed that the same cells reacted in both red and green fluorescence, confirming that the rabbit serum was directed specifically against some antigen formed in the virus-producer cells. Since the synthesis of the relevant antigen was prevented by cytosine arabinoside it cannot be EA and must be a late antigen. The morphology and localization of the antigen support the conclusion that the antigen is VCA or some part of the VCA complex.


[Molecular mechanisms of senescence in cell culture].
Clones of non-transformed human cells are mortal, that is, the cells divide only a limited number of times before they approach a state of proliferative senescence. This state has long been regarded as a cellular model of organismal aging or as aging in vitro because of its close relationship to the aging process of the donor of the cells. The underlying molecular mechanisms of this particular aging process have only been recently understood and are reviewed in the present paper. Cell division is accompanied by progressive telomere shortening, which is due both to properties of the replicative apparatus (the "end-replication problem") and to oxidative damage to telomere DNA under conventional cell culture conditions. If shortening of telomeres reaches a certain critical level, it is recognized as DNA damage by the cell's "guardian of the genome", the tumor suppressor p53. Stabilization of p53 activates the well characterized cell cycle checkpoint at the G1/S phase boundary and blocks the cell cycle irreversibly. Two recent results prove that telomere shortening is in fact the trigger of the checkpoint control in cellular senescence: First, acceleration of telomere shortening by increased oxidative stress results in accelerated proliferative senescence. Second, stabilization of telomere length, typically by activation of telomere, appears to be a necessary prerequisite for the immortalization of cells. Proliferative senescence, therefore, should be understood as an important means to counteract genetic instability and cancer.


SIRT1 suppresses colorectal cancer metastasis by transcriptional repression of miR-15b-5p.
The class III deacetylase sirtuin 1 (SIRT1), a member of the sirtuin family proteins, plays a key role in many types of cancers including colorectal cancer (CRC). Here we report that SIRT1 suppressed CRC metastasis in vitro and in vivo as a negative regulator for miR-15b-5p transcription. Mechanistically, SIRT1 impaired regulatory effects of activator protein (AP-1) on miR-15b-5p trans-activation through deacetylation of AP-1. Importantly, acyl-CoA oxidase 1 (ACOX1), a key enzyme of the fatty acid oxidation (FAO) pathway, was found as a direct target for miR-15b-5p. SIRT1 expression was positively correlated with ACOX1 expression in CRC cells and in xenografts. Moreover, ACOX1 overexpression attenuated the augmentation of migration and invasion of CRC cells by miR-15b-5p overexpression. In conclusion, our study demonstrated a functional role of the SIRT1/miR-15b-5p/ACOX1 axis in CRC metastasis and suggested a potential target for metastatic CRC therapy.


Importation of in vitro-produced Bubalus bubalis embryos from Italy into the United States: a case report.
On December 19, 2005, 14 in vitro-fertilized water buffalo (Bubalus bubalis) embryos, which had been cryopreserved by vitrification, were thawed and transferred into B. bubalis recipients in California. The embryos had been produced in Italy, following transvaginal oocyte pickup (TVOPU), with subsequent in vitro maturation, insemination, and culture. This case study relates our experience in meeting the regulatory criteria, established by the Animal Import/Export Office of the USDA-Animal and Plant Health Inspection Service (USDA-APHIS), in order to successfully import these embryos into the USA.


Effect of norfloxacin, trimethoprim-sulfamethoxazole and nitrofurantoin on fecal flora of women with recurrent urinary tract infections.
The effects of antibiotics used for prophylaxis in women with recurrent urinary tract infections (UTIs) on the aerobic intestinal flora were investigated. Twenty-one patients with recurrent UTIs were randomly divided into three groups. The patients of each group received monotherapy with oral norfloxacin, trimethoprim-sulfamethoxazole or nitrofurantoin for one month. Urine and stool quantitative aerobic cultures were performed before prophylaxis, 2 and 4 weeks after the initiation of therapy, and 2 weeks after antibiotics were discontinued. The gram-negative aerobic flora was strongly suppressed during the administration of norfloxacin and trimethoprim-sulfamethoxazole, while Enterococcus spp. were not affected. Resistant strains of Escherichia coli were detected in two patients, one in the norfloxacin and one in the trimethoprim-sulfamethoxazole group. The aerobic intestinal flora was not affected by nitrofurantoin. These findings help in the selection of the most appropriate antimicrobial agent for prophylaxis in recurrent UTIs, so as to reduce the possibility of emergence of resistant bacterial strains.


Atherosclerosis and secondary deep vein thrombosis: a difficult correlation.
The aim of this study was to verify the degree of atherosclerosis in a group of subjects affected by secondary deep vein thrombosis and in a matched control group. Sixty-three patients were studied. Of these, 19 were cases (mean age 62.2 +/- 1.2) and 16 were controls (mean age 59.3 +/- 2.7). Twenty-eight were excluded because they were affected by hyperhomocysteinemia. The arterial tree was examined by means of echo color Doppler, and the intima media thickness and the presence of and degree of plaques bilaterally in the carotid and femoral artery and abdominal aorta were measured with a computerized method. No difference was found in the 2 groups as concerns intima media thickness or plaques in the arterial district explored. Secondary deep vein thrombosis is not a greater risk factor for atherosclerosis.


Immunohistochemical analysis of cyclic AMP response element binding protein phosphorylation in focal cerebral ischemia in rats.
Phosphorylation of cyclic AMP response element binding protein (CREB) is one of the most important mechanisms controlling various gene transcriptions. In the present study, the phosphorylation of CREB was examined immunohistochemically at 24 h of recirculation following 1.5 h of middle cerebral artery occlusion (MCAO) in rats. MCAO was induced by the intraluminal suture method. The infarct core revealed a significant reduction in the number of immunoreactive cells with the anti-phosphorylated CREB and with the anti-CREB antibody, which binds to both unphosphorylated and phosphorylated CREB. In contrast, the peri-infarct area exhibited a marked increase in the number of immunopositive cells as well as in the intensity of nuclear staining with each antibody, so that almost all of the cells expressing CREB demonstrated phosphorylation of CREB. On the other hand, about half of the CREB immunopositive cells reacted weakly with the anti-phosphorylated CREB antibody in the sham group. These findings indicated that the expression as well as phosphorylation of CREB protein was significantly activated in the regions surrounding the infarct area. Since phosphorylation of CREB has recently been implicated in signal transductions that promote the survival and differentiation of neurons, the present data suggest that tissue repair mechanisms may be markedly activated in the peri-infarct area.


Unraveling the genetic basis of fast l-arabinose consumption on top of recombinant xylose-fermenting Saccharomyces cerevisiae.
One major challenge in the bioconversion of lignocelluloses into ethanol is to develop Saccharomyces cerevisiae strains that can utilize all available sugars in biomass hydrolysates, especially the d-xylose and l-arabinose that cannot be fermented by the S. cerevisiae strain naturally. Here, we integrated an l-arabinose utilization cassette (AUC) into the genome of an efficient d-xylose fermenting industrial diploid S. cerevisiae strain CIBTS0735 to make strain CIBTS1972. After evolving on arabinose, CIBTS1974 with excellent fermentation capacity was obtained. A comparison between genome sequences of strains CIBTS1974 and CIBTS1972 revealed that the copy number of the AUC had increased from 1 to 12. We then constructed the AUC null-mutant CIBTS1975 and gradually rescued the l-arabinose utilization defect by integrating AUC iteratively. On the other hand, the parental strain CIBTS0735 was able to acquire the same performance as CIBTS1974 by the direct introduction of 12 copies of the AUC; the performance was further improved by adding two more copies. Besides, we found that not the two transporters present in the AUC were both needed during l-arabinose utilization, GAL2 was necessary and STP2 was not essential. We have described for the first time that a high copy number of AUC is sufficient for the strain to metabolize l-arabinose efficiently independent of evolution.


Atrial fibrillation in the elderly.
Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and its prevalence is increasing with age. With aging of the population treatment of atrial fibrillation especially in elderly population is a growing task for all medical staff working with elderly patients. Treatment of atrial fibrillation especially in elderly patients has to focus on prevention of thromboembolism as well as symptom relief with rate or rhythm control. This review article will focus on medical and non-pharmacological treatment options for the treatment of atrial fibrillation in elderly patients.