Atomic force microscopy characterization of palmitoylceramide and cholesterol effects on phospholipid bilayers: a topographic and nanomechanical study.
Supported planar bilayers (SPBs) on mica substrates have been studied at 23 °C under atomic force microscopy (AFM)-based surface topography and force spectroscopy with two main objectives: (i) to characterize palmitoylceramide (pCer)-induced gel (Lβ) domains in binary mixtures with either its sphingolipid relative palmitoylsphingomyelin (pSM) or the glycerophospholipid dipalmitoylphosphorylcholine (DPPC) and (ii) to evaluate effects of incorporating cholesterol (Chol) into the previous mixtures in terms of Cer and Chol cooperation for the generation of lamellar gel (Lβ) phases of ternary composition. Binary phospholipid/pCer mixtures at XpCer < 0.33 promote the generation of laterally segregated micron-sized pCer-rich domains. Their analysis at different phospholipid/pCer ratios, by means of domain thickness, roughness, and mechanical resistance to tip piercing, reveals unvarying AFM-derived features over increasing pCer concentrations. These results suggest that the domains grow in size with increasing pCer concentrations while keeping a constant phospholipid/pCer stoichiometry. Moreover, the data show important differences between pCer interactions with pSM or DPPC. Gel domains generated in pSM/pCer bilayers are thinner than the pSM-rich surrounding phase, while the opposite is observed in DPPC/pCer mixtures. Furthermore, a higher breakthrough force is observed for pSM/pCer as compared to DPPC/pCer domains, which can be associated with the preferential pCer interaction with its sphingolipid relative pSM. Cholesterol incorporation into both binary mixtures at a high Chol and pCer ratio abolishes any phospholipid/pCer binary domains. Bilayers with properties different from any of the pure or binary samples are observed instead. The data support no displacement of Chol by pCer or vice versa under these conditions, but rather a preferential interaction between the two hydrophobic lipids.


Cysteine-Based Protein Adduction by Epoxide-Derived Metabolite(s) of Benzbromarone.
Benzbromarone (BBR) is a therapeutically useful uricosuric agent but can also cause acute liver injury. The hepatotoxicity of BBR is suggested to be associated with its metabolic activation. Our recent metabolic study demonstrated that BBR was metabolized to epoxide intermediate(s) by cytochrome P450 3A, and the intermediate(s) was reactive to N-acetylcysteine. The objectives of the present study were to determine the chemical identity of the interaction of protein with the epoxide intermediate(s) of BBR and to define the association of the protein modification with hepatotoxicity induced by BBR. Microsomal incubation study showed that the reactive intermediate(s) covalently modified microsomal protein at cysteine residues. Such adduction was also observed in hepatic protein obtained from liver of mice given BBR. The protein covalent binding occurred in time- and dose-dependent manners. Pretreatment with ketoconazole attenuated BBR-induced protein modification and hepatotoxicity, while pretreatment with dexamethasone or buthionine sulfoximine potentiated the protein adduction and hepatotoxicity induced by BBR. A good correlation was observed between BBR-induced hepatotoxicity and the epoxide-derived hepatic protein modification in mice. The present study provided in-depth mechanistic insight into BBR-induced hepatotoxicity.


HIF-1α and HIF-2α correlate with migration and invasion in gastric cancer.
Hypoxia-inducible factor-1 (HIF-1) is a major determinant of invasion and metastasis in several tumor types. We previously reported that HIF-1α contributed to multidrug resistance in gastric cancer. However, the role of HIF-2α on the progression of gastric cancer is seldom reported. In this study, we first examined the possible role of HIF-1α and HIF-2α in the process of invasiveness and metastasis of gastric cancer, using immunohistochemistry of 80 gastric cancer tissues, western blot and real-time PCR of 8 fresh gastric cancer tissues. The results showed that HIF-1α and HIF-2α were significantly correlated with the clinical stage and were highly expressed in metastatic gastric cancers compared to nonmetastatic ones. Western blot analysis revealed that hypoxia (1% O₂, 8 h) induced HIF-1α and HIF-2α expression in different gastric cancer cell lines, including SGC7901, AGS, MGC803 and MKN45. Adhesion and invasion assays found that hypoxia caused an increase in adhesive and invasive abilities of gastric cancer cells. Small interfering (si) RNA against HIF-1α and HIF-2α in SGC7901 cells significantly inhibited hypoxia-induced adhesive and invasive abilities. Finally, the JNK inhibitor SP 600125 abolished hypoxia-induced HIF-1α and HIF-2α expression, and inhibited the adhesive and invasive abilities of gastric cancer cells exposed to hypoxia in a dose-dependent manner. Taken together, the present work suggested that HIF-1α and HIF-2α were involved in metastasis and invasion of gastric cancer cells under hypoxia, with the involvement of JNK signal pathway.


Mental health of intermarried immigrant women and their children in South Korea.
This study was aimed to examine the mental health status of immigrant women through international marriages and its effects on emotional and behavioral problems of children. Seventy-four intermarried immigrant women and 86 native Korean women were enrolled from the same district of Seoul metropolitan area. The mental health problems of study participants and their children were examined. Increased risks of having a higher level of anxiety were observed in immigrant women compared to native Korean women. Children of the immigrant group were likely to have more internalizing and externalizing behavioral symptoms than those of the native group. This pattern was more apparent in children of mothers with higher levels of anxiety. The present study found a higher risk for mild anxiety in intermarried immigrants than in native Korean women. Furthermore, considering that immigrant children seemed to have more emotional and behavioral problems relative to native children if their mothers have higher levels of anxiety, special attention should be paid to prevention and early intervention for mental health problems of intermarried immigrant women.


Heterodimerization of c-Jun with ATF-2 and c-Fos is required for positive and negative regulation of the human urokinase enhancer.
Dimerization plays a pivotal role in modulating the activity of the c-Jun proto-oncogene product. Heterodimerization with activating transcription factor-2 (ATF-2) alters the DNA-binding specificity of c-Jun, allowing its targeting to several cAMP responsive element (CRE)-related sequences, which control a subset of AP-1-responsive genes. Here we show that a c-Jun/ATF-2 heterodimer binds to the AP-1 site (uPA 5'-TRE) essential for the activity of the human urokinase enhancer, conferring on this element several distinctive regulatory properties. The c-Jun/ATF-2 heterodimer was identified by binding competition assays, u.v. cross linking, and monospecific antibodies. In vitro binding studies revealed that the uPA 5'-TRE sequence is recognized by the cyclic AMP-unresponsive ATF-2 factor, but not by the cyclic AMP-inducible CREB. In addition, in vivo studies suggest that ATF-2 can mediate, at the same time, the activation of the c-Jun/ATF-2 site and the repression of the canonical collagenase AP-1 site. We report that heterodimerization with c-Fos does not increase the binding of c-Jun to the uPA 5'-TRE, in contrast to the increased binding at a consensus AP-1 site. Our data further suggest that c-Fos can act as a repressor of the c-Jun/ATF-2 binding site, revealing an important functional difference, with respect to canonical AP-1 elements.


Bayesian Inference for Neighborhood Filters With Application in Denoising.
Range-weighted neighborhood filters are useful and popular for their edge-preserving property and simplicity, but they are originally proposed as intuitive tools. Previous works needed to connect them to other tools or models for indirect property reasoning or parameter estimation. In this paper, we introduce a unified empirical Bayesian framework to do both directly. A neighborhood noise model is proposed to reason and infer the Yaroslavsky, bilateral, and modified non-local means filters by joint maximum a posteriori and maximum likelihood estimation. Then, the essential parameter, range variance, can be estimated via model fitting to the empirical distribution of an observable chi scale mixture variable. An algorithm based on expectation-maximization and quasi-Newton optimization is devised to perform the model fitting efficiently. Finally, we apply this framework to the problem of color-image denoising. A recursive fitting and filtering scheme is proposed to improve the image quality. Extensive experiments are performed for a variety of configurations, including different kernel functions, filter types and support sizes, color channel numbers, and noise types. The results show that the proposed framework can fit noisy images well and the range variance can be estimated successfully and efficiently.


Antitumor polysaccharides from a Chinese mushroom, "yuhuangmo," the fruiting body of Pleurotus citrinopileatus.
A water-soluble polysaccharide, FI, and water-insoluble polysaccharides, FII and FIII were extracted from Pleurotus citrinopileatus mushrooms. After fractionation, the antitumor activity of the fractions against Sarcoma 180 implanted in mice were examined, and the following active polysaccharides were obtained: Water-soluble polysaccharides FIo: A heteropolysaccharide containing 9.8% protein, and composed of glucose, mannose, arabinose, and galactose. FA-2-b-beta: A glycoprotein consisting of glycan:protein = 40:60 w/w, with the glyco-chain composed of glucose, xylose, mannose, galactose, and fucose. FA-3: A glycoprotein consisting of glycan:protein = 50:50 w/w, and glycan moiety consisting of glucose, galactose, xylose, mannose, and fucose. Water-insoluble polysaccharides. FIII-1: Protein-containing beta-D-glucans, FIII-1-a, and -b were obtained from FIII-1 by gel filtration, composed of glucan:protein = 80:20, and 68:32 w/w, respectively. The glucan moieties of both were almost all (1-->3)-beta-D-glucan, and their molecular weights were 68 x 10(4) and 40 x 10(4). FIII-2: Protein-containing beta-D-glucans, FIII-2-a, and -b, were obtained from FIII-2 by gel filtration, with molecular weights 190 x 10(4) and 120 x 10(4), respectively. Both were almost all composed of glucan:protein = 87:13 w/w. Both glucan moieties were mainly (1-->3)-beta-D-glucan.


Anxiety and self-dissatisfaction in epileptic children.
Twenty epileptic children were compared with regard to their anxiety levels and self-concept measures, to two control groups: (a) non-handicapped children, (b) learning-disabled children. The epileptic children demonstrated higher levels of trait anxiety and dissatisfaction with themselves, as compared to each and both of the control groups. The results stressed the critical value of psychological consultation for the epileptic children and their parents.


Changes in ovarian and pituitary function in non-pregnant women during the infusion of prostaglandin F2alpha.
The concentrations of prostaglandin F2alpha, progesterone, oestradiol, LH and cortisol have been determined in serial samples of peripheral venous plasma, before, during and after, the intravenous administration of prostaglandin F2alpha (50 mug/min, for 5 h) to 8 women in the luteal phase of the ovarian cycle. The results show that the mean concentrations of prostaglandin F2alpha and cortisol increased during the infusion by factors of 8.8 and 2.6 respectively. The highest levels of cortisol occurred after 3 h, and were significantly different (P less than 0.0005, Student's t-test) from those obtained before the infusion. There were progressive decreases in the concentrations of LH and progesterone. After 3 h the values for LH were significantly lower (P less than 0.0025) than those before the administration of prostaglandin F2alpha, and the lowest mean value was 26% of the control. The values for progesterone were significantly lower (P less than 0.025) after 5 h, and the mean value at this time was 38% of the control. The pattern of the mean concentration for oestradiol was similar to that for progesterone. The levels of all 5 compounds had returned to normal within 1 h after completion of the infusion. The findings are discussed.


Ischaemic deterioration following aneurysmal subarachnoid haemorrhage: definition by clinical criteria.
Ischaemia following surgery for aneurysmal subarachnoid haemorrhage (SAH) was defined by clinical criteria. Two distinct groups of patient developing ischaemia were identified: (1) patients recovering from anaesthesia with a new deficit or (2) patients developing 'true' delayed ischaemia. The only significant difference between the groups was an excess of pre-existing vascular disease in the former group. The eventual outcome for patients developing an immediate postoperative deficit was not significantly different from the delayed ischaemia group. The 6-month outcome for patients without clinical ischaemia in the first 10 postoperative days was excellent. The potentially different aetiology of ischaemia in the two subgroups with clinical ischaemia identified in this study should be considered during the planning and in particular the statistical design of trials testing new treatments for clinical ischaemia following aneurysmal SAH.


Prednisolone absorption in coeliac disease.
The pharmacokinetic disposition of prednisolone was studied following oral administration (10 mg) to eight patients with untreated coeliac disease of mild or moderate severity, seven coeliac patients on a strict gluten-free diet and ten normal subjects. No significant differences were shown in the peak prednisolone level, the time of the peak level, the area under the concentration versus time plot, the plasma half-life and the 24 h urinary recovery of prednisolone in the three subject groups. There was however considerable variability within each group. It is concluded that the presence of coeliac disease does not significantly alter the absorption or elimination of prednisolone.


Testing declarative memory in laboratory rats and mice using the nonconditioned social discrimination procedure.
Testing declarative memory in laboratory rodents can provide insights into the fundamental mechanisms underlying this type of learning and memory processing, and these insights are likely to be applicable to humans. Here we provide a detailed description of the social discrimination procedure used to investigate recognition memory in rats and mice, as established during the last 20 years in our laboratory. The test is based on the use of olfactory signals for social communication in rodents; this involves a direct encounter between conspecifics, during which the investigatory behavior of the experimental subject serves as an index for learning and memory performance. The procedure is inexpensive, fast and very reliable, but it requires well-trained human observers. We include recent modifications to the procedure that allow memory extinction to be investigated by retroactive and proactive interference, and that enable the dissociated analysis of the central nervous processing of the volatile fraction of an individual's olfactory signature. Depending on the memory retention interval under study (short-term memory, intermediate-term memory, long-term memory or long-lasting memory), the protocol takes ~10 min or up to several days to complete.


Development of quality control preparations for immunocytochemical assessment of urokinase-type plasminogen activator.
Quality control of immunochemical and/or immunocytochemical analyses warrants constant reproducibility and reliability of assay performance. In this respect, stable reference preparations containing known quantities of the components to be assessed may serve purposes in the quality assessment of antigen expression levels, including those of the plasminogen activation system. Quality control preparations for the immunocytochemical assessment of urokinase-type plasminogen activator (uPA) were developed using different combinations of cultured cell lines (BLM and IF6), each expressing immunochemically well-defined (by enzyme-linked immunosorbent assay[ELISA]) amounts of the respective component. Cytospins and frozen sections cut from sucrose/Tissue-Tek blocks containing these cell lines demonstrated stable and homogeneous expression of uPA. An excellent correlation was found between the immunocytochemical staining results and the data obtained by ELISA. Because these cell lines are available in practically unlimited quantities, large numbers of nearly identical quality control preparations can be made over a long period of time. Therefore, the incorporation of (combinations of) cell lines in cytospins or sucrose/Tissue-Tek blocks represents a simple model system in establishing quality control preparations for immunocytochemical assessment of components of the plasminogen activator system.


Rheumatoid factors in systemic lupus erythematosus: association with clinical and laboratory parameters. SLE study group.
The prevalence and clinical and laboratory associations of IgM, IgG and IgA rheumatoid factors (RF) were determined in 352 patients with systemic lupus erythematosus (SLE). IgM, IgG, and IgA class RF were detected in 17.9%, 20.5%, and 20.5% of the sera, respectively. RF were associated with sicca syndrome, hypergammaglobulinemia, high titer of antinuclear antibodies, anemia, SSA- and SSB-antibodies, and with the presence of HLA-DR3. RF correlated negatively with nephritis and livedo racemosa. Moreover, we observed an association of RF and parameters of inflammatory activity such as elevated erythrocyte sedimentation rate (ESR) and leukopenia. Analysis of immunoglobulin classes revealed that laboratory parameters of inflammatory activity, SSA- and SSB-antibodies and HLA-DR3 correlated with IgA RF only. IgA RF define a subgroup of SLE patients characterized by distinct autoimmune phenomena and high disease activity in the absence of nephritis.


Structural basis of calcium and galactose recognition by the lectin PA-IL of Pseudomonas aeruginosa.
The structure of the tetrameric Pseudomonas aeruginosa lectin I (PA-IL) in complex with galactose and calcium was determined at 1.6 A resolution, and the native protein was solved at 2.4 A resolution. Each monomer adopts a beta-sandwich fold with ligand binding site at the apex. All galactose hydroxyl groups, except O1, are involved in a hydrogen bond network with the protein and O3 and O4 also participate in the co-ordination of the calcium ion. The stereochemistry of calcium galactose binding is reminiscent of that observed in some animal C-type lectins. The structure of the complex provides a framework for future design of anti-bacterial compounds.


[Effect of danhong huayu oral liquid combined with insulin on prevention and progression of early diabetic cardiomyopathy in rats].
To investigate effects of Danhong Huayu Koufuye (DHK), insulin and their combination on diabetic cardiomyopathy (DC) in streptozotocin (STZ, 50 mg/kg, ip)-induced diabetic rats. Rats were divided into five groups: normal control, diabetic treated with vehicle, insulin, DHK, and DHK plus insulin. The animals were treated once daily for 15 weeks starting one week after STZ injection. The combination of DHK with insulin significantly reduced cardiac index (P < 0.05), serum LDH (P < 0.05), AST(P < 0.05), ALT(P < 0.05) and HDL-C (P < 0.05) level, and promoted pancreatic and cardiac morphological changes as compared with the model group. It is suggested that DHK may be a valuable adjuvant therapy for DC.


Resident-Led Palliative Care Education Project.
Despite the growth of palliative medicine, 39% of hospitals do not have palliative care teams for consultation or to provide resident education. We examined the impact of resident-led education in palliative care principles on attitudes toward and comfort with palliative medicine and end-of-life care among internal medicine residents. An educational module designed by the authors was presented to other internal medicine residents in the program. Pre- and post-intervention survey data measuring residents' agreement with various statements regarding palliative medicine and end-of-life care were analyzed. Residents' agreement with various statements regarding palliative medicine and end-of-life care on a 5-point Likert scale was analyzed. Following the intervention, participants reported improved comfort with general knowledge of palliative medicine (p < 0.01), specific resources available to patients (p < 0.001), and explaining the difference between palliative care and end-of-life care (p < 0.001). In each of the seven specific domains of palliative medicine covered in the educational session, residents reported a statistically significant increase in comfort in all of the areas addressed (p < 0.05). This study demonstrates that a resident-led curriculum in palliative medicine can improve resident comfort within this still-under-represented area of medicine.


Paclitaxel-loaded crosslinked hyaluronic acid films for the prevention of postsurgical adhesions.
Post surgical adhesion formation results in significant morbidity for surgical patients. The purpose of this study was to investigate the use of paclitaxel (PTX) as an inhibitor of adhesion formation in rats and to design and characterize a controlled release film formulation of the drug for application to exposed surgical sites. The rat cecal side wall abrasion model was used to investigate the anti-adhesion properties of PTX. The drug was administered by either intraperitoneal injection (i.p.), as the cremophor formulation (Taxol) or by application of carbodiimide crosslinked hyaluronic acid (HA) films containing PTX. The HA films were also characterized by measurements of elasticity, degree of swelling in water and drug release rates. Taxol administered by i.p. injection at 4 mg/kg on a daily basis for between 3 and 5 days resulted in a significant reduction in adhesion formation. All animals in the control group (n = 10) had some form of adhesion following abrasion whereas the percent of animals without adhesions significantly increased and the mean incidence of adhesion formation decreased in the three Taxol treated groups. The application of 5% PTX loaded HA films had a similar significant effect in increasing both the % of animals without adhesions and in reducing the mean incidence of adhesions. Paclitaxel is an effective inhibitor of adhesion formation in rats. HA crosslinked with 2 mM water soluble carbodiimide and containing 10% glycerol and 5% PTX are flexible, mucoadhesive, biocompatible controlled release films suitable for application to surgical sites for the prevention of adhesion formation.


Bone mineral densitometry in patients on hemodialysis: difference between genders and what to measure.
Chronic kidney disease (CKD) and osteoporosis are important health problems. There is an interrelationship between osteoporosis and CKD. Bone densitometry is the "gold" standard in the diagnosis of osteoporosis. Unfortunately, there are some problems with the interpretation of bone densitometry in CKD patients. The goal of this study was to determine bone mineral density (BMD) in CKD patients, to assess the difference between genders and different sites of bone densitometry correlation between BMD and laboratory parameters, and to assess the most optimal measuring site. We studied 134 hemodialysis (HD) patients (62 females, 72 males). The mean age was 56.4+/-12.4 years and the mean duration of HD was 54.4+/-60 months. BMD of the lumbar spine (posterior-anterior projection and lateral projection), hip (femoral neck, trochanter, intertrochanter, total femur, the Ward's Triangle), and forearm (ultradistal (UD), middistal (MID), distal third portion, and total forearm) was measured using dual X-ray absorptiometry (DXA) (Hologic Delphi apparatus). Values were expressed as BMD, T-score, and Z-score. Females had lower values of BMD in all measurement points. There were no significant differences in T- and Z-scores of forearm between males and females. Age was in a positive correlation with lumbar spine BMD in males and females. There was a negative correlation with neck and forearm BMD in both groups. Serum parathyroid hormone (PTH) was also in negative correlation with hip and forearm BMD in both groups. The best correlation of BMD in different sites was between forearm and neck. BMD data in CKD patients should be interpreted with caution and appendicular skeletal sites should be included in the evaluation.


β-Amino acid catalyzed asymmetric Michael additions: design of organocatalysts with catalytic acid/base dyad inspired by serine proteases.
A new type of chiral β-amino acid catalyst has been computationally designed, mimicking the enzyme catalysis of serine proteases. Our catalyst approach is based on the bioinspired catalytic acid/base dyad, namely, a carboxyl and imidazole pair. DFT calculations predict that this designed organocatalyst catalyzes Michael additions of aldehydes to nitroalkenes with excellent enantioselectivities and remarkably high anti diastereoselectivities. The unusual stacked geometry of the enamine intermediate, hydrogen bonding network, and the adoption of an exo transition state are the keys to understand the stereoselectivity.


Craniomandibular disorders in children--a critical review of the literature.
Reports of prevalences of craniomandibular disorders (CMD) in children and adolescents differ markedly from one study to another. Therefore it is almost impossible to get a clear picture of the magnitude of the disorder in a child population. The aim of this study was to critically compare and analyse the results presented in recent publications in order to get a comprehensive view of the prevalence of CMD in children and adolescents. Fourty epidemiological studies on CMD symptomatology and headache in the age group three to 25 years were included. Eight symptoms and eight signs were chosen for further comparisons and analyses. The prevalences of these signs and symptoms were evaluated and classified according to a set of criteria, in either frequency of "greater clinical relevance" or frequency of "lesser clinical relevance". The evaluation was based on the presented examination methods, definitions and symptom criteria used in the studies. The reported frequencies for the sixteen signs and symptoms are presented graphically. The diagram clearly show great variation in the reported frequencies for most signs and symptoms. If only reported frequencies of "greater clinical relevance" are considered, these variations are less pronounced but still considerable. One major reason for this is inter- and intraindividual variation among the examiners. Another plausible and often overlooked reason is the fact that examination methods designed for adults uncritically have been applied on children without consideration of age and cognitive development of the child.


The effects of fungal infection and physiological condition on the locomotory behaviour of the ant Myrmica scabrinodis.
Parasite infection often results in alterations in host behaviour. These changes vary greatly in their magnitude, from slight shifts in the time spent by the host performing a given activity to the appearance of novel behaviours. The effects of parasites can differ with the age and the physiological condition of the host. Rickia wasmannii is an ectoparasitic fungal symbiont in Myrmica ants that covers the whole body surface of the host and reduces its lifespan. The fungus is present in both young and old individuals, making it an optimal subject for the study of age-related parasitic effects. We tested the effect of fungal infection on the locomotory activity of the Myrmica scabrinodis ant in different age categories. The fat content of workers was measured as a proxy for their physiological status. Based on our findings, old workers bore more thalli and were leaner than young individuals, while they tended to move at higher speeds and with a lower degree of meandering. Young individuals covered smaller distances, at slower speeds and with a higher degree of meandering. Contrary to our expectations, the infection intensity of R. wasmannii affected neither the fat content nor the locomotory activity of ant workers. However, the two age classes seem to have different strategies with regards to the relationship between fat content and distance covered. Our results suggest that characteristics of locomotory activity differ between the age classes in many respects, and are also influenced by their physiological status, but parasitism by R. wasmannii does not seem to have a straightforward effect on any of the variables studied.


Correlation of Synovial Fluid Biomarkers With Cartilage Pathology and Associated Outcomes in Knee Arthroscopy.
To correlate the intraoperative concentrations of 20 synovial fluid biomarkers with preoperative symptoms, intraoperative findings, and postoperative outcomes in patients undergoing knee arthroscopy, with comparisons made to samples obtained from asymptomatic knees. Synovial fluid samples were obtained from 81 patients undergoing knee arthroscopy meeting the inclusion criteria, which included 70 samples from operative knees and 32 samples from contralateral knees. Preoperatively, baseline data obtained from clinical questionnaires including a visual analog scale (VAS) score, the Lysholm score, and the Knee Injury and Osteoarthritis Outcome Score-Physical Function Short Form were recorded. Synovial fluid was collected from both the operative knee and asymptomatic contralateral knee. Synovial fluid was stored with a protease inhibitor at -80°C until analysis. Intraoperative findings, procedures performed, and International Cartilage Repair Society (ICRS) cartilage status scores in all operative knees were documented. The concentrations of the following 20 biomarkers were measured using a multiplex magnetic bead immunoassay: matrix metalloproteinase (MMP) 3; MMP-13; tissue inhibitor of metalloproteinase (TIMP) 1; TIMP-2; TIMP-3; TIMP-4; fibroblast growth factor 2; eotaxin; interferon γ; interleukin (IL) 10; platelet-derived growth factor BB; IL-1 receptor antagonist; IL-1β; IL-6; monocyte chemotactic protein 1 (MCP-1); macrophage inflammatory protein 1α; macrophage inflammatory protein 1β; RANTES (regulated upon activation, normal T cell expressed and secreted); tumor necrosis factor α; and vascular endothelial growth factor. Clinical outcome scores were obtained in 83% of patients at a mean of 17 months' follow-up postoperatively. Analysis of variance and Pearson correlation analysis were performed to determine statistical significance between preoperative data, intraoperative findings, postoperative outcomes, and synovial fluid biomarker concentrations compared with asymptomatic contralateral knees. Analysis was performed on 70 operative and 32 contralateral samples. There were strong positive correlations between ICRS score and age, symptom duration, VAS score, and Knee Injury and Osteoarthritis Outcome Score-Physical Function Short Form. A strong positive correlation was found between MCP-1 and IL-6 concentrations, intraoperative ICRS score, and continued pain at the time of final follow-up. MCP-1 and IL-6 were the strongest predictors of severe cartilage lesions, whereas IL-1 receptor antagonist was inversely related. MMP-3 levels were consistently elevated in all operative samples and directly correlated to increased preoperative VAS scores. RANTES, vascular endothelial growth factor, and platelet-derived growth factor BB were the strongest predictors of postoperative improvement at final follow-up regardless of injury and cartilage status. Synovial fluid biomarkers have the capacity to reflect the intra-articular environment before surgery and potentially predict postoperative clinical outcomes. Recognition of key molecular players may yield future therapeutic targets, and large clinical trials exploring these discoveries are anticipated. Level III, therapeutic case-control study.


Fractionation of plasma proteins using dye-ligand chromatography: elution profiles on immobilized green TSK-AF.
The fractionation of human plasma by chromatography on immobilized Green TSK-AF was assessed by immunological analysis of the elution profiles of 27 different plasma proteins. A three-step procedure was used to elute proteins from the column. First a low-molarity buffer (30 mM sodium phosphate, pH 7.0, I = 0.053) was applied; then a linear salt gradient (0-1.0 M NaCl in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M NaCl. Tightly bound proteins were finally stripped with 0.5 M NH4SCN. The elution profile of the proteins using this procedure appears to be very reproducible. Comparison with the profile obtained upon chromatography on Cibacron Blue 3GA [Gianazza, E. and Arnaud, P. (1982) Biochem. J. 201, 129-136] indicates significant differences between the binding properties of the two gels. These differences can be used to design a "tandem-chromatography" system which provides an efficient means for the separation of several plasma proteins.


Using Multitheory Model to Predict Low-Salt Intake Among Nigerian Adults Living With Hypertension.
One of the public health recommendations for the management of hypertension is the reduction of sodium/salt intake. The purpose of this study was to use the novel multitheory model of health behavior change to predict the initiation and maintenance of low-salt intake among adult Nigerian hypertensives. A quantitative cross-sectional design utilizing a convenience sample of 149 consenting Nigerian adults living with hypertension were self-administered a valid and reliable 39-item instrument. Multivariate regression analysis revealed 40.6% of the variance in initiating the consumption of low-salt diets explained by advantages outweighing disadvantages, behavioral confidence, and changes in physical environment. About 41.8% of the variance to sustain the intake of low-salt diet was explained by emotional transformation, practice for change, and changes in social environment. The results justified the predictive role of multitheory model and adequacy of its utility to build evidence-based health education interventions for hypertension in Nigeria.


Daytime leg pain after overnight continuous positive airway pressure titration.
We report a case that showed transitional, severe emergent periodic limb movements during sleep (PLMS) in the first continuous positive airway pressure (CPAP) titration night and mild PLMS in the 3rd and 20th nights of CPAP titration in a 56-year-old woman after she was found to have severe obstructive sleep apnea (OSA). Severe PLMS in the initial CPAP night unexpectedly resulted in acute leg muscle soreness in the second morning. The case suggests that CPAP therapy in patients who suffer from severe OSA may provide a unique opportunity to look into the pathological mechanism of PLMS.


Effect of physostigmine and atropine on acetylcholine turnover in mouse brain.
The effect of physostigmine salicylate (0.5 mg/kg, i.p.) alone and in combination with atropine sulfate (25 mg/kg, i.p.) on levels of acetylcholine (ACh) and choline (Ch) and turnover of ACh has been studied in whole brain and striatum of mice. The animals were killed by focussed microwave irradiation and the turnover of ACh was studied after i.v. injection of deuterium labelled Ch by employing mass fragmentography. Physostigmine increased the levels of ACh in whole brain from 24.5--28.0 nmol/g(P less than 0.001) whereas there was no significant increase in striatum. The levels of Ch were also increased. The turnover rate of ACh was decreased in whole brain from 15.4 to 8.4 and in striatum from 52.9 to 24.4 nmol/g . min. Physostigmine given before or after atropine did not completely block the ACh lowering effect of atropine. When atropine was given before physostigmine the turnover rate of ACh in whole brain was increased to 24.2 nmoles/g . min. The results seem to indicate that there is no clear cut relation between the turnover rate and level of ACh in vivo. The increase of the turnover rate induced by atropine is masked unless a cholinesterase inhibitor is given to protect the newly synthesized labelled ACh released by atropine.


Targeted therapies for renal cell carcinoma.
The management of patients with metastatic renal cell carcinoma (RCC) has changed dramatically over the past few years. Nephrectomy remains an important intervention for localized RCC but systemic therapy is the mainstay of treatment for patients who relapse after surgery or who have metastatic RCC. Before 2005, medical therapies for RCC were limited to cytokine therapies, which are very toxic and benefit only a small percentage of patients. In 2017, therapeutic agents now include kinase and immune checkpoint inhibitors. Contemporary research with these agents is now focusing on combinatorial and perioperative therapy. The field is now faced with the evolving challenge of how to select the best therapy for each patient during their natural history of disease, which has created a strong interest in modern sequencing and molecular approaches to identify biomarkers to personalize treatments. New therapeutic agents and approaches are associated with different toxicities and financial burdens, which require consideration of value by measuring clinical benefit, toxicity, and the cost of each drug with an organized framework. In this Review, we discuss the mechanisms underlying RCC and how improved molecular understanding helped the development of therapies, as well as biomarkers of response to treatment. We also discuss the value of these agents and their impact on personalization of therapy and drug development for RCC.


Lithium 2,2,6,6-tetramethylpiperidide-mediated alpha- and beta-lithiations of epoxides: solvent-dependent mechanisms.
Lithium 2,2,6,6-tetramethylpiperidide (LiTMP)-mediated alpha- and beta-lithiations of epoxides are described. LiTMP displays a markedly higher reactivity than does lithium diisopropylamide, consistent with literature reports. Detailed rate studies of LiTMP/THF and LiTMP/Me(2)NEt mixtures reveal similar rates but significant mechanistic differences. LiTMP-mediated alpha-lithiation of cis-cyclooctene oxide with subsequent oxacarbenoid formation and transannular C-H insertion proceeds via monosolvated dimers in both THF and Me(2)NEt. LiTMP-mediated beta-lithiation of 2,3-dimethyl-2-butene oxide affords the corresponding allylic alcohol via a monosolvated monomer in THF and a monosolvated dimer in Me(2)NEt. We discuss how the solvent-dependent aggregation of LiTMP markedly influences the rate profile. The reaction transition structures are examined with density functional computations.


Hydrocortisone has a biphasic effect on rat gastric mucosal prostaglandin generation in vivo: inhibition at low doses, stimulation at high doses.
To determine the effect of different doses of hydrocortisone sodium succinate (HC) on rat gastric mucosal prostaglandin synthesis, two experiments were performed. In the first experiment, 20 male Lewis rats were divided into 4 groups of 5 rats each and gavaged either with 2 ml of water (control) or different concentrations of HC (10 mg/ml, 100 mg/ml and 500 mg/ml). In the second experiment in a similar design, lower doses of HC were used (water, 0.1 mg/ml, 0.50 mg/ml and 5.0 mg/ml). The rats were killed after 1 h and three 3 x 3 mm pieces of gastric tissue were removed from each rat and incubated for the determination of prostaglandin E2 and 6-keto-prostaglandin F1 alpha accumulation in the medium measured by radioimmunoassay. At low doses HC inhibits rat gastric mucosal prostaglandin synthesis whereas at higher doses HC stimulates it. This biphasic effect of HC on gastric mucosal prostaglandin synthesis may help explain its role in ulcerogenesis.


The effects of peripheral D-amphetamine, 4-OH amphetamine, and epinephrine on maintained discharge in the locus coeruleus with reference to the modulation of learning and memory by these substances.
D-Amphetamine, 4-OH amphetamine, and epinephrine have been shown in many behavioral studies to facilitate memory when given post-training. The effect of these drugs on the maintained discharge of cells in the locus coeruleus (LC) was investigated using a route of administration (intraperitoneal) and a log-dose range of these drugs comparable to those used in the behavioral experiments. D-Amphetamine profoundly suppressed maintained discharge: an inhibitory effect was observed at every dose (0.1, 1.0, 10.0 mg/kg). In contrast, only the highest dose of 4-OH amphetamine (8.2 mg/kg) inhibited activity in the LC, and this effect was a modest one. Unlike the amphetamines, epinephrine (500 micrograms/kg) elevated maintained discharge. These results are discussed in the context of the hypothesized involvement of the LC in the enhancement of memory by these drugs.


A case of cutaneous polyarteritis nodosa manifested by spiking high fever, arthralgia and macular eruption like adult-onset Still's disease.
A 24-year-old Japanese woman was admitted for investigation of recurrent spiking high fever associated with a macular eruption of the upper extremities associated with fever and polyarthralgia. These symptoms were self-limiting but recurrent and seemed to be consistent with a diagnosis of adult-onset Still's disease (AOSD). However, livedo reticularis was detected on the lower extremities, suggesting the presence of vasculitic disease rather than AOSD. Investigation did not reveal any evidence of visceral involvement. Skin biopsy of the affected lower extremity demonstrated a necrotizing vasculitis of small muscular artery and confirmed a diagnosis of cutaneous polyarteritis nodosa (PAN) rather than AOSD. Treatment with 30 mg of prednisolone daily improved the skin lesions and the recurrent spiking high fever and the arthralgia were resolved. PAN and AOSD are clinically similar, and discrimination may be sometimes difficult. The presence of livedo reticularis and the finding of a characteristic skin biopsy appearance may be diagnostically useful to distinguish PAN from AOSD. Indeed, the clinical features of cutaneous PAN may be more similar to AOSD than systemic PAN, and a skin biopsy may be necessary to distinguish cutaneous PAN from AOSD in some cases.


Formulation and stability of iopentol.
The autoclaving studies which were performed to develop a formulation of iopentol are presented. The same formulation as used for iohexol was found to be suitable for iopentol. It was shown that it is essential to use a buffer such as trometamol to lower the pH during autoclaving in order to achieve a low level of deiodination after autoclaving. Results from the on-going stability studies indicate a good stability of iopentol finished product.


Progress in the research of Radix Astragali in treating chronic heart failure: effective ingredients, dose-effect relationship and adverse reaction.
Radix Astragali, a Chinese herbal medicine possessing important cardiovascular pharmacologic effects, is widely applied for the treatment of chronic heart failure (CHF) in clinical practice. This paper summarizes briefly the researches in the last 10 years on its chemical compositions, effective ingredients for improving cardiac function, dose-effect relationship in treating CHF, and adverse reactions that occurred in clinical practice.


Pharmacologic effect of recombinant human IFN-alpha, continuously released from a matrix prepared from a polyglycerol ester of fatty acids, on 2',5'-oligoadenylate synthetase activity in murine liver.
The objective of this study was to assess the pharmacologic effect of continuously released recombinant human interferon-alpha (rHuIFN-alpha) in the liver, the target organ of chronic hepatitis B and C, using 2',5'-oligoadenylate synthetase (2',5'-OAS) activity as an indicator of an antiviral state. A cylindrical matrix prepared from tetraglycerol dipalmitate (TGDP), a polyglycerol ester of fatty acids (PGEF), released rHuIFN-alpha in a pseudo-zero-order manner for about 1 week after implantation into mice, without any major loss of rHuIFN-alpha biologic activity during the release period. To evaluate the pharmacologic effect of the rHuIFN-alpha continuously released from this type of matrix, we established a murine test system. Bolus injections of rHuIFN-alpha solution at three doses increased 2',5'-OAS activities in murine liver extract and serum in a dose-dependent manner, indicating that this system is suitable for evaluating rHuIFN-alpha activity. After subcutaneous insertion of TGDP-matrix implants containing 5.5x10(7) IU rHuIFN-alpha per animal, 2',5'-OAS activities in both liver extracts and serum increased rapidly and remained high for over 1 week. Subcutaneous injections of an equivalent total dose (5.0x10(7) IU/animal per week) of rHuIFN-alpha solution in three or seven fractions prolonged 2',5'-OAS activities compared with a single bolus injection. Comparing 2',5'-OAS activity on day 7 and the portion of the area under the 2',5'-OAS activity-time curve above the normal level (deltaAUC) between the TGDP-matrix implant and multiple injections of the solution revealed that continuously released rHuIFN-alpha has an effect almost equivalent to that of three or seven injections of the solution per week.


[Management of hypertrophic cardiomyopathy - the most common inherited heart disease].
Hypertrophic cardiomyopathy is the most common cardiogenetic disease affecting 1/500-1/1 000 individuals. Dyspnea is common but chest pain, dizziness or fainting may also cause considerable limitation for the patient. The diagnosis can be suspected from ECG. Echocardiography confirms hypertrophy of at least 15 mm, usually in the septum. If the obstruction of the outflow tract is severe, myectomy or alcohol ablation can relieve symptoms. Genetic evaluation of family members is advisable. To reduce symptoms, betablockers are used; verapamil or disopyramide are alternatives. Atrial fibrillation is often prevalent and requires special attention concerning anticoagulation and rhythm or rate control. An end-stage heart failure warrants advanced treatment options such as cardiac resynchronization therapy, ventricular assist devices or heart transplant. Sudden cardiac death is unpredictable and evaluation of risk markers is important to identify potential candidates for an implantable defibrillator.


Paediatric liver transplantation [corrected] in Melbourne: the first 50 patients.
To report the outcomes of the first 50 paediatric patients who have undergone liver transplantation (LT) at the Victorian Liver Transplant Unit, a retrospective review of case records was carried out. From December 1988 to December 2000, 108 patients 18 years or younger were referred for LT; 50 of these underwent a total of 53 transplants. The most common indications were biliary atresia (32%), metabolic disease (26%), and acute hepatic necrosis (26%). The majority of deaths (6/7) occurred in the 1st week after LT. The actuarial survival at 1 year was 88% (95% CI 75% to 94%) and at 10 years 85% (95% CI 71% to 93%). Survival rates were highest for children aged 3 to 14 years (95%) and lowest in those weighing less than 8 kg at the time of LT (66%). All 43 survivors are attending age-appropriate activities including kindergarten, school, and employment. The survival of patients undergoing LT in this unit compares favourably with those recorded by the Australia and New Zealand Transplant Registry and is commensurate with that reported by larger paediatric transplant programs overseas.


Gas Chromatographic Detection of Pre-Processing Spoilage of Bacterial Origin in Pet Foods.
Gas chromatographic analysis of sterile canned pet foods revealed the presence of metabolites indicative of pre-processing spoilage. The number and type of aerobic and anaerobic bacteria in each raw ingredient were determined. It was found that pre-processing spoilage was due to the action of the predominant anaerobe or anaerobes in the major ingredient by comparing the gas chromatographic profiles of the organisms with the chromatograms of the finished products. This type of spoilage was more common in pet foods manufactured by cereal companies who may use dead stock than in similar products manufactured by meat packing companies.


Hypoglycaemic activity of alpha, alpha trehalose-6-phosphate.
Alpha-alpha-trehalose-6-phosphate, synthesized by Oke was screened for hypoglycaemic activity. Alloxan-induced diabetic albino rats and fasted-rabbits were used in the study. The inhibitory activity of trehalose-6-phosphate on trehalase was also assayed. The study shows that alpha-alpha-trehalose-6-phosphate is a glucose analogue with potent anti-hyperglcaemic activity as shown by its hypoglycaemic response in fasted rabbits. The ability of alpha-alpha-trehalose-6-phosphate to attenuate the diabetic toxicity in alloxan-induced diabetic rats confirmed its potent anti-diabetic activity. The mechanism of action of this synthesized compound may be linked with its ability to inhibit trehalase, and increase the activity of the superoxidase dimutase present in the beta-cells of the alloxan-diabetic rats and also being a glucose analogue according to Puls principle, alpha-alpha-trehalose-6-phosphate is able to influence the intermediate metabolism of carbohydrate.


The Common Fund Initiative and Its Implication for Advancing Exercise and Physical Activity Research in Child Health.
NIH Director Francis Collins noted that the Common Fund initiative would lead to unprecedented insights into the mechanisms responsible for the health effects of physical activity. He noted: “Armed with this knowledge, researchers and clinicians may one day be able to define optimal physical activity recommendations for people at various stages of life, as well as develop precisely targeted regimens for individuals with particular health needs.” Given the ominous burden of physical inactivity-related diseases and conditions in otherwise healthy children, and the growing number of children who survive chronic diseases in whom we know little about what constitutes healthy exercise, it is essential that the community of child health researchers develop compelling strategies and proposals in response to the unique opportunity offered through the Common Fund mechanism.


Immunosuppressive medications, clinical and metabolic parameters in new-onset diabetes mellitus after kidney transplantation.
New-onset diabetes after transplantation (NODAT) is a growing concern in transplantation. All modifiable risk factors are not yet identified. We assessed the relationship between baseline clinical and biochemical parameters and NODAT. Eight-hundred and fifty-seven in-Caucasian renal transplant recipients were included. Charts were individually reviewed. The follow-up was 5.3 years (ranges: 0.25-20.8; 5613 patient-years). The incidence of NODAT was 15.0%, 18.4% and 22.0% at 10, 15 and 20 years following transplantation. Age, body mass index (BMI), glucose (all P < 0.0001) and triglycerides [hazard ratio (HR) per 1 mmol/l: 1.44 [1.17-1.77], P = 0.0006] were potent risk factors whereas steroid withdrawal (HR: 0.69 [0.47-1.01], P = 0.0601) reduced the risk. As compared to cyclosporine, sirolimus (HR: 3.26 [1.63-6.49], P = 0.0008) and tacrolimus (HR: 3.04 [2.02-4.59], P < 0.0001) were risk factors for NODAT. The risk of NODAT was comparable for sirolimus (HR: 2.35 [1.06-5.19], P = 0.0350) and tacrolimus (HR: 2.34 [1.46-3.75], P = 0.0004) after adjustments on age, BMI, glucose and steroid withdrawal; however, unlike sirolimus, tacrolimus remained significant after adjustment on triglycerides. The risk of NODAT appeared similar, but its pathophysiology seemed different in sirolimus- and tacrolimus-treated patients; this observation needs confirmation. However, main independent risk factors were age, BMI, initial glucose and triglycerides.


Caffeine enhances acetylcholine release in the hippocampus in vivo by a selective interaction with adenosine A1 receptors.
Caffeine is a commonly used drug that increases arousal, a condition associated with increased cholinergic activity in the mammalian cerebral cortex including the hippocampus. We have used the technique of microdialysis in association with microbore high-performance liquid chromatography to investigate the effects of caffeine on the extracellular levels of acetylcholine in the hippocampus of awake, freely moving rats. The oral administration of caffeine dose-dependently (3-30 mg/kg) increased the extracellular levels of acetylcholine. This increase was completely blocked when the microdialysis probe was perfused with the Na(+)-channel blocker tetrodotoxin, and strongly attenuated when a Ca(2+)-free Ringer solution was used. The effect of caffeine on hippocampal acetylcholine release was concentration-dependently counteracted by local perfusion of an A1 receptor agonist, N6-cyclopentyladenosine (0.1-1 mumol/liter), but not by the A2 receptor agonist, CGS 21680 (10 mumol/liter). Neither agonist affected base-line acetylcholine release at these concentrations. These results demonstrate that acetylcholine release in the hippocampus is under tonic inhibitory control of the endogenous neuromodulator adenosine, and that orally administered caffeine enhances action potential-dependent vesicular acetylcholine release by antagonism of local A1 receptors. Hence, the data provide a possible link between adenosine A1 receptors in the hippocampus, increased cholinergic activity and the psychostimulant effects of caffeine.


Identification of key genes and pathways associated with osteogenic differentiation of adipose stem cells.
Adipose stem cells (ASCs) are considered a great alternative source of mesenchymal stem cells (MSCs) and have shown great promise on tissue engineering and regenerative medicine applications, including bone repair. However, the underlying mechanisms regulating the osteogenic differentiation of ASCs remain poorly known. Gene expression profiles of GSE63754 and GSE37329 were downloaded from gene expression omnibus database. R software and Bioconductor packages were used to compare and identify the differentially expressed genes (DEGs) before and after ASC osteogenic differentiation. The common significant DEGs between GSE63754 and GSE37329 were then subjected to gene ontology (GO) enrichment analysis, ingenuity pathway analysis (IPA), and protein-protein interactions (PPIs) networks analysis. One of the central node genes FOXO1 was selected for further investigation. A total of 142 up- and 69 downregulated genes were aberrantly expressed in both GSE63754 and GSE37329. GO analysis revealed that these DEGs were associated with extracellular matrix organization, proteinaceous extracellular matrix, and Wnt-protein binding. IPA analysis showed that canonical pathways, such as FXR/RXR activation, adipogenesis pathway, and LXR/RXR activation, were involved in regulating osteogenic differentiation of ASCs. A total of three subnetworks and 39 nodes were identified with PPI network and MCODE plugin. Moreover, suppression of one central node gene FOXO1 inhibited the osteogenic differentiation of ASCs. Our study provides a registry of genes and pathways that play important roles in regulating osteogenic differentiation of ASCs, which might have potential therapeutic applications in bone regeneration and bone tissue engineering.


Fragile but real: children's capacity to use newly acquired words to convey preverbal memories.
There is ongoing debate about children's ability to use subsequently acquired language to describe preverbal experiences. This issue was addressed experimentally in this investigation using a novel paradigm. Two-year-old children who lacked color words were individually taught to activate a bubble machine by selecting a particular color of bubble solution. The children then participated in weekly, experimenter-provided activities that fostered their acquisition of the color labels. After 2 months, their ability to apply the newly acquired words in reporting the original event was assessed. A significant proportion of the children demonstrated verbal recall when prompted in the presence of physical reminders of the event. These findings indicate that some early, preverbal memories are translated into words at a later time.


[Effect of endogenous carbon monoxide on oxidant-mediated multiple organ injury following limb ischemia-reperfusion in rats].
To determine the role of endogenous carbon monoxide(CO) in oxidant-mediated organ injury following limb ischemia-reperfusion (I/R) in rats. Sixty-four SD rats were divided into 4 groups: Sham group, Sham + zinc protoporphyrin (ZnPP, an inhibitor of heme oxygenase activity), 2-hour ischemia followed by 4-hour reperfusion (I/R) group and I/R + ZnPP group. Carboxyhemoglobin (COHb) level in the artery blood, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the lung, heart, liver and kidney were detected. The 24-hour survival rate of rats was studied. Compared with the sham group, the COHb level and MDA content significantly increased, while the SOD activity and the survival rate significantly decreased in I/R group (P < 0.05). Compared with the I/R group, MDA content significantly increased, while the SOD activity, the 24-hour survival rate and COHb level significantly decreased in I/R + ZnPP group (P < 0.05, respectively). Limb I/R could lead to the oxidant-mediated multiple organ injury accompanied by the increase of CO level which play an important role in the defense against I/R-induced remote multiple organ injury in rats.


Dietary eritadenine-induced alteration of molecular species composition of phospholipids in rats.
The effect of dietary eritadenine, a hypocholesterolemic compound found in the mushroom Lentinus edodes, on the fatty acid and molecular species profiles of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in the plasma and liver microsomes was investigated in relation to the hypocholesterolemic action of the compound in rats. Animals were fed the control or eritadenine-supplemented (50 mg/kg diet) diet for 14 d. Eritadenine supplementation significantly decreased the plasma concentrations of cholesterol and phospholipids, but not triglycerides. The PC/PE ratio of liver microsomes, but not plasma, was also markedly decreased by eritadenine. Eritadenine supplementation was found to increase the proportion of 18:2n-6 and, inversely, to decrease the proportion of 20:4n-6 and 22:5n-6 in plasma PC and liver microsomal PC and PE, indicating that eritadenine depressed the metabolism of linoleic acid. The effect of eritadenine on the profile of n-3 fatty acids was dissimilar in PC and PE. These changes in fatty acid composition were selectively reflected in the molecular species composition of both PC and PE; the extent of increase in 16:0-18:2 molecular species or decrease in 18:0-20:4 molecular species was apparently greater than that of other molecular species containing 18:2 or 20:4 in the sn-2 position. These results suggest that, in addition to the decrease in liver microsomal PC/PE ratio, the alteration of plasma PC molecular species composition might also participate in the hypocholesterolemic action of eritadenine.


Cyclic Steroid Glycosides from the Starfish Echinaster luzonicus: Structures and Immunomodulatory Activities.
Five new steroid glycosides, luzonicosides B-E (2-5), belonging to a rare structure group of marine glycosides, containing carbohydrate moieties incorporated into a macrocycle, and a related open carbohydrate chain steroid glycoside, luzonicoside F (6), were isolated from the starfish Echinaster luzonicus along with the previously known cyclic steroid glycoside luzonicoside A (1). The structures of compounds 2-6 were established by extensive NMR and ESIMS techniques as well as chemical transformations. Luzonicoside A (1) at concentrations of 0.01-0.1 μM was shown to be potent in lysosomal activity stimulation, intracellular ROS level elevation, and NO synthesis up-regulation in RAW 264.7 murine macrophages. Luzonicoside D (4) was less active in these biotests.


Impact of valve surgery on serum osteopontin levels in patients with mitral regurgitation.
Osteopontin (OPN), a sialoprotein present within atherosclerotic lesions, especially in calcified plaques, is linked to the progression of coronary artery disease and heart failure. We assessed the impact of valve surgery on serum OPN and left ventricular (LV) function in patients with mitral regurgitation (MR). Thirty-two patients with severe MR scheduled for surgery were included in the study. Echocardiography markers were assessed preoperatively and at 3 months following the surgery and matched with the serum OPN levels. Valve surgery was associated with a reduction of the ejection fraction (EF) from 55.2 ± 6.3 to 48.8 ± 7.1% after surgery, p < 0.001. Following surgery, the OPN level was significantly higher than preoperatively (mean 245, range 36-2,284 ng/ml vs. 76, 6-486 ng/ml, p = 0.007). Preoperative OPN exhibited a slight negative correlation with the EF (r = -0.35, p = 0.04), and a moderate correlation with vena contracta (r = -0.38, p = 0.02). There were no other meaningful correlations between conventional echocardiographic parameters and OPN. Following valve surgery due to severe MR, patients exhibited a decrease in EF and an increase in OPN levels. The assessment of preoperative OPN failed to strongly predict probable LV dysfunction.


Ganciclovir. A review of its antiviral activity, pharmacokinetic properties and therapeutic efficacy in cytomegalovirus infections.
Ganciclovir is a nucleoside analogue with antiviral activity in vitro against members of the herpes group and some other DNA viruses. It has demonstrated efficacy against human cytomegalovirus infections and should be considered a first-line therapy in the treatment of life- or sight-threatening cytomegalovirus infection in immunocompromised patients. Clinical efficacy varies with the underlying aetiology of immunocompromise and the site of disease, and prompt diagnosis and early treatment initiation appear to improve the response. In patients with cytomegalovirus pneumonia, particularly bone marrow transplant recipients, concomitant administration of cytomegalovirus immune globulin may significantly improve clinical outcome. Maintenance therapy to prevent recurrence is usually required by bone marrow transplant recipients until the recovery of adequate immune function, whereas AIDS patients may require indefinite ganciclovir maintenance therapy to prevent disease progression, as ganciclovir (like other antivirals) does not eradicate latent viral infection. Haematological effects occur relatively frequently during ganciclovir administration but are usually reversible. Ganciclovir has not been directly compared with other antiviral drugs because of the absence until recently of other effective treatments. However, comparative studies with foscarnet, particularly in cytomegalovirus retinitis, will be of considerable interest. Thus, ganciclovir represents a major advance in the therapy of severe cytomegalovirus infections in immunocompromised patients. Comparative studies, and investigation of ways of reducing toxicity (intravitreal administration; concomitant use of stimulants of haematopoiesis; use in conjunction with other antivirals with differing mechanisms of action), may further expand its eventual role.


An unusual neurological feature of HIV-1 encephalopathy: Gerstmann's syndrome.
The authors describe the first case in literature of Gerstmann's syndrome (agraphia, acalculia, finger agnosia) occurred in HIV correlated encephalopathy developed as the first severe manifestation of HIV infection in a patient with prevalent white matter neuroradiologic alterations. The PDL rapidly extended from the left subcortical parietal-occipital regions to the pre-rolandic one, with subsequent involvement of the corpus calosum splenium and the bilateral temporal lobes white matter. The authors indicate the extent of the lesions and the involvement of the interhemispheric connection fibres as the pathogenetic mechanism of the "Gerstmann syndrome", that until today has not been reported in the literature of the wide variety of AIDS dementia complex. The administration of 1 g of zidovudine for about 9 months did not avoid the establishing of the neurologic damage, but the sudden suspension of the drug could have enhanced the exacerbation of inflammation and the involvement of areas whose lesion is classically believed responsible for cognitive impairment.


Human immunodeficiency virus in breast milk.
As the acquired immunodeficiency syndrome (AIDS) epidemic spreads to the pediatric population, a recommendation is made for more research on mother-to-infant human immunodeficiency virus (HIV) transmission, in light of current policies, and the scientific community is challenged to re-evaluate its attitude to the pathogenesis of HIV transmission by breast milk.


Isolated intraosseous lipoma of the ethmoid sinus.
Intraosseous lipoma is an extremely rare variant of lipoma. It is most commonly found in the appendicular skeleton and only very rarely in skull base. We present a 38-year-old male patient who was admitted to our clinic with the symptoms of nasal obstruction and headache. Computed tomography (CT) showed a hyperdense area with surrounding calcification was demonstrated in the anteromedial part of ethmoid sinus. Endoscopic left anterior ethmoidectomy was performed and the mass was totally excised. Histopathological examination was consistent with Milgram's stage 1 intraosseous lipoma. To our knowledge, this is the first reported case of isolated intraosseous lipoma of the ethmoid sinuses in the literature.


Proteins immobilization on the surface of modified plant viral particles coated with hydrophobic polycations.
Two hydrophobic cations based on poly-N-ethyl-vinylpyridine were used to produce biologically active complexes. The complexes obtained from tobacco mosaic virus (TMV) spherical particles (SPs), hydrophobic polycation, and a model protein were stable and did not aggregate in solution, particularly at high ionic strengths. The nucleic acid-free SPs were generated by thermal remodeling of the TMV (helical rod-shaped plant virus). The model protein preserved its antigenic activity in the ternary complex (SP-polycation-protein). Immobilization of proteins on the surface of SPs coated with hydrophobic cation is a promising approach to designing biologically active complexes used in bionanotechnologies.


Does the presence of an upper extremity injury affect outcomes after major trauma?
There is a paucity of information about the impact of upper extremity (UE) injuries on patient outcomes, particularly after major trauma. Data were obtained from a statewide trauma registry. Cases were defined as major trauma cases (Injury Severity Score > 15) with (UE group) and without (no-UE group) an associated upper extremity injury. Multivariate analysis was performed to identify independent predictors of outcome. Major trauma patients with UE injury were 1.5 times (p = 0.011) more likely than the no-UE group to have a length of stay greater than 7 days. After adjusting for age, mechanism of injury, and Injury Severity Score, UE injury was not an independent predictor of discharge destination. In major trauma patients, the presence of an upper extremity injury is a significant predictor of length of stay, indicating a greater complexity and cost of care associated with this group of major trauma patients.


Changes in the serum lipid profile in man during 24 months of arctic residence.
The influence of the severe climate and geographical conditions at the Svalbard archipelago (78-79 degrees N) on serum lipid levels were measured in Caucasian miners who had arrived from the southern part of Ukraine and Russia (48 degrees N). The persons included in the study were randomly divided in five groups according to their time of living (1, 3, 6, 12 and 24 months) at Svalbard. Blood sampling took place during a two week period in January, when the Svalbard archipelago is into its polar night. General elevated levels of triglycerides were found in group I-III (1, 3 and 6 months stay), whereas the values measured in group IV and V (12 and 24 months stay) were somewhat lower. This apparent decline in triglycerides was paralelled by generally elevated levels of HDL cholesterol. The serum level of phospholipids was similar in all groups. All the level of free fatty acids was apparently higher in groups IV and V, particularly 18:3 and 16:1. These results may be indicative of a rise in triglyceride consumption after about a 12 month stay in the archipelago. Besides, the elevated levels of 18:3 and 16:1 fatty acids imply dietary modifications of the serum fatty acids.


The development of a questionnaire to measure the severity of symptoms and the quality of life before and after surgery for stress incontinence.
To develop and validate a short questionnaire recording the severity of symptoms and the quality of life, pre- and post-operatively, in women undergoing surgery for stress incontinence. The questionnaire was designed from previously designed questionnaires and pilot studies. In this study, it was tested for validity and reliability using standard psychometric techniques. Data from the questionnaire are reported to a national database, which generates reports for comparison of surgical procedures, departments and surgeons. Tertiary referral urogynaecology units at Baerum and Aker Hospitals, Norway, and 22 Norwegian departments of gynaecology. Fifty and 65 women with stress incontinence from Baerum and Aker hospitals, respectively, and 628 women with stress incontinence from 22 Norwegian departments of gynaecology. Stress Incontinence, Urge Incontinence and Quality of Life Indices were constructed. Internal consistency of the indices was measured by Cronbach's alpha and test-retest reliability by Bland-Altman plots. Stress Incontinence Index was tested against stress test and Urge Incontinence Index and Stress Incontinence Index were tested against the 24-hour pad test. Quality of Life Index was tested against a part of the King's College Hospital Quality of Life Questionnaire. Face and content validity. Missing values of the questionnaire. Reliability and internal consistency. Criterion validity. The 628 women completed a mean of 98.2% of all the questions. The content validity was good. The reliability was good in terms of test-retest reliability and internal consistency. The criterion validity of stress and urge incontinence was tested against the stress and 24 hour pad tests. The correlation between the indices and the objective tests was moderate. The correlation between the Quality of Life Index and the physical and social part of the previously validated King's College Hospital questionnaire was good. The questionnaire is easy to understand and complete and is a valid and reliable instrument for assessment of the symptoms of incontinence and the quality of life. Pre- and post-operative evaluation of women with stress incontinence was performed in a standardised setting in 22 of the 37 Norwegian departments and the reporting to a national database was successful. Comparison of surgical procedures, departments and surgeons is possible.


A cascade of care for diabetes in the United States: visualizing the gaps.
A "cascade-of-care" concept helped to address implementation gaps in HIV care. To develop a similar cascade for U.S. diabetes care to visualize gaps in awareness of diagnosis, engagement, and treatment. Nationally representative cross-sectional surveys. 2007 to 2012 NHANES (National Health and Nutrition Examination Surveys). Nonpregnant civilians aged 18 years or older. Standardized data collection and laboratory procedures. Diabetes care was benchmarked against recent diabetes and cardiovascular risk reduction guidelines. In 2012, an estimated 28.4 million (11.8%) U.S. adults had diabetes, of whom 20.5 million (72.2%) were aware of their diagnosis. Among diagnosed adults, 95.3% had a usual care provider and 91.7% made 2 or more visits in the past year. In contrast, among undiagnosed adults, 84.5% had a usual care provider and 66.5% reported 2 or more visits in the past year. Among diagnosed adults, 63.7%, 65.5%, 56.6%, and 80.6% met individualized hemoglobin A1c, blood pressure (BP <140/80 mmHg), lipid (low-density lipoprotein [LDL] cholesterol <2.6 mmol/L [<100 mg/dL]), and nonsmoking goals, respectively; 26.7% met combined ABC (hemoglobin A1c, BP, and LDL cholesterol) targets, and 21.3% met combined ABC targets and did not smoke. Among undiagnosed adults, 77.0%, 57.9%, 36.0%, and 77.9% met hemoglobin A1c, BP, lipid, and nonsmoking goals, respectively; 22.1% met combined ABC targets; and 18.8% met combined ABC targets and were nonsmokers. Institutionalized and noncivilian persons are not surveyed in NHANES. Three of 10 adults with diabetes remain undiagnosed, which may be related to less access to care. Compared with diagnosed adults, undiagnosed adults have less elevated hemoglobin A1c levels, less lipid treatment and worse control, and similarly poor BP and combined ABC control regardless of smoking status. Addressing these care gaps in both groups would prevent long-term complications. None.


Resource competition and shared natural enemies in experimental insect communities.
Much theory has been developed to explore how competition for shared resources (exploitation competition) or the presence of shared natural enemies (apparent competition) might structure insect and other communities. It is harder to predict what happens when both processes operate simultaneously. We describe an experiment that attempted to explore how shared natural enemies and resource competition structured a simple experimental insect community. Replicated communities were assembled in population cages consisting of the aphid species Acyrthosiphon pisum and Megoura viciae either alone or competing for a resource, their shared host plant Vicia faba. Each combination was set up with and without the parasitoid Praon dorsale which attacked both species of aphid. Population dynamic data show that interspecific resource competition was the dominant process structuring the community. Though juvenile parasitoids could develop successfully inside hosts of both species, they failed to suppress either aphid below their carrying capacities and were unable to persist on one species. We suggest that intense resource competition may reduce the value of individual aphids as hosts for parasitoids such that their population growth rate is less than zero and discuss whether this phenomenon occurs in natural and agricultural communities.


Substituting Sedentary Time With Light and Moderate to Vigorous Physical Activity is Associated With Better Cardiometabolic Health.
The purpose of the current study was to apply a more novel approach to systematically examine (1) associations of clustered cardiometabolic risk and cardiometabolic risk factors and (2) theoretical substitution of sedentary time with either sleep, light physical activity (LPA), or moderate to vigorous physical activity (MVPA) and substituting LPA with MVPA. Physical activity and sleep were objectively measured in 410 Flemish adults [55.5 (9.6) y, 64% men] with a SenseWear Pro 3 Armband. Cardiometabolic risk factors (obesity, hyperglycemia, dyslipidemia, and hypertension) and cardiorespiratory fitness were objectively measured. Isotemporal substitution analyses were performed to assess the associations between substituting time from a potentially negative behavior into another potentially positive behavior. Theoretical substitution of sedentary time with MVPA was associated with decreased clustered cardiometabolic risk, b = -0.06 (-0.08 to -0.04), and substituting LPA with MVPA was associated with a decrease in clustered cardiometabolic risk, b = -0.08 (-0.11 to -0.04). Substituting sedentary time with LPA or sleep improved high-density lipoprotein-cholesterol, systolic and diastolic blood pressures, and waist circumference. Theoretical replacement of sedentary time with either sleep, LPA, or MVPA was positively associated with improved cardiometabolic risk factor status. Interventions for increasing cardiometabolic health can focus on replacing sedentary time with either sleep, LPA, or MVPA depending on the risk parameters that need to be targeted.


[Detection of respiratory viruses in children with acute lower respiratory tract infection: an analysis of 5,150 children].
To investigate the distribution of respiratory viruses on throat swabs in hospitalized children with acute lower respiratory tract infection (ALRTI). A total of 5,150 children with ALRTI who were admitted to Hebei Children's Hospital between March 2014 and February 2015 were enrolled to investigate the distribution of respiratory viruses in children with ALRTI. Direct immunofluorescence assay was performed for throat swabs from these children to detect influenza virus A (FA), influenza virus B (FB), adenovirus (ADV), respiratory syncytial virus (RSV), and parainfluenza virus types 1, 2, and 3 (PIV-1, PIV-2, and PIV-3). Of all the 5,150 throat swabs from hospitalized children, 2,155 (41.84%) had positive virus detection results. RSV had the highest detection rate (1,338 cases/25.98%), followed by PIV-3 (439 cases/8.52%) and FA (166 cases/3.22%), and 29 patients had mixed infection with 2 viruses. With the increasing age, the detection rates of viruses tended to decrease (χ2=279.623; P<0.01). The positive rate of RSV increased gradually from September, and reached the peak value (60.09%) in November; the lowest positive rate occurred in June (1.51%). The positive rate of PIV-3 was the highest in May (21.38%) and the lowest in November (1.77%). The distribution of viruses in children with ALRTI varies with age and season, with RSV prevalence in autumn and winter and PIV-3 prevalence in spring and summer. RSV is the most common viral pathogen that causes ALRTI in hospitalized children.


[Clinical aspects and diagnosis of cerebral hemorrhage].
Primary intracerebral hemorrhages are often dramatic and lethal. Therapeutic possibilities are limited. Multivariate studies show that patient age, hematoma volume, ventricular extension of the hemorrhage, and level of consciousness are the decisive prognostic parameters. This overview shows that the etiological spectrum is larger than suggested by the classic paradigm of "either hypertensive or cryptic angioma". The variability of clinical presentation can be put into a systematic order by means of clinical neurological examination and imaging techniques. Experiences with 62 consecutive cases from the Klosterneuburg Stroke Data-Bank are reported.


[Observation of penetration, distribution and accumulation in human renal proximal tubular epithelial cells by aristololactam-I].
To study whether aristololactam I (AL-I) can enter renal proximal tubular epithelial cells and the situation of intracellular distribution and accumulation. Cultured human renal proximal tubular epithelial cell line (HK-2) was used as the subject. Intracellular fluorescence from AL-I and its distribution are examined by fluorescence microscopy after a treatment with different concentration of AL-I, the intracellular accumulation of AL-I was also investigated by incubated cells in AL-I -free medium for 48 h after washing-out the media containing AL-I. After treatment of AL-I (concentration from 5 microg x mL(-1) to 20 microg x mL(-1)), glaucous fluorescence could be observed inside renal proximal tubular epithelial cells at 0.5 h, and the fluorescence distributed only in cytoplasm while not be observed in nuclei. Moreover, the fluorescence of AL-I could be kept in cytoplasm for more than 48 h after washing out the media containing AL-I . AL-I is able to enter renal proximal tubular epithelial cells in short time and accumulate in cytoplasm, but not enter nuclei. This property may contribute to the cytotoxic mechanism of renal injury induced by AL-I, which may partially explain the persistent renal toxicity of AAs and its metabolites in the development of aristolochic acid nephropathy.


Evaluation of DNA recombinant methodologies for the diagnosis of Plasmodium falciparum and their comparison with the microscopy assay.
Since 1984, DNA tests based on the highly repeated subtelomeric sequences of Plasmodium falciparum (rep 20) have been frequently used in malaria diagnosis. Rep 20 is very specific for this parasite, and is made of 21 bp units, organized in repeated blocks with direct and inverted orientation. Based in this particular organization, we selected a unique consensus oligonucleotide (pf-21) to drive a PCR reaction coupled to hybridization to non-radioactive labeled probes. The pf-21 unique oligo PCR (pf-21-I) assay produced DNA amplification fingerprints when was applied on purified P. falciparum DNA samples (Brazil and Colombia), as well as in patient's blood samples from a large area of Venezuela. The performance of the Pf-21-I assay was compared against Giemsa stained thick blood smears from samples collected at a malaria endemic area of the Bolivar State, Venezuela, at the field station of Malariología in Tumeremo. Coupled to non-radioactive hybridization the pf-21-I performed better than the traditional microscopic method with a r = 1.7:1. In the case of mixed infections the r value of P. falciparum detection increased to 2.5:1. The increased diagnostic sensitivity of the test produced with this homologous oligonucleotide could provide an alternative to the epidemiological diagnosis of P. falciparum being currently used in Venezuela endemic areas, where low parasitemia levels and asymptomatic malaria are frequent. In addition, the DNA fingerprint could be tested in molecular population studies.


Involvement of the thymus and cellular immune system in craniofacial malformation syndromes.
Craniofacial structures, the aortic arch, thymus, and parathyroid glands all arise from the embryologic pharyngeal pouches, and DiGeorge and Job craniofacial malformation syndromes have defined immunologic deficiencies. The question addressed by this study is whether patients with other pharyngeal pouch malformations could also have immunologic abnormalities. Twelve patients, 4 female and 8 male, were selected at random from the Tampa Bay Craniofacial Center. Their diagnoses included: cleft lip/cleft palate, hemifacial microsomia/Goldenhar syndrome, Treacher-Collins syndrome, craniofacial hemangiomata, cranio-synostosis syndromes, and Tessier 13 cleft. Fresh blood samples were analyzed against age-matched controls for immunoglobin number, using immunoelectrophoresis, T-cell, B-cell, and natural killer cell quantity via Coulter counter and monoclonal antibody labeling, as well as lymphocyte stimulation and response functions with phytohemagglutinin, concanavalin A, pokeweed mitogen, and Staphylococcus aureus mitogens. All patients studied had some abnormality of their immune systems. Seven had specific T-cell abnormalities and three patients had abnormalities in all three categories studied. This indicates that patients with any pharyngeal pouch malformation may have an abnormality of the immune system.


[Clinical experience with cefoxitin in bacterial infections of children (author's transl)].
Using a new cephamycin antibiotic, cefoxitin, clinical studies were carried out, and the following results were obtained: (1) Cefoxitin was administered to 52 children with 38 respiratory tract infections, 2 urinary tract infections, 1 acute otitis media and others, who ranged in age from 2 months to 11 years old. (2) Cefoxitin was given intravenously at a daily dose of 27.3 mg/kg to 110.5 mg/kg. (3) The overall efficacy rate was 80% in 46 cases, i.e., excellent in 10, good in 27, fair in 7 and poor in 2. (4) Clinical side effects were not seen except exanthema in 1 patient. Elevation of GOT and GPT was seen in 2 patients, elevation of A1-P in 1 and eosinophilia in 2 by laboratory tests. However, these findings were alleviated rapidly following the cessation of therapy.


Multiple Gastritis Cystica Profunda in a Patient without Gastric Surgery History.
Gastritis cystica profunda (GCP) is a rare condition characterized by polypoid cystic ectasia of the submucosal layer with cystic dilatation of the glandular structures of the stomach. The probable pathogenesis refers to degradation of the integrity of the muscularis mucosa and migration of epithelial cells to the submucosal layer. The most common cause known in the literature for this occurrence is previous stomach surgery. A case presented with two GCP lesions in different locations in the stomach without any history of previous surgery.


Drug administration errors: a prospective survey from three South African teaching hospitals.
This prospective study was undertaken to determine the incidence of drug administration errors by anaesthetists at three tertiary South African hospitals. Hospitals A and C treat adults predominantly, whereas Hospital B is a paediatric hospital. Anaesthetists completed an anonymous study form for every anaesthetic performed over a six-month period. They were asked to indicate whether or not an error or near-miss had occurred and if so, the details thereof. A total of 30,412 anaesthetics were administered during the study period. The response rate and combined incidence of errors and near-misses was as follows: Hospital A 48.8% (1:320), B 81.3% (1:252) and C 48.1% (1:250). The overall response rate was 53% and the combined incidence was 1:274. Neither the experience of the anaesthetist nor emergency surgery influenced whether an error occurred or not. Most errors occurred during the maintenance phase of anaesthesia. The most common errors were those of substitution. At the paediatric hospital, incorrect dose was as frequent an error as substitution. Of all errors, 36.9% were due to drug ampoule misidentification; of these the majority (64.4%) were due to similar looking ampoules. Another 21.3% were due to syringe identification errors. No major complication attributable to a drug administration error was reported. Despite an increasing awareness of the problem together with suggestions in the literature to reduce the incidence, drug administration errors remain fairly common in South Africa. Failure to institute suggested solutions will continue to compromise patient safety.


Inhibitory action of clozapine on rat ventral tegmental area dopamine neurons following increased levels of endogenous kynurenic acid.
The mode of action by which the atypical antipsychotic drug clozapine exerts its superior efficacy to ameliorate both positive and negative symptoms is still unknown. In the present in vivo electrophysiological study, we investigate the effects of haloperidol (a typical antipsychotic drug) and clozapine on ventral tegmental area (VTA) dopamine (DA) neurons in a situation of hyperdopaminergic activity in order to mimic tentatively a condition similar to that seen in schizophrenia. Increased DA transmission was induced by elevating endogenous levels of the N-methyl-D-aspartate receptor and alpha7(*) nicotinic receptor antagonist kynurenic acid (KYNA; by means of PNU 156561A, 40 mg /kg, i.v.). In control rats, i.v. administered haloperidol (0.05-0.8 mg/kg) or clozapine (1.25-10 mg/kg) was associated with increased firing rate and burst firing activity of VTA DA neurons. However, in rats displaying hyperdopaminergia (induced by elevated levels of KYNA), the effects of clozapine on VTA DA neurons were converted into pure inhibitory responses, including decrease in burst firing activity. In contrast, haloperidol still produced an excitatory action on VTA DA neurons in rats with elevated levels of endogenous brain KYNA. The results of the present study suggest that clozapine facilitates or inhibits VTA DA neurotransmission, depending on brain concentration of KYNA. Such an effect of clozapine may be related to its unique effect in also ameliorating negative symptoms of schizophrenia.


Topical ketanserin attenuates hyperalgesia and inflammation in arthritis in rats.
We investigated effects of topical application of ketanserin, a 5-HT2A receptor antagonist, on hyperalgesia and edema in the arthritic rat, a chronic pain model with inflammation. Unilateral, but not bilateral, arthritis was induced with intra-articular injection of a mixture of kaolin and carrageenan in one side, as indicated by the shortened paw withdrawal latency and an increase in the circumference of the knee joint. Topical application of ketanserin onto skin over the arthritic joint delivered in a mixture of gelatin, glycerol and kaolin produced dose-dependent attenuation of nociceptive and inflammatory effects resulting from intra-articularly injected kaolin/carrageenan. One and 3% ketanserin produced significant or even complete anti-hyperalgesia, as well as a remarkable anti-inflammatory effect (50-70% reduction of edema) while 0.3% ketanserin and placebo failed to produce any effect. Moreover, the effects of ketanserin were maintained for 13 days without decline. In contrast, 3% ketanserin applied to skin of the knee joint on the non-inflamed side for 2 weeks did not alter nociceptive thresholds of the paw and the size of the knee joint in both the inflamed and non-inflamed limbs. These results indicate that 5-HT2A receptors in the periphery play a significant role in the maintenance and/or development of inflammatory pain. The present study suggests that topical ketanserin is a promising direction for potential clinical exploration to relieve established hyperalgesia and inflammation in arthritis without adverse effects and tolerance.


PPAR-gamma dependent and independent effects on macrophage-gene expression in lipid metabolism and inflammation.
Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is highly expressed in lipid-accumulating macrophages of the coronary artery. In light of this, the wide-spread clinical use of thiazolidinediones (TZDs) in the treatment of type II diabetes raises concerns about the role of PPAR-gamma in macrophage function and disease progression. To define the role of PPAR-gamma in macrophage biology, we used homologous recombination to create embryonic stem cells that were homozygous for a null mutation in the PPAR-gamma gene. We demonstrate here that PPAR-gamma is neither essential for nor substantially affects the development of the macrophage lineage both in vitro and in vivo. In contrast, we show it is an important regulator of the scavenger receptor CD36, which has been genetically linked to lipid accumulation in macrophages. Both 15-deoxy-Delta12,14prostaglandin J2 and thiazolidinediones have anti-inflammatory effects that are independent of PPAR-gamma. We show that PPAR-gamma is required for positive effects of its ligands in modulating macrophage lipid metabolism, but that inhibitory effects on cytokine production and inflammation may be receptor independent.


Source-sink dynamics and proteomic reprogramming under elevated night temperature and their impact on rice yield and grain quality.
High night temperatures (HNTs) can reduce significantly the global rice (Oryza sativa) yield and quality. A systematic analysis of HNT response at the physiological and molecular levels was performed under field conditions. Contrasting rice accessions, N22 (highly tolerant) and Gharib (susceptible), were evaluated at 22°C (control) and 28°C (HNT). Nitrogen (N) and nonstructural carbohydrate (NSC) translocation from different plant tissues into grains at key developmental stages, and their contribution to yield, grain-filling dynamics and quality aspects, were evaluated. Proteomic profiling of flag leaf and spikelets at 100% flowering and 12 d after flowering was conducted, and their reprogramming patterns were explored. Grain yield reduction in susceptible Gharib was traced back to the significant reduction in N and NSC translocation after flowering, resulting in reduced maximum and mean grain-filling rate, grain weight and grain quality. A combined increase in heat shock proteins (HSPs), Ca signaling proteins and efficient protein modification and repair mechanisms (particularly at the early grain-filling stage) enhanced N22 tolerance for HNT. The increased rate of grain filling and efficient proteomic protection, fueled by better assimilate translocation, overcome HNT tolerance in rice. Temporal and spatial proteome programming alters dynamically between key developmental stages and guides future transgenic and molecular analysis targeted towards crop improvement.


Incidence and diagnosis of rotavirus infection in neonates: results of two studies.
The present study investigates the incidence of human rotavirus excretion in neonates of two hospitals. From April 1, 1983 to March 31, 1984, rotaviruses were detected with Rotazyme in the feces of 57 (15%) of 383 neonates at the Department of Pediatrics, University of Berne, in a total in 71 (5.2%) of the 1369 stool samples tested. Of the 57 neonates with Rotazyme-positive stool samples, 53 (93%) were asymptomatic. The problems of the specificity of Rotazyme in neonatal feces and the possibility of rapid testing on neonatology wards are discussed. In a prospective study carried out in April 1986 at the "Schweizerische Pflegerinnenschule in Zurich", rotaviruses were detected with genome gel electrophoresis in the feces of 15 (63%) of 24 risk neonates in the premature baby ward and in 36 (46%) of 79 neonates from the infant nursery, in a total in 63 (34%) of the 188 stool samples tested. In this rotavirus outbreak all rotaviruses detected showed the same band pattern, and therefore had an identical electrophorotype. Symptoms and hospital hygiene aspects of this outbreak will be dealt with in subsequent papers.


Frequent mitotic errors in tumor cells of genetically micro-heterogeneous glioblastomas.
Glioblastoma multiforme (GBM) is characterized by intratumoral heterogeneity as to both histomorphology and genetic changes, displaying a wide variety of numerical chromosome aberrations the most common of which are monosomy 10 and trisomy 7. Moreover, GBM in vitro are known to have variable karyotypes within a given tumor cell culture leading to rapid karyotype evolution through a high incidence of secondary numerical chromosome aberrations. The aim of our study was to investigate to what extent this mitotic instability of glioblastoma cells is also present in vivo. We assessed the spatial distribution patterns of numerical chromosome aberrations in vivo in a series of 24 GBM using two-color in situ hybridization for chromosomes 7/10, 8/17, and 12/18 on consecutive 6-microm paraffin-embedded tissue slides. The chromosome aberration patterns were compared with the histomorphology of the investigated tumor assessed from a consecutive HE-stained section, and with the in vitro karyotype of cell cultures established from the tumors. All investigated chromosomes showed mitotic instability, i.e., numerical aberrations within significant amounts of tumor cells in a scattered distribution through the tumor tissue. As to chromosomes 10 and 17, only monosomy occurred, as to chromosome 7 only trisomy/polysomy, apparently as a result of selection in favor of the respective aberration. Conversely, chromosomes 8, 12, and 18 displayed scattered patterns of monosomy as well as trisomy within a given tumor reflecting a high mitotic error rate without selective effects. The karyotypes of the tumor cell cultures showed less variability of numerical aberrations apparently due to clonal adaptation to in vitro conditions. We conclude that glioblastoma cells in vivo are characterized by an extensive tendency to mitotic errors. The resulting clonal diversity of chromosomally aberrant cells may be an important biological constituent of the well-known ability of glioblastomas to preserve viable tumor cell clones under adaptive stress in vivo, in clinical terms to rapidly recur after antitumoral therapy including radio- or chemotherapy.


Pattern and outcome of surgical management of nasolachrymal duct obstruction in children: a five year review.
To determine the pattern of naso-lachrymal duct obstruction (NLDO) and outcome of dacryocystorhinostomy (DCR) in children in Aminu Kano Teaching Hospital Kano, over a 5-year period. The clinic and theater registers were used to retrieve the records of all patients below the age of 15 years who presented with NLDO. The information obtained included age, sex, duration, and types of clinical signs at presentation cause of obstruction and outcome of DCR. Successful outcome is defined as patent naso-lachrymal duct 1 year after surgery. All the patients had external DCR with stent inserted into the lachrymal sac and anchored to the columella and left in situ for 6 weeks. Fortnightly for three visits then at 2 months intervals. At each visit, the patient had lachrymal punctal cannulation and irrigation with normal saline to ensure free drainage. There were 17 patients, 9 males and 8 females (M: F = 1.1: 1). Two patients (11.8%) had bilateral disease and 15 (88.2%) were unilateral. The patients' ages ranged between 2 and 10 years. The commonest presenting features were tearing and discharge. In 14 patients (82%) tearing started from birth though patients presented much later. Congenital NLDO occurred in 82%, and in the remaining 18%, obstruction was caused by depressed nasal fracture. Fifteen patients (88%) had successful outcome of DCR at 1-year follow-up. The commonest complication noted was stent extrusion before 6 weeks in 17.6% of cases. Conclusion : NLDO was mostly of congenital origin and was characterized by delayed presentation. Patients have good outcome with external DCR.


The amino acid sequences of carboxypeptidases I and II from Aspergillus niger and their stability in the presence of divalent cations.
The amino acid sequences of serine carboxypeptidase I (CPD-I) and II (CPD-II), respectively, from Aspergillus niger have been determined by conventional Edman degradation of the reduced and vinylpyridinated enzymes and peptides hereof generated by cleavage with cyanogen bromide, iodobenzoic acid, glutamic acid cleaving enzyme, AspN-endoproteinase and EndoLysC proteinase. CPD-I consists of a single peptide chain of 471 amino acid residues, three disulfide bridges and nine N-glycosylated asparaginyl residues, while CPD-II consists of a single peptide chain of 481 amino acid residues, has three disulfide bridges, one free cysteinyl residue and nine glycosylated asparaginyl residues. The enzymes are closely related to carboxypeptidase S3 from Penicillium janthinellum. Both Ca2+ and Mg2+ stabilize CPD-I as well as CPD-II, at basic pH values, Ca2+ being most effective, while the divalent ions have no effect on the activity of the two enzymes.