Identification of a novel SBF2 missense mutation associated with a rare case of thrombocytopenia using whole-exome sequencing.
We describe in this report a case of a 6-years-old female who presented at the age of 1 month with a mucocutaneous bleeding and suspected thrombocytopenia. The patient's condition was refractory to the known idiopathic thrombocytopenic purpura treatments and congenital form of Thrombocytopenia was suspected following the delivery of a male sibling with the same phenotype. The patient also manifested fair colored hair and skin relative to her family however she did not have any detectable neurologic or other immunologic abnormalities. In order to further understand this condition, we have used whole-exome sequencing of the patient's DNA as well as her father's with the assumption that her condition is transmitted in an autosomal recessive manner. We have identified a missense change c.659C>G (p.Thr220Arg) in the SBF2 (also known as MTMR13) gene that causes a mutation in the DENN domain of the protein. This mutation was validated by traditional Sanger sequencing and analyzed in the remaining family members were it was found to segregate in the homozygous state in the affected siblings and in the heterozygous state in both parents. This novel mutation in the DENN domain of SBF2 maybe interfering with its putative association with the Rab family of small GTPases which are important mediators of vesicle transport and membrane trafficking. In conclusion, we have identified a novel mutation that is associated with severe thrombocytopenia. The fact that this mutation is found in SBF2 gene may indicate that the underlying cause of thrombocytopenia in our patient is either a new variant form of Griscelli syndrome (through the Rab GTPases action) or a variant Charcot-Marie-Tooth type 4 disease as SBF2 truncating mutations were previously identified in sufferers of this disease. This finding will help to accurately diagnose and classify similar cases of congenital thrombocytopenia and provide further proof to the power of whole-exome sequencing in personalizing patients management from the point of diagnosis to treatment and followup.


Astemizole. A review of its pharmacodynamic properties and therapeutic efficacy.
Astemizole is an H1-histamine receptor antagonist with a long duration of action permitting once daily administration. Its efficacy in seasonal and perennial allergic rhinitis has been convincingly demonstrated, and several comparative studies suggest that astemizole is at least as effective as some other H1-histamine receptor antagonists. A few smaller studies have shown beneficial effects on the symptoms of allergic conjunctivitis and chronic urticaria (but not atopic dermatitis). While astemizole appears to share with other H1-histamine receptor antagonists a tendency to increase appetite and cause weight gain after prolonged use, it offers the important advantage of an absence of significant central nervous system depression or anticholinergic effects with usual doses. Thus, astemizole offers a worthwhile improvement in side effect profile over 'traditional' H1-histamine receptor antagonists, especially in patients bothered by the sedative effects of these drugs.


The Effect of Topical Tranexamic Acid in Endoscopic Sinus Surgery: A Triple Blind Randomized Clinical Trial.
Our aim is to evaluate the effect of topical tranexamic acid (TA) on bleeding and surgical quality field in the functional endoscopic sinus surgery (FESS). A total of 74 patients who underwent FESS due to chronic rhinosinusitis were included. The patients were randomized into 2 groups. TA group (n = 37) received a topical cotton pledget soaked with TA and placebo (PL) group (n = 37) received a pledget soaked with saline solution. A significant effect was noted for the TA group versus the PL group in the grade 1 of the Boezaart scale at 35 minutes (4 for TA group and no case for PL group). This effect was absent for higher grades. We did not notice a significant effect between the 2 groups at 5 minutes. Blood loss was 359 ml in the TA group versus 441 ml in the PL group. No significant change was observed between the 2 groups concerning the blood parameters. No side effects were reported. Despite its safety when administrated locally and its low cost, TA provides limited effect on quality of surgical field after 35 minutes of the start of FESS in the patients with chronic rhinosinusitis. This effect was absent at the start of the intervention and when analyzing the blood loss and hematologic parameters.


Void filling of tibia compression fracture zones using a novel resorbable nanocrystalline hydroxyapatite paste in combination with a hydroxyapatite ceramic core: first clinical results.
It is a generally accepted standard surgical practice to fill-in the metaphyseal defect zones resulting from the reduction of tibia compression fractures. The development of various innovative bone substitutes is also currently on the increase. In our prospective study, we used Ostim, a novel resorbable nanocrystalline hydroxyapatite paste, together with Cerabone, a solid hydroxyapatite ceramic, in combination with angularly stable osteosynthesis to treat 24 tibia compression fractures. Types B2 and B3, as well as types C2 and C3 fractures, according to the AO classification, were included in the study. The mean total range of joint motion in terms of flexion and extension was improved from the immediate postoperative value of 79 +/- 14 degrees to 97 +/- 13 degrees at 6 weeks after surgery, to 109 +/- 16 degrees at 3 months, and finally to 118 +/- 17 degrees at 1 year. In three patients, a delayed wound healing was observed as a local complication. The use of the Ostim and Cerabone combination is an effective method in treating tibia compression fractures with large defect zones left after reduction.


Electrically conductive nanofibrous scaffold composed of poly(ethylene glycol)-modified polypyrrole and poly(ε-caprolactone) for tissue engineering applications.
The aim of this study was to developing two novel nanofibrous scaffolds composed of poly(ethylene glycol)-modified polypyrrole [PEG-b-(PPy)4] and poly(ε-caprolactone) (PCL) for tissue engineering (TE) applications. For this purpose, pyrrole-functionalized PEGs AB4 macromonomers (PyPEGsM) were synthesized through the Steglich esterification of PEGs ends-caped tetraol [PEGs(OH)4] using pyrrole-2-carboxylic acid. These macromonomers were subsequently copolymerized with pyrrole monomer using chemical oxidation polymerization approach to produce PEGs-b-(PPy)4 copolymers. A solution of PCL and the synthesized PEGs-b-(PPy)4 copolymers were electrospun to fabricate uniform, conductive, and biocompatible nanofibrous scaffolds. The performances of the fabricated nanofibers as TE scaffolds were examined in terms of biological (biocompatibility and biodegradability) as well as physicochemical (electroactivity, conductivity, mechanical properties, and morphology) features. As the results, the fabricated electrospun nanofibers were found as proper scaffolds for use in TE applications that require electroactivity.


Subacute skeletal myopathy induced by 2,4-dichlorophenoxyacetate in rats and guinea pigs.
Rats and guinea pigs were injected with 2,4-dichlorophenoxyacetate (2,4-D) in amounts sufficient to keep them myotonic for over 12 hr daily. The biceps brachii muscles were studied by histochemistry, epoxy resin histology, and electron microscopy after one to five days. Changes observed include proliferation of the longitudinal component of the sarcoplasmic reticulum, production of new myofilaments on the periphery of cells and in nuclei, activation of satellite cells, accumulation of lipid droplets, myofibrillar damage with Z-disc streaming, necrosis, and regeneration. The action mehanisms of 2,4-D are not well understood. Some of the phenomena observed could be classified as proliferative and may be analogous to the auxin action of 2,4-D in plants. The production of intranuclear filaments is probably related to the diffusion of newly formed cytoplasmic proteins into nuclei.


Kinetic control of phosphoglycerate mutase from Flavobacterium sp. grown on ethylene glycol.
A species of Flavobacterium able to oxidise ethylene glycol to pyruvate via glyoxylate, glycerate, 2-phosphoglycerate and phosphoenolpyruvate exploits phosphoglycerate mutase to initiate gluconeogenesis. Partially purified phosphoglycerate mutase from this bacterium is independent of adenylate charge control but is activated by phosphoenolpyruvate. The possible significance of this regulation is discussed.


Effect of acute hyperglycemia on the ischemic preconditioning effect of prodromal angina pectoris in patients with a first anterior wall acute myocardial infarction.
Acute hyperglycemia abolishes the ischemic preconditioning effect of prodromal angina pectoris in patients with acute myocardial infarction. We investigated a potential explanation for the association between acute hyperglycemia and adverse outcomes after acute myocardial infarction.


Regional variation and absence of large neurons in the cerebellum of the staggerer mouse.
Staggerer (sg) is a neurological mutant mouse in which the cerebellar granule cells degenerate after migration to the internal granule cell layer. In addition, the Purkinje cells are abnormal, being ectopic, smaller in size and without tertiary branchlet spines. In this study we report two new observations on this mutant: (1) the effects of the mutation show a regional variation in severity along the mediolateral axis. This variation is seen in the cross-sectional size of the tissue, the extent of cortical folding, as well as the density and cytological appearance of the medium-to-large cortical neurons (MLNs); (2) cell counts were done of 30-day-old mutants and of wild-type controls. The counts revealed that three-quarters of the MLNs of the cerebellar cortex are missing in staggerer. These findings cannot exclude the possibility that Golgi as well as Purkinje cells are absent since the two cannot be distinguished in staggerer. Depending on the number of Golgi cells present, between 60% and 90% of the wild-type number of Purkinje cells is missing in staggerer. The results are discussed in terms of their implications for other studies and for locating the site of staggerer gene action.


Beneficial effects of empagliflozin and liraglutide on the cerebral microcirculation of diabetic rats.
This study aimed to evaluate the effects of the antidiabetics liraglutide, a GLP-1 analog, and empagliflozin, an SGLT-2 inhibitor, on the brain microcirculation of diabetic rats. Type 2 diabetes mellitus (DM) was experimentally induced in male Wistar rats by combining a high-fat diet and a low dose of streptozotocin (35 mg/kg). Liraglutide (100 μg/kg s.c.) and empagliflozin (10 mg/kg, oral) were administered for 5 weeks. Body weight was monitored periodically. Oral glucose tolerance, fasting glycemia, and blood triglycerides were evaluated after the treatments. Endothelial-leukocyte interactions in the brain microcirculation and structural capillary density were assessed. DM rats presented metabolic and cerebrovascular alterations. Liraglutide treatment decreased body weight and blood triglycerides of DM rats. Empagliflozin treatment improved glucose tolerance but only the combination therapy significantly reduced fasting blood glucose. Both treatments and their combination reduced leukocyte adhesion into the endothelium of brain venules. However, empagliflozin was more effective in preventing DM-induced microvascular rarefaction. These findings suggest that chronic treatment with SGLT2 inhibitors and GLP-1 receptor agonists may serve as potential therapeutic approaches to prevent microvascular complications associated with diabetes.


Particle properties of sugar maple hemicellulose hydrolysate and its influence on growth and metabolic behavior of Pichia stipitis.
In this study the influence of the insoluble solids in nano-filtrated sugar maple hemicellulosic hydrolysate on the metabolic behavior of Pichia stiptis was investigated. The particle properties of hemicellulosic hydrolysate were analyzed. Phosphoric acid and ammonium (PA) were applied to remove the particles. The metabolic behavior and growth property of P. stipitis in particle--removed hydrolysate was measured. Results demonstrated that the average particle size and zeta potential of the untreated hydrolysate were 2266.9±78.2 nm and -6.09±0.49 mV. Xylose consumption and ethanol production rate were significantly decreased when particle content is greater than 1.63 g/L. Because the majority of particles (34 g/L) were removed from hydrolysates by phosphoric acid and ammonium treatment, the fermentability of the hydrolysate was significantly improved. These results indicated particles play an important role in hydrolysate inhibition effect.


Effectiveness and mechanisms of phosphate adsorption on iron-modified biochars derived from waste activated sludge.
Excessive discharge of phosphate (P) into the surface water is the key factor to cause the eutrophication, so its removal has aroused much attention in recent years. In this study, different iron modification (chemical co-precipitation of Fe3+/Fe2+ or FeCl3 impregnation) was used to improve the phosphate adsorption capacity of waste activated sludge (WAS)-based biochar. Comparative tests demonstrated that the FeCl3-impregnated WAS-based biochar exhibited much superior phosphate adsorption capacity (111.0mg/g) in all as-prepared samples and performed well even under the interferences with pH and coexisting ions. X-ray diffraction (XRD) analyzes indicated that the iron in FeCl3-impregnated WAS-based biochar existed mainly in amorphous phase, as hematite and amorphous hydroxides forms, which was of great benefit to the phosphate adsorption. Besides, ligand exchange plays important role in the adsorption of phosphate. The WAS-based biochar kept over 60% phosphate removal efficiency after five recycles.


Effects of sildenafil on blue-on-yellow and white-on-white Humphrey perimetry in 3 months regular use.
To assess the effects of sildenafil on blue-on-yellow (B/Y) and white-on-white (W/W) Humphrey perimetry in a group of men with erectile dysfunction in 3 months regular use. In this prospective study, 14 patients with erectile dysfunction received 50 mg doses of sildenafil (Viagra, Pfizer) two times per week regularly for 3 months. Patients underwent best-corrected visual acuity (BCVA), colour vision, anterior segment and fundus examination, and B/Y and W/W Humphrey perimetry in each eye before and after sildenafil treatment. Changes in mean deviations (MD) were compared separately for both eyes. No significant changes from baseline were observed in BCVA and colour vision after sildenafil treatment. The anterior segment and fundus examinations revealed no abnormalities. There was no change on B/Y and W/W Humphrey visual field. MD values of B/Y and W/W Humphrey perimetry remained nonsignificant for both eyes after 3 months regular use of sildenafil (P=0.133, and 0.092, respectively, for right, and P=0.221 and 0.925, respectively, for left eyes). Oral sildenafil used regularly for 3 months produced no effect on B/Y and W/W Humphrey perimetry.


Physiology, production and action of progesterone.
The aim of this article is to review the physiology of progesterone and focus on its physiological actions on tissues such as endometrium, uterus, mammary gland, cardiovascular system, central nervous system and bones. In the last decades, the interest of researchers has focused on the role of progesterone in genomic and non-genomic receptor mechanisms. We searched PubMed up to December 2014 for publications on progesterone/steroidogenesis. A better understanding of the biological genomic and non-genomic receptor mechanisms could enable us in the near future to obtain a more comprehensive knowledge of the safety and efficacy of this agent during hormone replacement therapy (natural progesterone), in vitro fertilization (water-soluble subcutaneous progesterone), in traumatic brain injury, Alzheimer's disease and diabetic neuropathy, even though further clinical studies are needed to prove its usefulness.


Event-Related Rumination Inventory: A Validation Process in the Greek Language.
Rumination has been identified as a negative psychological response of women diagnosed with breast cancer. The aim of the present study was to validate the Event-Related Rumination Inventory (ERRI) in Greek women with breast cancer. Sixty female patients with newly diagnosed breast cancer were included in the study. The ERRI questionnaire was translated with the back-forward procedure. Sociodemographic, anthropometric, and medical parameters were also assessed. The principal component analysis resulted in the following two-factor solution: (1) intrusive thoughts and (2) positive outcome. All subscales showed satisfactory internal consistency and variance, relative to theoretical score ranges. Subscale scores and the total score were significantly correlated with post-traumatic growth, distress, depression, and anxiety, demonstrating good criterion validity. Associations with patients' sociodemographic and medical characteristics, such as the stage of the disease and the type of treatment, were also identified. The Greek version of the ERRI provides valid and reliable measures of rumination when administered to women with breast cancer.


Patterns of physiologic and behavioral response of intermediate care preterm infants to intervention.
To determine the safety of a developmental intervention for use with preterm infants and feasibility of a time sampling procedure. An experimental design was used to test an intervention with clinically stable 33-34-week postconceptional age infants over a 4-day period. Repeated outcome measures included pulse rate, oxygen saturation levels, and infant behavioral state. The intervention resulted in significant differences in infant behavioral state with the experimental group achieving more alertness. Physiologic parameters remained within normal limits both during and after the intervention. The protocol was found to be safe for implementation with clinically stable preterm infants. Behavioral state findings suggest the potential for the intervention to promote alertness. Results suggest that changes in oxygen saturation and pulse rate were dependent upon changes in behavioral state.


Visualizing mRNA Dynamics in Live Neurons and Brain Tissues.
Localization of mRNA plays a crucial role in a variety of neuronal processes including synaptogenesis, axonal guidance, and long-term plasticity. Recent advances in fluorescence imaging and RNA labeling techniques allow us to visualize how individual mRNA molecules are dynamically regulated inside live neurons and brain tissues. Here, we describe key methods in imaging mRNA dynamics, including preparation of neuron culture and brain slices from transgenic mice expressing GFP-labeled mRNA, high-resolution detection of single molecules, live tissue imaging, and analysis of mRNA transport.


Effects of Freeze-Dried Banana and Watermelon Peel Powders on Bile Salt Resistance, Growth Kinetics, and Survival of Probiotic Bacteria.
Fruit peels have potential as prebiotic sources thanks to their dietary fiber contents. This study aimed to determine the effects of freeze-dried banana (BPP) and watermelon (WPP) peel powders on bile salt resistance, growth kinetics, and survival of Lactobacillus acidophilus and Lactiplantibacillus plantarum. In the presence of 0.5-1% bile salt, L. plantarum counts were 0.52-1.13 log CFU/mL higher in MRS broth added with 5% peel powder than without peel powder. Lactobacillus acidophilus population was 2.47-2.79 log CFU/mL higher in MRS broth added with 5% peel powder than without peel powder in the presence of 0.5% bile salt. Both peel powders did not affect the growth kinetics of L. acidophilus in milk. Conversely, the growth of L. plantarum was promoted in milk supplemented with peel powders and yielded a shorter generation time (P < 0.05). The maximum population density of L. plantarum in milk supplemented with BPP (8.68 log CFU/mL) was higher than in milk without peel powder (7.72 log CFU/mL; P < 0.05). Survival of L. acidophilus improved during storage at 4 °C in milk added with peel powders. The results suggest that BPP and WPP can be functional ingredients in probiotic foods and may be used to improve the growth and survival of probiotic cultures.


High-dose rate intracavitary therapy for carcinoma of the uterine cervix: II. Risk factors for rectal complication.
From August 1978 through December 1982, a total of 267 patients with carcinoma of the uterine cervix were treated using remote afterloading high-dose rate intracavitary therapy (RALS) with non-rigid applicator at our department. The data from 199 previously untreated patients with standard application out of 267 were available for this analysis of rectal complication. The incidence of moderate to severe rectal complication (Kottmeier's grade 2 and 3) was 7% (13/199). Cox's regression model was used for the analysis of risk factors for rectal complication in which even minor injuries (grade 1) were included. By using this method, it was clear that the first significant risk factor was z-coordinates of weighted geometric center (WGC-z) (p = 0.0007) and the second corresponding factor was rectal TDF (p = 0.0082), the sum of the rectal dose measured by semiconductor dosimeter (ICD-5) and the dose of external whole pelvic irradiation. From the analysis of application pattern of intracavitary sources, WGC-z indicated the approximation of intracavitary sources to the anterior wall of rectum quantitatively and three-dimensionally. The significance of rectal TDF also implied that the monitoring of rectal dose by ICD-5 had an important role for the prediction and prevention of rectal complication after RALS.


Association of DNA methylation of vitamin D metabolic pathway related genes with colorectal cancer risk.
Vitamin D might have anti-tumor effect, which is affected by the genes related to vitamin D metabolic pathway. Epigenetic mechanism may affect the expression level of vitamin D metabolic pathway related genes, then plays an important role in the occurrence and development of colorectal cancer. To date, no study has reported on the association between blood-based DNA methylation level of vitamin D metabolic pathway related genes and colorectal cancer risk. A case-control study was conducted including 102 colorectal cancer cases and 102 sex- and age-frequency-matched controls in Guangzhou, China. CpG islands in the VDR, CYP24A1, CYP27B1 and CYP2R1 genes were chosen for DNA methylation analysis by MethylTarget sequencing. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of DNA methylation levels for colorectal cancer. Taking the point with the largest Youden index as the boundary value, the cumulative methylation levels of vitamin D metabolic pathway related genes were divided into hypomethylation and hypermethylation. Unconditional multivariable logistical regression model was used to calculate the adjusted odds ratio (aOR) and 95% confidence intervals (95% CIs) after adjusting for potential confounders. Among 153 CpG sites, 8 CpG sites were significantly different between the cases and the controls. The cumulative methylation level of all CpG sites in CYP2R1 was inversely associated with the risk of colorectal cancer (aOR, 0.49; 95% CI, 0.26-0.91). However, no significant association was found between cumulative methylation levels of all CpG sites in VDR, CYP24A1 and CYP27B1 and colorectal cancer risk. Significant inverse association was observed between cumulative methylation level of significant CpG sites in VDR (aOR, 0.28; 95% CI, 0.16-0.51) and CYP24A1 (aOR, 0.19; 95% CI, 0.09-0.40) and colorectal cancer risk. There were no significant associations between cumulative methylation levels of significant CpG sites in CYP2R1 and CYP27B1 and colorectal cancer risk. This study indicated that the cumulative methylation levels of significant CpG sites in VDR and CYP24A1 and all CpG sites in CYP2R1 were inversely associated with colorectal cancer risk.


Fabrication and characterization of active nanocomposite films loaded with cellulose nanocrystals stabilized Pickering emulsion of clove bud oil.
There is a tremendous increase in the development of alternative food packaging materials which are functional, environment-friendly, and can improve the shelf-life of food products. One such possible approach is to develop biopolymer-based active films loaded with antimicrobial essential oils. In the present study, pearl millet starch (PMS) films reinforced with kudzu cellulose nanocrystals (CNCs) stabilized Pickering emulsions of clove bud oil (CBO) were developed as active and sustainable packaging material. Active nanocomposite films were prepared by blending PMS with Pickering emulsions of CBO at 0.5, 1, 1.5, and 2 wt% conc. Using the solution casting method. Overall, active nanocomposite films displayed improved thermal, mechanical, and water barrier properties, with an optimum CBO-Pickering emulsion concentration of 1.5 %. CBO and PMS films showed strong chemical interactions, which significantly improved the mechanical resistance of the film. Further, SEM showed the appearance of micro-porous holes in the films because of partial evaporation on the cryo-fractured surface due to the vacuum condition. In addition, films exhibited antimicrobial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), with a rate response from increasing CBO Pickering emulsion concentration from 0.5 to 2 %. E. coli and S. aureus exhibited an inhibition zone ranging from 10.5 to 2.15 mm and 11.2 to 22.1 mm. This study suggests that PMS starch and kudzu CNCs-based active nanocomposite films loaded with CBO-Pickering emulsions have good potential to develop active and sustainable packaging materials.


[The aluminum level of infant food; effect of the preparation method. Consequences in kidney dysfunction?].
Stimulated by reports in literature concerning high aluminium content of human milk formulas, ten Dutch formulas were investigated. The aluminium content appeared to be rather low. Conventional home preparation induced a pronounced rise of aluminium content (3 to 100 fold). Even in this way prepared, milk formula is safe.


Mutant isolation and cloning of the gene encoding protease VII from Escherichia coli.
A mutant of Escherichia coli lacking protease VII, the outer membrane-associated protease which specifically cleaves paired basic residues (1), was isolated by using N-methyl-N'-nitro-N-nitrosoguanidine treatment. The mutant exhibited no significant change as for its growth rate and microscopic feature compared with wild cells. The gene encoding protease VII was cloned by using complementation analysis of protease VII (-) mutation. The minicell experiment showed that the gene encoded a putative precursor protein of 38,000 Mr which was processed into a protein of 36,000 Mr suggesting the presence of a signal peptide on the putative precursor.


The diagnostic ability of 18F-FDG PET/CT for mediastinal lymph node staging using 18F-FDG uptake and volumetric CT histogram analysis in non-small cell lung cancer.
To evaluate the clinical implications of lymph node (LN) density on 18F-FDG PET/CT for mediastinal LN characterization in non-small cell lung cancer (NSCLC). One hundred and fifty-two patients with 271 mediastinal LNs who underwent PET/CT and endobronchial ultrasound-guided transbronchial needle aspiration for staging were enrolled. Maximum standardized uptake value (SUVmax), short axis diameter, LN-to-primary cancer ratio of SUVmax, and median Hounsfield unit (HU) based on CT histogram were correlated to histopathology. Of 271 nodes, 162 (59.8 %) were malignant. SUVmax, short axis diameter, and LPR of malignant LNs were higher than those of benign nodes. Among malignant LNs, 71.0 % had median HU between 25 and 45, while 78.9 % of benign LNs had values <25 HU or >45 HU. Using a cutoff value of 4.0, SUVmax showed the highest diagnostic ability for detecting malignant LNs with a specificity of 94.5 %, but showing a sensitivity of 70.4 %. Using additional density criteria (median HU 25-45) in LNs with 2.0< SUVmax ≤4.0, the sensitivity increased to 88.3 % with the specificity of 82.6 %. LN density is useful for the characterization of LNs with mild 18F-FDG uptake. The risk of mediastinal LN metastasis in NSCLC patients could be further stratified using both 18F-FDG uptake and LN density. • SUVmax showed the highest diagnostic ability for detecting malignant LNs. • LN density was useful in characterization of LNs with mild FDG uptake. • SUVmax and LN density together could stratify the risk of LN metastasis.


Ligand-specific receptor states: implications for opiate receptor signalling and regulation.
Opiate drugs produce their effects by acting upon G protein coupled receptors (GPCRs) and although they are among the most effective analgesics available, their clinical use is restricted by unwanted side effects such as tolerance, physical dependence, respiratory depression, nausea and constipation. As a class, opiates share a common profile of unwanted effects but there are also significant differences in ligand liability for producing these actions. A growing number of studies show that GPCRs may exist in multiple active states that differ in their signalling and regulatory properties and which may distinctively bind different agonists. In this review we summarize evidence supporting the existence of multiple active conformations for MORs and DORs, analyze information favouring the existence of ligand-specific receptor states and assess how ligand-selective efficacy may contribute to the production of longer lasting, better tolerated opiate analgesics.


Epithelial locomotion and differentiation in frog skin cultures.
Small explants from the medioventral skin of the green frog were maintained in culture for 5 days. During the first hours, a rapid outgrowth of the stratum germinativum was observed at the periphery of the fragment (2 X 3 X 0.75 mm). The Malpighian cells stretched and emitted long lamellipodia following the cut edges of the dermis. These cells acquired a fibroblastic shape and were covered by other flattened cells from the stratum spinosum and even from the stratum granulosum. This progression of cells simulated an epiboly; cell divisions occurred and were revealed by autoradiography. The underlying dermis could be totally covered after 3 days. An increase in the number of cellular layers was observed. The migrating cells redifferentiated, in particular at the lower side of the dermis, which provided a suitable substratum for the newly formed epidermis. A new basal lamina was formed. Some exfoliations of keratinized layers were seen. After 5 or 6 days of culture, epiboly was complete, the epithelium formed a closed system, and degenerative processes occurred, probably by non-penetration of the nutritive medium into the deeper regions of the explant.


Inhibitory influences of xanthine oxidase inhibitor and angiotensin I-converting enzyme inhibitor on multinucleated giant cell formation from monocytes by downregulation of adhesion molecules and purinergic receptors.
Allopurinol, a xanthine oxidase inhibitor, and captopril, an inhibitor of angiotensin I-converting enzyme, are widely used for hyperuricaemia and hypertension, respectively. There have been reported cases showing that these two agents are effective for the treatment of granulomatous diseases such as sarcoidosis, although the mode of action is not elucidated. We examined the in vitro effects of these agents on the formation of multinucleated giant cells (MGC) from human monocytes by concanavalin A-stimulated mononuclear cell supernatants (conditioned medium). We cultured monocytes with conditioned medium and each agent and compared the rate of MGC formation as well as the expression of adhesion molecules and P2X7 receptor, which are involved in MGC formation. The addition of 25 or 100 microg mL(-1) allopurinol or 0.125-1.0 microg mL(-1) captopril inhibited MGC formation. Monocytes treated with these agents exhibited less expression of intercellular adhesion molecular-1 (ICAM-1) than untreated monocytes. The susceptibility of monocytes cultured in conditioned medium for 24 h to 2'-and 3'-o-(4-benzoyl-benzoyl)adenosine triphosphate-mediated cytolysis was significantly lower in monocytes treated with these agents than in untreated monocytes. Allopurinol and captopril have a therapeutic effect on granulomatous disorders by a direct action on monocyte/macrophage lineage cells partly through downregulation of ICAM-1 and P2X7 receptor.


Muscle function in juvenile chronic arthritis.
Muscle strength and thickness were studied in children with juvenile chronic arthritis (JCA) to evaluate their muscle function. We studied voluntary isometric, isokinetic, and nonvoluntary isometric muscle strength, as well as muscle thickness, in 20 children with JCA. Thickness of the quadriceps muscle was measured by ultrasound. Results were compared with reference values for healthy children and a matched control group. Isometric muscle strength in knee extensors, elbow flexors, and wrist dorsiflexors was reduced in children with JCA. In muscles near an inflamed joint, the strength was 45-65% of expected value. In muscles without adjacent arthritis, the strength was slightly decreased (80-90% of expected value). Isometric and isokinetic strength in ankle dorsiflexors was reduced only in children with ankle arthritis. Nonvoluntary muscle strength in thumb adductors during electrical stimulation of the ulnar nerve was reduced in children with arthritis in the hand. Thickness of the quadriceps muscle was reduced both in children with and without knee arthritis (75 and 90% of expected). Children with JCA have reduced muscle strength and thickness, which is most pronounced in muscles near an inflamed joint.


Regulatory T cells in paraneoplastic neurological syndromes.
Focusing on CD4(+)CD25(+) regulatory T lymphocytes (T(reg)), we studied the gene expression of T(reg) functional molecules in peripheral blood lymphocytes of patients with paraneoplastic neurological syndrome (PNS), including Lambert-Eaton myasthenic syndrome (LEMS) with small cell lung carcinoma (SCLC) and anti-Hu- or anti-Yo-antibody-positive PNS. T(reg)-rich subsets were sorted from the patients' peripheral blood mononuclear cells, and the mRNA expression levels of their functional genes were measured. The expression levels of FOXP3, TGF-beta and CTLA4 mRNA in T(reg)-rich subsets of PNS patients were down-regulated compared with that of SCLC patients without PNS. These results suggest that T(reg) dysfunction plays a role in PNS development.


Enantioselective total synthesis of otteliones A and B, novel and powerful antitumor agents from the freshwater plant Ottelia alismoides.
Otteliones A and B, isolated from the freshwater plant Ottelia alismoides, have attracted significant attention because of their potential as novel anticancer agents. In this review, four independent enantioselective total syntheses and one formal synthesis of these natural products are presented with particular focus on their methodology and strategy.


On the use of D2.B10-Dmdmdx/J (D2.mdx) Versus C57BL/10ScSn-Dmdmdx/J (mdx) Mouse Models for Preclinical Studies on Duchenne Muscular Dystrophy: A Cautionary Note from Members of the TREAT-NMD Advisory Committee on Therapeutics.
The C57BL/10ScSn-Dmdmdx/J (mdx) mouse model has been used by researchers for decades as a model to study pathology of and develop therapies for Duchenne muscular dystrophy. However, the model is relatively mildly affected compared to the human situation. Recently, the D2.B10-Dmdmdx/J (D2.mdx) mouse model was suggested as a more severely affected and therefore better alternative. While the pathology of this model is indeed more pronounced early in life, it is not progressive, and increasing evidence suggest that it actually partially resolves with age. As such, caution is needed when using this model. However, as preclinical experts of the TREAT-NMD advisory committee for therapeutics (TACT), we frequently encounter study designs that underestimate this caveat. We here provide context for how to best use the two models for preclinical studies at the current stage of knowledge.


Systemic immunity against a murine colon tumor (CT-26) produced by immunization with syngeneic cells expressing a transfected viral gene product.
We have previously shown that CT-26 tumor cells expressing the transfected hemagglutination antigen of influenza virus elicit an immune response in syngeneic hosts that is cross-protective against an s.c. challenge with non-transfected parental cells. Since CT-26 is a murine colorectal carcinoma we sought to determine whether the same immuno-protective effect could be achieved after a challenge with parent tumor cells injected into the usual sites of colon tumor growth and metastasis. We now show that the protection afforded by this immunization protocol is systemic and that animals can be protected against tumor growth in the cecum, lymphatics of the mesentery, liver and lung.


Modeling pedestal experiments with amplitude instead of contrast.
To provide a comprehensive quantitative model of contrast discrimination, we measured contrast discrimination functions at four mean luminances, four spatial frequencies, three phase relations between test and pedestal gratings, and two temporal frequencies for the test grating. The results confirm previous findings that the shape of the contrast discrimination function varies with three of these variables but is independent of luminance when each discrimination threshold is divided by the detection threshold for the test grating presented alone. The data in this 5-dimensional space can be described quantitatively if expressed in amplitudes instead of contrasts. The resulting model of visual amplitude sensitivity has seven parameters that are specific to a particular observer and are tied to identifiable visual properties. The "pedestal effect" and tests with the pedestal and test gratings out-of-phase can be explained by subthreshold summation but not stimulus uncertainty.


Trace elements and carcinogenicity: a subject in review.
Cancer is known to be a multi-step process, which involves different stages including initiation, promotion, progression and metastasis. Chemical carcinogens including most trace elements can change any of these processes to induce their carcinogenic effects. Various studies confirm that cancer arises from the accumulation of irreversible DNA damage, which results from multiple mutations in critical genes in the body organ. Chemical carcinogens most often directly or after xenobiotic metabolism, act as genotoxic causes to induce DNA damage. Genotoxic carcinogen refers to a group of chemicals capable of producing cancer by directly altering the genetic material of target cells. Other carcinogens are however classified as non-genotoxic, which represents chemicals that are capable of producing cancer by some secondary mechanism not related to direct gene damage. They act as tumor promoters, endocrine-modifiers, receptor mediators, immunosuppressant, or inducers of tissue-specific toxicity and inflammatory responses. The diversity of modes of action, of non-genotoxic carcinogens, the tissue and species specificity and the absence of genotoxicity makes it extremely hard to predict their carcinogenic potential. The roles of trace metals (some of which are either genotoxic or non-genotoxic) in cancer development and inhibition have a complex character and have raised many questions because of their essential and toxic effects on people's health. Trace metals such as cadmium, nickel, arsenic, beryllium and chromium (VI) have been recognized as human or animal carcinogens by International Agency for Research on Cancer (IARC). The Carcinogenic capability of these metals depends mainly on factors such as oxidation states and chemical structures. The oxidative concept in metal carcinogenesis proposes that complexes formed by these metals, in vivo, in the vicinity of DNA, catalyze redox reactions, which in turn oxidize DNA. The most significant effect of reactive oxygen species in the carcinogenesis progression is DNA damage, which results in DNA lesions like strand breaks and the sister-chromatid exchange. This article reviews the carcinogenicity of various trace elements.


Delayed onset of graft-versus-host disease in immunodeficent human leucocyte antigen-DQ8 transgenic, murine major histocompatibility complex class II-deficient mice repopulated by human peripheral blood mononuclear cells.
Haematopoietic humanization of mice is used frequently to study the human immune system and its reaction upon experimental intervention. Immunocompromised non-obese diabetic (NOD)-Rag1(-/-) mice, additionally deficient for the common gamma chain of cytokine receptors (γc) (NOD-Rag1(-/-) γc(-/-) mice), lack B, T and natural killer (NK) cells and allow for efficient human peripheral mononuclear cell (PBMC) engraftment. However, a major experimental drawback for studies using these mice is the rapid onset of graft-versus-host disease (GVHD). In order to elucidate the contribution of the xenogenic murine major histocompatibility complex (MHC) class II in this context, we generated immunodeficient mice expressing human MHC class II [human leucocyte antigen (HLA)-DQ8] on a mouse class II-deficient background (Aβ(-/-) ). We studied repopulation and onset of GVHD in these mouse strains following transplantation of DQ8 haplotype-matched human PBMCs. The presence of HLA class II promoted the repopulation rates significantly in these mice. Virtually all the engrafted cells were CD3(+) T cells. The presence of HLA class II did not advance B cell engraftment, such that humoral immune responses were undetectable. However, the overall survival of DQ8-expressing mice was prolonged significantly compared to mice expressing mouse MHC class II molecules, and correlated with an increased time span until onset of GVHD. Our data thus demonstrate that this new mouse strain is useful to study GVHD, and the prolonged animal survival and engraftment rates make it superior for experimental intervention following PBMC engraftment.


Measurement of electrical activity of the human small intestine using surface electrodes.
Electrical activity of the human small intestine is usually measured by implanted or intraluminal electrodes. The application of these invasive techniques is, however, very limited. In this paper, a noninvasive technique is introduced to measure electrical activity of the small intestine by placing electrodes on the abdominal skin over the small intestine. Surface recordings were obtained in ten healthy volunteers, three patients with total gastrectomy and five patients with gastroparesis (a slight degree of paralysis of the mucosal coat of the stomach) with implanted electrodes on the serosa of the duodenum. An omnipresent 9-12 cpm electrical activity was observed in all surface recordings. Our findings from the surface electrodes were consistent with those reported in the literature via implanted or intubated electrodes. It is concluded that the technique described in this paper provides a noninvasive way to measure electrical activity of the small intestine. It may have potential application in medical research and clinical diagnosis.


Visualizing mineral binding and uptake of bisphosphonate by osteoclasts and non-resorbing cells.
Bisphosphonates (BPs) target bone due to their high affinity for calcium ions. During osteoclastic resorption, these drugs are released from the acidified bone surface and taken up by osteoclasts, where they act by inhibiting the prenylation of small GTPases essential for osteoclast function. However, it remains unclear exactly how osteoclasts internalise BPs from bone and whether other cells in the bone microenvironment can also take up BPs from the bone surface. We have investigated this using a novel fluorescently-labelled alendronate analogue (FL-ALN), and by examining changes in protein prenylation following treatment of cells with risedronate (RIS). Confocal microscopic analysis showed that FL-ALN was efficiently internalised from solution or from the surface of dentine by resorbing osteoclasts into intracellular vesicles. Accordingly, unprenylated Rap1A accumulated to the same extent whether osteoclasts were cultured on RIS-coated dentine or with RIS in solution. By contrast, J774 macrophages internalised FL-ALN and RIS from solution, but took up comparatively little from dentine, due to their inability to resorb the mineral. Calvarial osteoblasts and MCF-7 tumour cells internalised even less FL-ALN and RIS, both from solution and from the surface of dentine. Accordingly, the viability of J774 and MCF-7 cells was drastically reduced when cultured with RIS in solution, but not when cultured on dentine pre-coated with RIS. However, when J774 macrophages were co-cultured with rabbit osteoclasts, J774 cells that were adjacent to resorbing osteoclasts frequently internalised more FL-ALN than J774 cells more distant from osteoclasts. This was possibly a result of increased availability of BP to these J774 cells due to transcytosis through osteoclasts, since FL-ALN partially co-localised with trancytosed, resorbed matrix protein within osteoclasts. In addition, J774 cells occupying resorption pits internalised more FL-ALN than those on unresorbed surfaces. These data demonstrate that osteoclasts are able to take up large amounts of BP, due to their ability to release the BP from the dentine surface during resorption. By contrast, non-resorbing cells take up only small amounts of BP that becomes available due to natural desorption from the dentine surface. However, BP uptake by non-resorbing cells can be increased when cultured in the presence of resorbing osteoclasts.


High-frequency haplotypes in the X chromosome locus TLR8 are associated with both CD and UC in females.
TNF-alpha and IL-1 have been associated with mucosal inflammation in both Crohn's disease (CD) and ulcerative colitis (UC). Innate immune defects have been associated with CD, specifically CARD15/NOD2. Recently, Toll-like receptor 8 (TLR8) signaling has been shown to enhance generation of both cytokines. Interestingly, TLR8 is located on the X chromosome and inflammatory bowel disease (IBD) has been associated with abnormalities of the X chromosome. The aim was to test whether TLR8 haplotypes are associated with IBD. Subjects (735 CD, 343 UC, 245 controls) were genotyped. Single nucleotide polymorphisms (SNPs) were chosen to tag common Caucasian haplotypes. Both "risk (H4)" and "protective (H1)" TLR8 haplotypes were observed associated with CD in females. Eighteen percent of CD females had H4 compared with 9% of controls (P = 0.02). Fifty-nine percent of CD females had H1 compared with 72% of controls (P = 0.01). H1 was also negatively associated with UC in females (59% of UC, 72% of controls P = 0.03). Diplotype analysis of CD, UC, and all IBD in females revealed that 2 protective haplotypes (H1/H1) had a markedly diminished odds ratio, 0.4-0.5. The presence of a risk haplotype (H4 / not H1) had a significantly increased odds ratio, 2.0-2.2. Thus, the risk for IBD was 4-5 times higher in females with 1 risk haplotype than with the protective/protective diplotype. TLR8 is an X-linked IBD susceptibility gene with both common predisposing and protecting haplotypes. These associations further emphasize the importance of genetic variation in innate immunity as determinants, not only of CD, but of UC as well.


8-Oxo-7,8-dihydrodeoxyadenosine: the first example of a native DNA lesion that stabilizes human telomeric G-quadruplex DNA.
Native DNA lesions in general destabilize DNA secondary structures such as duplex and G-quadruplex because they disrupt optimized interactions in DNA defined by nature. In this paper, we report the first example of a native DNA lesion (8-oxo-7,8-dihydrodeoxyadenosine, OxodA) that stabilizes human telomeric G-quadruplex DNA. CD thermal denaturation studies explicitly displayed increased melting temperatures of telomeric G-quadruplex DNAs that contain OxodA(s) in different DNA loops, suggesting enhanced thermal stability. Conformation studies of G-quadruplex DNAs containing OxodA(s) in the loops using CD and native PAGE revealed that they adopt a similar antiparallel conformation in Na(+) but have much more versatile conformations in K(+). According to computational calculations, the observed stabilization may result from the tight binding of K(+) into the pocket formed by the O8 of OxodA and its loop. The study reported here may provide better understanding of the effect of DNA lesions on G-quadruplex stability and conformation.


Design and immunological properties of Helicobacter pylori glycoconjugates based on a truncated lipopolysaccharide lacking Lewis antigen and comprising an α-1,6-glucan chain.
To investigate the vaccine potential of H. pylori lipopolysaccharide (LPS), truncated LPS of H. pylori strain 26695 HP0826::Kan lacking O-chain polysaccharide and comprising an extended α-1,6-linked glucan chain was conjugated to tetanus toxoid (TT) or bovine serum albumin (BSA). Two approaches were used for delipidation or partial delipidation of H. pylori LPS: (1) mild hydrolysis resulting in delipidated LPS (dLPS) and (2) treatment with anhydrous hydrazine resulting in removal of O-linked fatty acids (LPS-OH). Both LPS-OH and dLPS were covalently linked through a 2-keto-3-deoxy-octulosonic acid (Kdo) residue to a diamino group-containing spacer, followed by conjugation to thiolated TT or BSA to give conjugates LPS-OH-TT, dLPS-BSA and dLPS-TT, respectively. The LPS-OH-TT, dLPS-BSA and dLPS-TT conjugates were immunogenic in both rabbits and mice, inducing strong and specific IgG responses against homologous and heterologous strains of H. pylori. Moreover, the rabbit post-immune sera showed cross-reactivity against clinical isolates of H. pylori in a whole-cell indirect ELISA, which was further confirmed by indirect immunofluorescent microscopy. A tenfold stronger IgG immune response to the immunizing antigen was generated in mice and rabbits that received dLPS-containing conjugate. The post-immune sera of rabbits immunized with LPS-OH-TT, dLPS-BSA or dLPS-TT displayed significant bactericidal activity against mutant and wild-type α-1,6-glucan-expressing strains and selected clinical isolates of H. pylori. Finally, partial protection against H. pylori challenge was demonstrated in mice vaccinated with dLPS-TT conjugate adjuvanted with cholera toxin. In summary, this study shows that glycoconjugates based on delipidated or partially delipidated LPS from H. pylori 26695 HP0826::Kan mutant induce broadly cross-reactive functional antibodies in immunized animals and should be considered for further vaccine development and testing.


Novel mutation in the gene encoding c-Abl-binding protein SH3BP2 causes cherubism.
Cherubism is a rare autosomal dominant inherited condition caused by mutations in the c-Abl-binding protein SH3BP2. It is characterized by multiple cystic giant cell lesions of the jaw appearing in early childhood with stabilization and remission after puberty. In the present study, we used direct sequence analysis of the SH3BP2 gene of several individuals from a family with cherubism to search for additional SH3BP2 mutations resulting in cherubism. In affected relatives, we found a previously unreported G to A transition in exon 9 leading to a Gly to Arg substitution at amino acid position 420. G420R has been reported previously with a G to C transversion. To date there have been no disease causing mutations outside exon 9. Therefore, the amino acid sequence from positions 415 to 420 may represent a specific protein domain which, when disrupted, leads to the cherubism phenotype.


Validation of semi-automatic device UA-704 for self-measurement of blood pressure.
To perform a clinical validation for a semi-automatic arm-type device UA-704 (A&D Company, Ltd., Tokyo, Japan) according to the British Hypertension Society protocol. Eighty-five study participants (37 men) were included. The mean age was 50.1+/-17.0 years, systolic blood pressure range was 79-212 mmHg and diastolic blood pressure range was 43-118 mmHg. For each participant, three readings of the UA-704 were compared with simultaneous auscultatory measurements by two trained independent observers. The observers used a mercury calibrated sphygmomanometer and a dual stethoscope. The results were graded according to the 1993 British Hypertension Society protocol. The average difference between mercury sphygmomanometer and UA-704 readings was -1.85+/-4.26 mmHg (mean+/-SD) for systolic blood pressure and -1.44+/-3.97 mmHg for diastolic blood pressure. The proportions of values agreeing to within 5, 10 and 15 mmHg were 79.2%, 96.5% and 99.6% for systolic blood pressure and 86.7%, 96.9% and 99.6% for diastolic blood pressure for the observers and device (A/A grade for British Hypertension Society). For an adult population, the UA-704 device for self-measurement of blood pressure achieved a British Hypertension Society grade A/A and therefore can be recommended for home blood pressure monitoring.


Optimization of culture conditions for the expression of three different insoluble proteins in Escherichia coli.
Recombinant protein expression for structural and therapeutic applications requires the use of systems with high expression yields. Escherichia coli is considered the workhorse for this purpose, given its fast growth rate and feasible manipulation. However, bacterial inclusion body formation remains a challenge for further protein purification. We analyzed and optimized the expression conditions for three different proteins: an anti-MICA scFv, MICA, and p19 subunit of IL-23. We used a response surface methodology based on a three-level Box-Behnken design, which included three factors: post-induction temperature, post-induction time and IPTG concentration. Comparing this information with soluble protein data in a principal component analysis revealed that insoluble and soluble proteins have different optimal conditions for post-induction temperature, post-induction time, IPTG concentration and in amino acid sequence features. Finally, we optimized the refolding conditions of the least expressed protein, anti-MICA scFv, using a fast dilution protocol with different additives, obtaining soluble and active scFv for binding assays. These results allowed us to obtain higher yields of proteins expressed in inclusion bodies. Further studies using the system proposed in this study may lead to the identification of optimal environmental factors for a given protein sequence, favoring the acceleration of bioprocess development and structural studies.


Neonatal jaundice in 'healthy' very low birthweight infants.
The daily bilirubin levels during the first week of life in 94 premature very low birthweight (VLBW, less than 1500 g) relatively 'healthy' infants were determined. Mean daily bilirubin values peaked on the fourth day of life at 188.1 mumol/l (s.e.m. = 5.3). Twenty-eight infants developed hyperbilirubinaemia (bilirubin greater than 260 mumol/l), at which time they were exposed to phototherapy. When individual peak bilirubin values were evaluated, the overall peak value was 213.9 mumol/l (s.e.m. = 5.1) occurring at 4.81 days (s.e.m. = 0.11), although the value is most likely an underestimate, since the 28 pre-phototherapy values were not truly peak values. Seventy-six (81%) infants experienced bilirubin levels greater than 170 mumol/l. The method of delivery apparently had no impact on the bilirubin levels. All the infants remained well and progressed satisfactorily. 'Healthy' VLBW infants experience a much greater incidence and severity of neonatal jaundice than mature infants with the same clinical status.


Carbon pathways during transitory starch degradation in Arabidopsis differentially affect the starch granule number and morphology in the dpe2/phs1 mutant background.
The Arabidopsis knockout mutant lacking both the cytosolic disproportionating enzyme 2 (DPE2) and the plastidial phosphorylase (PHS1) had a dwarf-growth phenotype, a reduced and uneven distribution of starch within the plant rosettes, and a lower starch granule number per chloroplast under standard growth conditions. In contrast, a triple mutant impaired in starch degradation by its additional lack of the glucan, water dikinase (GWD) showed improved plant growth, a starch-excess phenotype, and a homogeneous starch distribution. Furthermore, the number of starch granules per chloroplast was increased and was similar to the wild type. We concluded that ongoing starch degradation is mainly responsible for the observed phenotype of dpe2/phs1. Next, we generated two further triple mutants lacking either the phosphoglucan, water dikinase (PWD), or the disproportionating enzyme 1 (DPE1) in the background of the double mutant. Analysis of the starch metabolism revealed that even minor ongoing starch degradation observed in dpe2/phs1/pwd maintained the double mutant phenotype. In contrast, an additional blockage in the glucose pathway of starch breakdown, as in dpe2/phs1/dpe1, resulted in a nearly starch-free phenotype and massive chloroplast degradation. The characterized mutants were discussed in the context of starch granule formation.


Circulating thrombomodulin as a novel endothelial cell marker: comparison of its behavior with von Willebrand factor and tissue-type plasminogen activator.
Circulating thrombomodulin is a novel endothelial cell marker, which may reflect the endothelial injury. Plasma levels of thrombomodulin were quantitated by an enzyme-linked immunosorbent assay (ELISA) in patients with hematological malignancies, liver disease, diabetes mellitus, collagen disease, thrombotic disease, and disseminated intravascular coagulation (DIC), and the thrombomodulin values were compared with those of von Willebrand factor antigen (vWf:Ag) and tissue-type plasminogen activator (t-PA) which are released from stimulated or damaged endothelial cells. The mean plasma concentrations of thrombomodulin in these disease states were elevated as compared with healthy subjects. A relatively high mean thrombomodulin level was observed in DIC, liver disease, and collagen disease. Abnormally high thrombomodulin values (greater than normal mean value + 3 SD) were found in 32.3% of patients with hematological malignancies, 57.7% of patients with liver disease, 39.3% of patients with diabetes mellitus, 30.0% of patients with collagen disease, 23.1% of patients with thrombotic disease, and 69.0% of patients with DIC. Plasma concentrations of both vWf:Ag and t-PA were also elevated in these patients. On the whole, the plasma thrombomodulin concentration was positively correlated with vWf:Ag (r = 0.441, P less than 0.001) and t-PA (r = 0.398, P less than 0.001). These findings indicate that the elevation of plasma thrombomodulin is frequently seen in a variety of diseases and circulating thrombomodulin is possibly useful for evaluating the endothelial damage in selected disease states.


Usefulness of the platelet-to-lymphocyte ratio in predicting the severity of carotid artery stenosis in patients undergoing carotid angiography.
Carotid artery stenosis (CAS) is primarily caused by atherosclerotic plaque. Progressive inflammation may contribute to the rupture of an atherosclerotic plaque. The platelet-to-lymphocyte ratio (PLR) is a new and simple marker that indicates inflammation. In this study, we aimed to investigate the use of the PLR to determine the severity of CAS. One hundred forty patients were chosen from among patients who underwent carotid angiography in our institution. Symptomatic patients with stenosis >50% in the carotid arteries and asymptomatic patients with stenosis >80% were diagnosed via carotid angiography as having critical stenosis. Patients were classified into two groups. Group 1 included patients who had critical CAS, whereas Group 2 included patients with noncritical CAS, as determined by carotid angiography. Correlations between the PLR and the severity of CAS were analyzed. There were no significant differences in sex and age between the two groups. The PLR was 162.5 ± 84.7 in the noncritical CAS group patients and 94.9 ± 60.3 in the critical CAS group patients (p < 0.0001). The PLR value of 117.1 had 89% sensitivity and 68% specificity for CAS [95% confidence interval, 0.043-0.159; area under the curve, 0.101 ± 0.03)]. In this study, we have shown that PLR values may be associated with critical stenosis in at least one of the carotid arteries. Furthermore, PLR values may be used to predict critical stenosis in the carotid arteries.


Novel acridine-based LSD1 inhibitors enhance immune response in gastric cancer.
Recently, histone lysine specific demethylase 1 (LSD1) has become an emerging and promising target for cancer immunotherapy. Herein, based on our previously reported LSD1 inhibitor DXJ-1 (also called 6x), a series of novel acridine-based LSD1 inhibitors were identified via structure optimizations. Among them, compound 5ac demonstrated significantly enhanced inhibitory activity against LSD1 with an IC50 value of 13 nM, about 4.6-fold more potent than DXJ-1 (IC50 = 73 nM). Molecular docking studies revealed that compound 5ac could dock well into the active site of LSD1. Further mechanism studies showed that compound 5ac inhibited the stemness and migration of gastric cancer cells, and reduced the expression of PD-L1 in BGC-823 and MFC cells. More importantly, BGC-823 cells were more sensitive to T cell killing when treated with compound 5ac. Besides, the tumor growth was also suppressed by compound 5ac in mice. Together, 5ac could serve as a promising candidate to enhance immune response in gastric cancer.


Comment on 'Therapeutic application of metallic nanoparticles combined with particle-induced x-ray emission effect'.
A recent paper (Kim et al 2010 Nanotechnology 21 425102) presented results on the combination of irradiation by atomic ions of cells loaded by particles made of heavy atoms. They propose that the projectile induced x-rays emission (PIXE) mechanism has an important contribution to the enhancement of the cell death rate. Experiments made in our group to study the effects of such a combination have shown that the Auger effect induced in the high-Z atoms and the following induction of surrounding water radiolysis has an important contribution to the enhancement of the cell death rate. In the light of our studies we propose an alternative interpretation of the results presented in the paper by Kim et al.


Is the in situ accessibility of the 16S rRNA of Escherichia coli for Cy3-labeled oligonucleotide probes predicted by a three-dimensional structure model of the 30S ribosomal subunit?
Systematic studies on the hybridization of fluorescently labeled, rRNA-targeted oligonucleotides have shown strong variations in in situ accessibility. Reliable predictions of target site accessibility would contribute to more-rational design of probes for the identification of individual microbial cells in their natural environments. During the past 3 years, numerous studies of the higher-order structure of the ribosome have advanced our understanding of its spatial conformation. These studies range from the identification of rRNA-rRNA interactions based on covariation analyses to physical imaging of the ribosome for the identification of protein-rRNA interactions. Here we reevaluate our Escherichia coli 16S rRNA in situ accessibility data with regard to a tertiary-structure model of the small subunit of the ribosome. We localized target sequences of 176 oligonucleotides on a 3.0-A-resolution three-dimensional (3D) model of the 30S ribosomal subunit. Little correlation was found between probe hybridization efficiency and the proximity of the probe target region to the surface of the 30S ribosomal subunit model. We attribute this to the fact that fluorescence in situ hybridization is performed on fixed cells containing denatured ribosomes, whereas 3D models of the ribosome are based on its native conformation. The effects of different fixation and hybridization protocols on the fluorescence signals conferred by a set of 10 representative probes were tested. The presence or absence of the strongly denaturing detergent sodium dodecyl sulfate had a much more pronounced effect than a change of fixative from paraformaldehyde to ethanol.


1,3,4, Oxadiazole Compound A3 Provides Robust Protection Against PTZ-Induced Neuroinflammation and Oxidative Stress by Regulating Nrf2-Pathway [Retraction].
[This retracts the article DOI: 10.2147/JIR.S333451.].


Body Composition Impacts Hematopoietic Stem Cell Transplant Outcomes in Both Autologous and Allogeneic Transplants: A Systematic Review.
According to World Health Organization, over 50,000 hematopoietic stem cell transplants (HSCTs) are performed annually worldwide. Though HSCT can extend life-expectancy and improve disease-related health burdens, it is not without risks. Post-transplant overall survival is improving; therefore, it is imperative that factors contributing to or impeding further improvements are well understood. The purpose of this systematic review is to explore current data on body composition (specifically weight loss, BMI, obesity and sarcopenia) and the relation to HSCT outcomes. A literature search was conducted via PubMed and Web of Science databases. Key words included "body composition," "sarcopenia," "hematopoietic stem cell transplant," "malnutrition," "body mass index," and "obesity." Results indicated that 16 out of 18 analyzed studies found a statistically significant relationship between body composition, in particular higher BMI and weight loss, and at least one survival-related outcome variable (eg., non-relapse mortality, overall survival and/or relapse). Based on the findings of this review, body composition, whether evaluated before or during HSCT, can impact a wide variety of post-transplant outcomes. This speaks to the importance of evaluating patients pre-transplant, identifying potential risk factors for worsened outcomes, and providing immediate interventions in order to optimize transplant outcomes.


Purulent meningitis due to aspergillosis in a patient with systemic lupus erythematosus.
We report a 39-year-old female patient with systemic lupus erythematosus under immunosuppressive therapy who developed persistent neutrophilic meningitis, for which no infectious agent could be identified. Intensifying the immunosuppressive therapy induced a short amelioration of the clinical picture. At autopsy, basal meningitis was found to be due to Aspergillus sp.


Mouse ovarian factors with angiogenic activities: partial characterization.
Extracts of nonluteal mouse ovaries (Jcl: ICR strain) were assayed for neovascularization by implanting Elvax films, impregnated with test samples, on the lateral wall of the sheath of m. rectus abdominis in young adult mice of the same strain. Neovascularization occurred in a dose-dependent manner. Angiogenic activity was increased in extracts of ovaries from mice treated with follicle-stimulating hormone (FSH). The angiogenic activity was retained when the extract was heated to 100 degrees C for 15 min or adsorbed with activated charcoal. The ovarian extract was purified by affinity chromatography on a Con A-Sepharose 4B column. The adsorbed fraction possessed higher angiogenic activity. The ovarian extract was fractionated by ammonium sulfate precipitation. High angiogenic activity was found in the fraction collected between 20 to 50% saturation.


Medical workforce issues in Australia: "tomorrow's doctors--too few, too far".
The Australian medical workforce, like those of most developed countries, is increasingly "feminised" and exposed to the global market for doctors. Demand for healthcare services is increasing in the Australian community. Concern in relation to doctor shortages is increasing, particularly in rural areas. There should be greater flexibility for entry of highly-trained overseas doctors. There is an urgent need to increase medical school student intake. Issues of workforce practice, including "task" substitution, should be explored.


[Detecting infectious agents in central nervous system inflammation by cerebrospinal fluid analysis].
Cerebrospinal fluid analysis is the method of choice in CNS infection and provides the basis for appropriate treatment. Due to the proximity of CSF and CNS, the infectious agent may be detected directly by microscopy or antigen or nucleic acid detection--the latter by polymerase chain reaction--in native CSF or after culture. Furthermore, intrathecal antibody synthesis against the infectious agent may identify the cause of infection. This indirect antigen detection method requires correction for a systemic antibody response and a blood-CSF barrier disturbance. The following text gives an overview of appropriate detection methods and their relevance to the most important CNS infections.


Pyridine nucleotide oxidation-reduction state of the cerebral cortex in the awake gerbil.
A new approach to study the effect of ischemia on the brain of the awake gerbil is described. The measurement of NADH fluorescence from the surface of the cortex is done by a time-sharing fluorometer/reflectometer connected to the brain via a flexible light guide and an implanted cannula. The response of the gerbil brain to anoxia and spreading depression is described. By unilateral occlusion of the carotid artery an increase in NADH was measured in the ipsilateral hemisphere.


1-[2-(2-Benzoyl- and 2-benzylphenoxy)ethyl]uracils as potent anti-HIV-1 agents.
Non-nucleoside reverse transcriptase inhibitors (NNRTI) are key components in highly active antiretroviral therapy for treating HIV-1. Herein we present the synthesis for a series of N1-alkylated uracil derivatives bearing ω-(2-benzyl- and 2-benzoylphenoxy)alkyl substituents as novel NNRTIs. These compounds displayed anti-HIV activity similar to that of nevirapine and several of them exhibited activity against the K103N/Y181C RT mutant HIV-1 strain. Further evaluation revealed that the inhibitors were active against most nevirapine-resistant mono- and di-substituted RTs with the exception of the V106A RT. Thus, the candidate compounds can be regarded as potential lead compounds against the wild-type virus and drug-resistant forms.


The relationship between different forms of human alpha-mannosidase.
The tissue distribution and some properties of human alpha-mannosidase (alpha-D-mannoside mannohydrolase EC 188.8.131.52) have been studied. The acidic forms of the enzyme were fairly stable, whereas the neutral forms easily lost enzymic activity. The acidic forms were sensitive to neuraminidase but the neutral forms were unaffected. The experiments indicate that the acidic components are closely related to each other, differing only in sialic acid content and possibly conformation. The neutral forms of the enzyme are probably quite different from the acidic forms both in structure and cellular function.


Mutations in axonemal dynein assembly factor DNAAF3 cause primary ciliary dyskinesia.
Primary ciliary dyskinesia most often arises from loss of the dynein motors that power ciliary beating. Here we show that DNAAF3 (also known as PF22), a previously uncharacterized protein, is essential for the preassembly of dyneins into complexes before their transport into cilia. We identified loss-of-function mutations in the human DNAAF3 gene in individuals from families with situs inversus and defects in the assembly of inner and outer dynein arms. Knockdown of dnaaf3 in zebrafish likewise disrupts dynein arm assembly and ciliary motility, causing primary ciliary dyskinesia phenotypes that include hydrocephalus and laterality malformations. Chlamydomonas reinhardtii PF22 is exclusively cytoplasmic, and a PF22-null mutant cannot assemble any outer and some inner dynein arms. Altered abundance of dynein subunits in mutant cytoplasm suggests that DNAAF3 (PF22) acts at a similar stage as other preassembly proteins, for example, DNAAF2 (also known as PF13 or KTU) and DNAAF1 (also known as ODA7 or LRRC50), in the dynein preassembly pathway. These results support the existence of a conserved, multistep pathway for the cytoplasmic formation of assembly competent ciliary dynein complexes.


School adjustment of children from rural migrant families in urban China.
The purpose of this study was to examine school adjustment of rural-to-urban migrant children and its relations with acculturation in China. Migrant children were those whose official hukou status was in a rural region outside the city. Data were collected for 1175 students (M age = 11 years) in urban public schools from multiple sources including peer evaluations, teacher ratings, self-reports, and school records. The results showed that migrant students performed more competently than urban non-migrant students in social and academic areas. Migrant students displayed better psychological adjustment than non-migrant students in higher grades, but not in lower grades. Among migrant students, those with higher scores on accommodation to urban culture and maintenance of rural culture tended to be better adjusted. These results indicate the implications of migration and change in life circumstances for children's school functioning in social, academic, and psychological domains.


The effect of PINK1/Parkin pathway on glucose homeostasis imbalance induced by tacrolimus in mouse livers.
Treatment using the immunosuppressive drug tacrolimus (TAC) is related to new-onset diabetes after transplantation (NODAT). Previous studies focused mainly on islet β cells in the diabetogenic effect of TAC. Herein, we revealed that NODAT was probably induced by TAC via hepatic insulin resistance. After daily injection of mice with TAC, a glucose metabolism disorder was induced. In addition, TAC decreased the mRNA and protein levels of insulin receptor substrate 2 (IRS2), glucose transporter type 2 (GLUT2), and the phosphorylation of protein kinase B beta (pAKT2), which indicated impaired hepatic insulin signaling. Furthermore, the PTEN-induced novel kinase 1(PINK1)/Parkin pathway was shown to have a key role in the TAC-induced imbalance of hepatic glucose homeostasis. Mechanistic investigations in human hepatic cell lines revealed that TAC stimulated PINK1/Parkin expression and inhibited the expression of insulin signaling related molecules (e.g., IRS2, GLUT2 and pAKT2). Knockdown of hepatic PINK1 regulated downstream molecules of the PINK1/Parkin pathway (GLUT2 and IRS2), which reversed TAC-induced insulin resistance. Thus, in the liver, PINK1/Parkin signaling plays an important role in the TAC-induced imbalance of glucose homeostasis. TAC-induced diabetes might be prevented using Targeted treatment.


Serial changes in soluble triggering receptor expressed on myeloid cells in the lung during development of ventilator-associated pneumonia.
To determine the diagnostic role of soluble triggering receptor expressed on myeloid cells (sTREM)-1 in non-directed bronchial lavage fluid in ventilator-associated pneumonia (VAP). Non-directed bronchial lavage fluid and plasma were collected on alternate days in critically ill mechanically ventilated patients from the start of ventilatory support until complete weaning from the ventilator. Soluble TREM-1 levels were measured by an enzyme-linked immunosorbent assay. A general adult medical and surgical university hospital intensive care unit. Nine patients who developed VAP and 19 patients who did not develop VAP (controls). Plasma levels of sTREM-1 did not change significantly in either patient group. While in controls concentrations of sTREM-1 in non-directed bronchial lavage fluid did not change significantly over time, in patients who developed VAP levels of sTREM-1 in non-directed bronchial lavage fluid increased towards the diagnosis of VAP. A cut-off value for non-directed bronchial lavage fluid sTREM-1 levels of 200 pg/ml on the day of VAP had a diagnostic sensitivity of 75% and a specificity of 84%. Sensitivity increased when taking into account all sTREM-1 levels higher than 200 pg/ml from the 6-day period before the day of diagnosis that were preceded by an increase of at least 100 pg/ml (sensitivity 88%, specificity 84%). Soluble TREM-1 is a potential biomarker of VAP.


The role of DNA base excision repair in filamentation in Escherichia coli K-12 adhered to epithelial HEp-2 cells.
Base excision repair (BER) is dedicated to the repair of oxidative DNA damage caused by reactive oxygen species generated by chemical and physical agents or by metabolism which can react with DNA and cause a variety of mutations. Epithelial cells are typically the first type of host cell to come into contact with potential microbial invaders. In this work, we have evaluated whether the adherence to human epithelial cells causes DNA damage and associated filamentation. Experiments concerning adherence to HEp-2 cells were carried out with mutants deficient in BER that were derived from Escherichia coli K-12. Since the removal of mannose during bacterial interaction with HEp-2 cells allows adhesion through mannose-sensitive adhesins, the experiments were also performed in the presence and the absence of mannose. Our results showed enhanced filamentation for the single xth (BW9091) and triple xth nfo nth (BW535) mutants in adherence assays with HEp-2 cells performed without D: -mannose. The increased filamentation growth was inhibited by complementation of BER mutants with a wild type xth gene. Moreover, we measured SOS induction of bacteria adhered to HEp-2 cells in the presence and absence of D: -mannose through of SOS-chromotest assay and we observed a higher β-galactosidase expression in the absence of mannose. In this context, data showed evidence that bacterial attachment to HEp-2 epithelial surfaces can generate DNA lesions and SOS induction.


Modulation of the homophilic interaction between the first and second Ig modules of neural cell adhesion molecule by heparin.
The second Ig module (IgII) of the neural cell adhesion molecule (NCAM) is known to bind to the first Ig module (IgI) of NCAM (so-called homophilic binding) and to interact with heparan sulfate and chondroitin sulfate glycoconjugates. We here show by NMR that the heparin and chondroitin sulfate-binding sites (HBS and CBS, respectively) in IgII coincide, and that this site overlaps with the homophilic binding site. Using NMR and surface plasmon resonance (SPR) analyses we demonstrate that interaction between IgII and heparin indeed interferes with the homophilic interaction between IgI and IgII. Accordingly, we show that treatment of cerebellar granule neurons (CGNs) with heparin inhibits NCAM-mediated outgrowth. In contrast, treatment with heparinase III or chondroitinase ABC abrogates NCAM-mediated neurite outgrowth in CGNs emphasizing the importance of the presence of heparan/chondroitin sulfates for proper NCAM function. Finally, a peptide encompassing HBS in IgII, termed the heparin-binding peptide (HBP), is shown to promote neurite outgrowth in CGNs. These observations indicate that neuronal differentiation induced by homophilic NCAM interaction is modulated by interactions with heparan/chondroitin sulfates.


Investigation of Ochratoxin A biosynthetic genes inPenicillium verrucosum by DDRT-PCR experiments: differential expression of OTA genes.
A defined minimal medium has been developed which is either restrictive or permissive for the production of ochratoxin A byPenicillium verrucosum simply by changing the nitrogen and carbon source. The combination of ammonium/glycerin promoted, whereas the combination nitrate/glucose repressed ochratoxin production.In parallel mutants ofP verrucosum have been constructed by restriction endonuclease mediated integration, which were not able to produce ochratoxin. Different types of transformants, which either produce no detectable amounts of ochratoxin A but ochratoxin α, strains which produce only ochratoxin B and strains which produces none of these secondary metabolites, have been observed.


Understanding nursing assessment of health literacy in a hospital context: A qualitative study.
Nurses are fundamental in presenting information to their patients to ensure understanding of information, and health directions enabling improved health outcomes. Limited research exists exploring how nurses assess patient's health literacy in Australia. To explore Australian nurses' perceptions of patients' health literacy and how nurses assess health literacy to provide patient education. A qualitative study underpinned by phenomenology. Registered Nurses (N = 19) across five Queensland hospitals participated in semi-structured interviews which explored nurses' assessment of patient's health literacy level and their practices of delivering education. The transcripts were analysed using an inductive process with interpretative analysis. Four themes were identified: how we assess patient health literacy; challenges with health literacy assessment; consumer focused assessment; and building assessment strategies. Participants relied on cues from the patient to identify when information has not been understood. Participants felt that online training programs in the workplace would assist in education for assessment techniques; identification of patients with low health literacy; and how to best communicate with patients that have low health literacy. Formal health literacy assessments should be introduced to hospitals in Australia, but training is required to enable the nurse, the confidence and ability for health literacy assessment. Tailored education based on health literacy assessment would enhance the patient's understanding and improve discharge planning, which may reduce health service costs and readmissions. The Consolidated criteria for reporting qualitative research (COREQ) guidelines for qualitative research were followed. Registered Nurses (N = 19) participated in qualitative interviews, providing data for analysis. This study shows that nurses are already conducting informal assessments, simply by using observation and looking for cues. More education for nurses about health literacy and how to tailor their discussions with patients will improve communication.


Postheparin plasma diamine oxidase in health and intestinal disease.
In animals, the distribution of the enzyme diamine oxidase is confined, almost exclusively, to the small bowel mucosa. In humans, plasma diamine oxidase is at or below assay detection limits but can be liberated into the circulation from binding sites in the intestine by i.v. heparin. Therefore, the authors wished to see if diamine oxidase could be released by a low and safe dose of heparin (5000 U) and if the resultant area under the concentration-time curve would provide a noninvasive marker of segmental intestinal disease. In 17 control subjects, the mean area under the curve (following administration of 5000 U i.v. heparin) was 35.9 +/- 5.0 (SEM) mU.L-1.2 h-1; in 6 individuals studied on two separate occasions, postheparin plasma diamine oxidase profiles were reproducible (r = 0.98; p less than 0.001). The longitudinal distribution of diamine oxidase in the gastrointestinal tract, measured in 12 gastric, 16 jejunal, 6 ileal, and 18 colonic biopsies, was similar in humans to that found in animals. In patients with normal peroral biopsies, there was a linear relationship between jejunal mucosal and postheparin plasma diamine oxidase activities (r = 0.84; p less than 0.01). The areas under the curve in controls were then compared with those in patients with segmental intestinal diseases: 21 with ileal disease with or without colonic Crohn's disease (10 unoperated and 11 with ileal resection), 7 with non-Crohn's ileal resection, 8 with ulcerative colitis, 10 with untreated and 7 with treated celiac disease (6 studied before and after a gluten-free diet), and 5 studied during total parenteral nutrition and again after resumption of oral feeding. The results in the 18 ileectomized patients were subdivided into those with major (arbitrarily greater than 75 cm) and minor (less than 75 cm) resections. Areas under the curve were markedly reduced in nonresected Crohn's patients (6.0 +/- 1.79 mU.L-1.2 h-1; p less than 0.001 vs. controls), correlating inversely, in a first-order relationship, with disease activity (r = 0.82; p less than 0.001) and returning toward normal in 4 patients achieving disease remission. Low areas under the curve in total parenteral nutrition patients (4.5 +/- 0.9; p less than 0.001) were also reversible on resumption of oral feeding. However, areas under the curve were not significantly lower in patients with limited ileal resection (less than or equal to 75 cm), with celiac disease (untreated and treated), or ulcerative colitis than in controls.(ABSTRACT TRUNCATED AT 400 WORDS)


Leprosy: clinical and immunopathological characteristics.
Leprosy, a disease caused by Mycobacterium leprae, has polymorphic neurocutaneous manifestations strongly correlated with the host immune response. Peripheral neural damage can lead to sensory and motor losses, as well as deformities of the hands and feet. Both innate and acquired immune responses are involved, but the disease has been classically described along a Th1/Th2 spectrum, where the Th1 pole corresponds to the more limited presentations and the Th2 to the multibacillary ones. The aim of this review is to discuss this dichotomy in light of the current knowledge of the cytokines, T helper subpopulations, and regulatory T cells involved in each presentation of leprosy. The text will also address leprosy reactions related to increased inflammatory activity in both limited and multibacillary presentations, leading to exacerbation of chronic signs and symptoms and/or the development of new ones. Despite the efforts of many research groups around the world, there is no standardized serological test/biological marker for diagnosis so far, even in endemic areas, which could contribute to the eradication of leprosy.


Optimal information disclosure strategy in the primary healthcare service market: From the perspective of signaling theory.
The promotion of general practitioner (GP) contract service is one of the key components of China's healthcare reform. We consider GPs providing primary health services with private competency information over two periods, where patients decide when to sign. Two types of GPs are considered: those with higher and lower competency. Under asymmetric information, to spur the patients' incentive to sign, the GPs can move to offer competency disclosure schemes to patients, for example, separating or pooling, through which true competency information is revealed, respectively. We investigate three scenarios, which are referred to as "separating-separating," "pooling-separating," and "pooling-pooling." The results of the three scenarios yield intriguing insights into the impact of the GP's competency disclosure decisions. Findings include that GPs prefer the "pooling-separating" strategy, but patients prefer "separating-separating." Besides, an extremely low cure rate may enable GPs to conceal some competency information. Furthermore, low-competency GPs may exaggerate their competency level for profit, but greater efforts in disclosing competency information may result in diminished benefits. Therefore, to promote the services of GPs, the core is always to improve GPs' competency.


Complex regional pain syndrome (CPRS type I) after a burn injury of the hand.
Complex regional pain syndrome (CRPS) type I is a clinical condition characterized by persistent pain in one part or the entire extremity after a minor trauma, fracture, or after an operation which does not involve nerve damage and/or sympathetic hyperactivity. Despite large-scale studies on the complications that arise after burns, literature reveals few reports on neurological problems and CRPS developing after burns. It is a rare complication of a burn injury to an extremity. Its early signs and symptoms are similar to those of burn wound itself. This study describes an unusual cause of complex regional pain syndrome in burn patients. The report highlights physical examination findings, the new diagnostic criteria of complex regional pain syndrome, and difficulties in diagnosis.


The effect of dietary protein deficiency on albumin synthesis and on the concentration of active albumin messenger ribonucleic acid in rat liver.
In rats fed on a protein-deficient diet, albumin synthesis as a percentage of total liver protein synthesis falls from the normal value of approx. 15% to about 8%. We have extracted total cytoplasmic RNA from individual rat livers and measured the concentration of active albumin mRNA by translation in a reticulocyte lysate system from which the endogenous mRNA had been removed [Pelham & Jackson (1976) Eur. J. Biochem. 67, 247-256]. In this messenger-dependent system it is possible to measure the synthesis of albumin as a proportion of the overall protein synthesis promoted by the addition of the hepatic RNA. The results show that the concentration of translatable albumin mRNA in samples of total cytoplasmic RNA from livers of protein-deficient rats is decreased markedly. These findings suggest that dietary protein supply affects selectively the synthesis and/or functional stability of albumin mRNA in rat liver.


Intracellular manipulation of chromatin using magnetic nanoparticles.
Magnetic tweezers are widely used for manipulating small magnetic beads inside the cell cytoplasm in order to gain insight into the structural and mechanical properties of the cytoskeleton. Here we discuss the use of magnetic tweezers for the study of nuclear architecture and the mechanical properties of chromatin in living cells. A custom-built, dedicated micro magnetic tweezer set-up is described. We review progress that has been made in applying this technology for the study of chromatin structure and discuss its prospects for the in situ analysis of nuclear architecture and chromatin function.


Prognostic significance of SSEP, BAEP and serum S-100B monitoring after aneurysm surgery.
Changes in evoked potentials (EPs) and increased levels of S-100B protein were used to identify cerebral ischemia or glial damage and to predict neurological outcome in aneurysm patients. Somatosensory evoked potentials and Brainstem auditory-evoked potentials, and serum S-100B protein were simultaneously investigated pre- and postoperatively over a period of 10 days in 43 patients with 47 aneurysms (six in the posterior fossa). The EP scores showed a strong correlation with the clinical outcome. Sensitivity was 73%, and specificity 81%. Pathological S-100B levels >0.5 mg/l were equal in predictive values (correct positive eight, false positive six, correct negative 26, false negative three). Initially increased S-100B levels, long-lasting S-100B elevation, and secondary increasing S-100B values correlated with an unfavorable outcome. High peak S-100B values correlated with bad EP scores at discharge. EP deterioration was the first indicator anticipating S-100B elevation and clinical deterioration in five patients. There was a good correlation between pathological S-100B values or EP findings and infarction on CT scan. Both EPs and S-100B protein showed a comparable high predictive value for outcome. S-100B reflects the extent of primary brain damage after subarachnoid hemorrhage and time course of ongoing secondary brain damage. Evoked potentials assess the functional integrity and tended to react earlier than S-100B protein before definitive structural damage occurred.


Effects of laser irradiation (670-nm InGaP and 830-nm GaAlAs) on burn of second-degree in rats.
This study investigated the effects of 670-nm indium gallium phosphide (InGaP) and 830-nm gallium aluminum arsenide (GaAlAs) laser therapy on second-degree burns induced on the back of Wistar rats. Sixty-three male Wistar rats were anesthetized, and second-degree burns were made on their back. The animals were then divided randomly into three groups: control (C), animals treated with 670-nm InGaP laser (LIn), and animals treated with 830-nm GaAlAs laser (LGa). The wound areas were removed after 2, 6, 10, 14, and 18 days of treatment and submitted to structural and morphometric analysis. The following parameters were studied: total number of granulocytes and fibroblasts, number of newly formed blood vessels, and percentage of birefringent collagen fibers in the repair area. Morphometric analysis showed that different lasers 670-nm InGaP and 830-nm GaAlAs reduced the number of granulocytes and an increase of newly formed vessels in radiated lesions. The 670-nm InGaP laser therapy was more effective in increasing the number of fibroblasts. The different treatments modified the expression of VEGF and TGF-β1, when compared with lesions not irradiated. The different types of light sources showed similar effects, improved the healing of second-degree burns and can help for treating this type of injury. Despite the large number of studies with LLTI application in second-degree burns, there is still divergence about the best irradiation parameters to be used. Further studies are needed for developing a protocol effective in treating this type of injury.


Do Bayley-III Composite Scores at 18-22 Months Corrected Age Predict Full-Scale IQ at 6-7 Years in Children Born Extremely Preterm?
To determine the ability of the Bayley-III cognitive and language composite scores at 18-22 months corrected age to predict WISC-IV Full Scale IQ (FSIQ) at 6-7 years in infants born extremely preterm. Children in this study were part of the Neuroimaging and Neurodevelopmental Outcome cohort, a secondary study to the SUPPORT trial and born 240/7-276/7 weeks gestational age. Bayley-III cognitive and language scores and WISC-IV FSIQ were compared with pairwise Pearson correlation coefficients and adjusted for medical and socioeconomic variables using linear mixed effect regression models. Bayley-III cognitive (r = 0.33) and language scores (r = 0.44) were mildly correlated with WISC-IV FSIQ score. Of the children with Bayley-III cognitive scores of <70, 67% also had FSIQ of <70. There was less consistency for children with Bayley-III scores in the 85-100 range; 43% had an FSIQ of <85 and 10% an FSIQ of <70. Among those with Bayley-III language scores >100, approximately 1 in 5 had an FSIQ of <85. A cut point of 92 for the cognitive composite score resulted in sensitivity (0.60), specificity (0.64). A cut point of 88 for the language composite score produced sensitivity (0.61), specificity (0.70). Findings indicate the Bayley-III cognitive and language scores correlate with later IQ, but may fail to predict delay or misclassify children who are not delayed at school age. The Bayley-III can be a useful tool to help identify children born extremely preterm who have below average cognitive scores and may be at the greatest risk for ongoing cognitive difficulties. Extended Follow-up at School Age for the SUPPORT Neuroimaging and Neurodevelopmental Outcomes (NEURO) Cohort: NCT00233324.


Multiple autophosphorylation sites of the epidermal growth factor receptor are essential for receptor kinase activity and internalization. Contrasting significance of tyrosine 992 in the native and truncated receptors.
The role of epidermal growth factor (EGF) receptor autophosphorylation sites in the regulation of receptor functions has been studied using cells transfected with mutant EGF receptors. Simultaneous point mutation of 4 tyrosines (Y1068, Y1086, Y1148, Y1173) to phenylalanine, as well as removal of these sites by truncation of the carboxyl-terminal 123 amino acid residues, resulted in reduced receptor phosphorylation of an in vivo specific substrate phospholipase C-gamma 1 to less than 50% compared to the wild-type receptor. The internalization rate constant Ke was also significantly lower in these mutants (0.15/min) compared to cells transfected with wild-type receptor (0.27/min). Additional mutation of tyrosine 992 to phenylalanine in the truncated receptor mutant (Dc-123F) further decreased the receptor internalization rate to a minimal level (ke = 0.07-0.10/min), equivalent to the ke measured for cells expressing kinase-negative receptor (A721). Moreover, tyrosine kinase activity of the Dc-123F receptor toward phospholipase C-gamma 1, compared to wild-type receptor, was reduced by 90%. Taken together, these results show that EGF receptor lacking five autophosphorylation sites functions similar to a kinase-negative receptor. Mutation of tyrosine residue Y992 alone in the context of full length EGF receptor, however, did not affect receptor internalization or kinase activity toward phospholipase C-gamma 1. These data indicate that tyrosine 992 is critical for substrate phosphorylation and internalization only in the context of the truncated receptor, and that minor autophosphorylation sites, such as Y992, may act as compensatory regulatory sties in the absence of the major EGF receptor autophosphorylation sites.


Acute depletion of human core nucleoporin reveals direct roles in transcription control but dispensability for 3D genome organization.
The nuclear pore complex (NPC) comprises more than 30 nucleoporins (NUPs) and is a hallmark of eukaryotes. NUPs have been suggested to be important in regulating gene transcription and 3D genome organization. However, evidence in support of their direct roles remains limited. Here, by Cut&Run, we find that core NUPs display broad but also cell-type-specific association with active promoters and enhancers in human cells. Auxin-mediated rapid depletion of two NUPs demonstrates that NUP93, but not NUP35, directly and specifically controls gene transcription. NUP93 directly activates genes with high levels of RNA polymerase II loading and transcriptional elongation by facilitating full BRD4 recruitment to their active enhancers. dCas9-based tethering confirms a direct and causal role of NUP93 in gene transcriptional activation. Unexpectedly, in situ Hi-C and H3K27ac or H3K4me1 HiChIP results upon acute NUP93 depletion show negligible changesS2211-1247(22)01437-1 of 3D genome organization ranging from A/B compartments and topologically associating domains (TADs) to enhancer-promoter contacts.