Neuropeptide-Y enhances luteinizing hormone (LH)-releasing hormone-induced LH release and elevations in cytosolic Ca2+ in rat anterior pituitary cells: evidence for involvement of extracellular Ca2+ influx through voltage-sensitive channels.
The present studies were designed to investigate the mechanism by which neuropeptide-Y (NPY) augments the effect of LHRH to stimulate the release of LH from cultured rat anterior pituitary cells. Anterior pituitary cells from ovariectomized rats were enzymatically dispersed, cultured for 3 days, and then exposed to various secretagogues during 3-h incubations. As reported by this laboratory previously, NPY alone (100 nM) did not affect LH release, but significantly enhanced the LH response to 1 nM LHRH. This facilitatory action of NPY was mimicked by the dihydropyridine Ca2+ channel agonist Bay K 8644 (1 microM), and the enhancement of LHRH-induced LH release by either NPY or Bay K 8644 was prevented by the dihydropyridine antagonist nitrendipine (1 microM). Nitrendipine alone reduced the response to LHRH by approximately 25%, but did not affect basal LH release. In contrast, NPY failed to amplify the release of PRL in response to TRH, another Ca2(+)-mobilizing hormone. To test whether NPY also enhances the increase in cytosolic Ca2+ induced by LHRH, anterior pituitary cells were acutely dispersed into single cell suspensions, loaded with the fluorescent Ca2+ probe Indo-1 AM, and analyzed with a UV laser in an EPICS-753 flow cytometer at a rate of 500 cells/sec for 200 sec. The ratio of intracellular fluorescence resulting from Ca2+ bound to the Indo-1 to the fluorescence from Indo-1 alone (Indo-1 ratio), which is an index of the concentration of free cytosolic Ca2+, was determined for each cell. Approximately 7% of anterior pituitary cells responded to LHRH (1 or 10 nM) with significant increases in Indo-1 ratios, indicative of an increase in the concentration of free cytosolic Ca2+. EGTA (2.5 mM) reduced the basal Indo-1 ratios and attenuated, but did not abolish, the initial increase in response to LHRH, consistent with the initial extracellular Ca2+ influx-independent phase of the response to LHRH. NPY alone (100 nM) did not affect the Indo-1 ratios in anterior pituitary cells, but pretreatment with the peptide for 10 min before the scans significantly augmented the Indo-1 ratio response to 10 nM LHRH. This effect of NPY was also blocked by EGTA. Taken together, these biochemical and pharmacological studies suggest that NPY enhances the release of LH stimulated by LHRH by increasing extracellular Ca2+ entry, possibly by selectively affecting that component of the response involving dihydropyridine-sensitive L-type voltage-sensitive Ca2+ channels during the initial stages of the cellular response to LHRH.


Expression of glutamate transporter GLAST in the developing mouse cochlea.
The immunohistochemical localization of glutamate transporter GLAST in the developing mouse cochlea was studied at different ages between 0 and 30 days after birth (DAB). In the adult mouse cochlea, intense GLAST-like immunoreactivity was found in the supporting cells adjacent to the inner hair cells of the organ of Corti, the type II and suprastrial fibrocytes of the cochlear lateral wall, the fibrocytes of the spiral limbus and the satellite cells surrounding the spiral ganglion cells. At 0 DAB, weak GLAST-like immunoreactivity was found in the supporting cells around the immature inner hair cells. Immature fibrocytes in the cochlea were also positively immunostained. At 3 DAB, weak immunostaining of GLAST appeared in the immature satellite cells in the spiral ganglion. The GLAST-like immunoreactivity in the supporting cells around the inner hair cells, in the fiborocytes in the spiral ligament and the spiral limbus and in the satellite cells in the spiral ganglion increased progressively during the second postnatal week, and reached the adult level at 15 DAB. This time course correlates with the electrophysiological onset and maturation of the mouse auditory function, which is mediated by glutamatergic neurotransmission. These results suggest that the expression of GLAST may be needed for the efficient removal and metabolism of the released glutamate in the cochlea and may play important roles in the onset and maturation of the auditory system.


Structural characterization of ionic gelator studied by dynamic light scattering and small-angle neutron scattering.
Structural characterization of a hydrogel consisting of an oligomeric electrolyte, poly[pyridinium-1,4-diyliminocarbonyl-1,4-phenylenemethylene chloride] (1-Cl) as an ionic gelator was carried out by static and dynamic light scattering (SLS/DLS) and small-angle neutron scattering (SANS) techniques. All the measurements were performed by changing the concentration and temperature. We have successfully obtained the weight average molecular weight and the degree of polymerization of poly(1-Cl) by SLS. The sol-gel transition was clearly observed as large fluctuations in the scattering intensity of the time-intensity correlation function. Time correlation function of the scattering light intensity entailed a power law behavior at the sol-gel transition. 1-Cl hydrogel showed strong hysteresis and its hysteresis loop was observed both by DLS and theological methods. We have estimated the critical concentration of gelation and the gelation temperature by DLS. The enthalpy change for gelation was estimated to be ca. -10 kJ/mol. SANS experiments revealed that the unit structure of the gel network is responsible for the gelation of 1-Cl hydrogel.


Balance of the prooxidant and antioxidant system is associated with mortality in critically ill patients.
It is well known that oxidative stress causes certain diseases and organ damage. However, roles of oxidative stress in the acute phase of critical patients remain to be elucidated. This study aimed to investigate the balance of oxidative and antioxidative system and to clarify the association between oxidative stress and mortality in critically ill patients. This cohort study enrolled 247 patients transported to our emergency department by ambulance. Blood was drawn on hospital arrival, and serum derivatives of reactive oxidant metabolites (dROMs, oxidative index) and biological antioxidant potential (BAP, antioxidative index) were measured. Modified ratio (MR) is also calculated as BAP/dROMs/7.51. There were 197 survivors and 50 non-survivors. In the non-survivors, dROMs were significantly lower (274 vs 311, p<0.01), BAP was significantly higher (2,853 vs 2,138, p<0.01), and MR was significantly higher (1.51 vs 0.92, p<0.01) compared to those in the survivors. The AUC of MR was similar to that for the APACHE II score. Contrary to our expectations, higher BAP and lower dROMs were observed on admission in non-survivors. This may suggest that the antioxidative system is more dominant in the acute phase of severe insults and that the balance toward a higher antioxidative system is associated with mortality.


Endocrine treatment of advanced breast cancer.
Endocrine treatment is the preferred first-line therapy for advanced breast cancer because of its good tolerability. There are several groups of agents which differ in their mode of action but almost all in some way reduce stimulation by oestrogen. Over recent years, there has been a substantial increase in the number of medical agents in clinical trial. New antioestrogens with reduced or no agonist activity, and potent, more specific, aromatase inhibitors are now reaching the clinic. In most cases, the clinical effectiveness of these agents remains to be fully established and their optimal therapeutic application is yet to be determined.


Emerging issues in disorders/differences of sex development (DSD).
Disorders/Differences of Sex Development (DSD), as defined by the 2006 Consensus Statement, are "congenital conditions in which development of chromosomal, gonadal, or anatomic sex is atypical." They represent a spectrum of chronic medical conditions collectively affecting about 1% of the population and are associated with increased risk of infertility, cancer, and psychosocial distress. Clinical management in DSD is subject to multiple controversies about gender assignment, the timing and appropriateness of genital surgery and the approach to disclosure. There is dissent within and between stakeholders (healthcare providers, advocacy groups, families) regarding what constitutes optimal care. This special issue investigates the progress made as well as the uncertainties remaining a decade after the consensus statement and the gaps to be filled by future research and improved clinical practice. It discusses the increasing intricacy of genetic variant interpretation in the era of next-generation sequencing and the associated complexity of phenotypic variability. The issue tackles ethical dilemmas and the complicated decision-making process of assignment of sex of rearing at birth in cases of 5-alpha reductase type 2 deficiency, surveys delivery of clinical services in the United States, discusses challenges of interdisciplinary care and of educating patients and parents about DSD,and reviews the factors predisposing to gonadal tumor and their consequences on clinical management.


Genetics of red body polymorphism in the isopod, Venezillo evergladensis.
A red body morph with reddish-orange body and brownish-red eyes is described in the terrestrial isopod, Venezillo evergladensis. The alternate, non-red morph, has a brown body and black eyes and either gray or orange antennae. Breeding results show three alleles, rg, ro and r at an autosomal locus control the variation. The dominance hierarchy is rg less than ro less that r; the genotypes, rg/rg, rg/ro, and rg/r are non-red, grey, ro/ro and ro/r are non-red, orange, and r/r are red. The gray antennae of non-red, gray individuals is variable in expression although still objectively recognized. The red body morph and its inheritance is paralleled in related species, while the antennal variation identifies a new allele. Sperm storage, exceeding a full year, and multiple matings in nature are documented.


Indications for selective decontamination of the digestive tract.
Controversy exists as to the utility of selective decontamination of the digestive tract (SDD) as a method of infection prevention in critically ill patients. A number of prospective randomly controlled studies and 2 meta-analyses have shown a statistically significant protective effective of SDD against nosocomial infection. Most other SDD trials have shown reductions in nosocomial infections, but these reductions have not been statistically significant. Nearly all SDD studies have shown a less clear mortality benefit, and in no trial has cost-effectiveness been critically evaluated. Side effects of SDD have been limited. Despite nearly a decade of use at some institutions, reports of antimicrobic resistance evolving with SDD and causing disease are infrequent. At this time, SDD cannot be recommended for all intensive care patients in all clinical settings. SDD may be an effective infection prevention method in intensive care units in which there is a high rate of nosocomial pneumonia and/or other infections and in the postoperative period of orthotopic liver transplantation. SDD may also be useful to eliminate resistant gram-negative bacilli colonizing patients who are at high risk for infection sequelae. When SDD is used, periodic surveillance for the emergence of resistant microorganisms is imperative.


The importance of good record-keeping.
When large numbers of health care staff from different professional groups are involved in the care of individual clients, the need for simple, understandable, uniform casenotes becomes paramount. This article describes how a survey of the different casenotes used by various CPN teams led to the devising of a new system. The aim was to encourage good practice and develop a standardised record-keeping system.


Treatment of gingival recession with collagen membranes.
Periodontal plastic surgical techniques to obtain root coverage traditionally have utilized autogenous soft tissue grafts. Guided tissue regeneration (GTR) increasingly is being used as an alternative to soft tissue grafting in attempting root coverage. Resorbable membranes in GTR therapy prevent the need for a second surgery to harvest donor tissue. GTR membranes composed of collagen are effective barriers that add to the thickness of the overlying gingiva and possess platelet-aggregating properties that may help in early wound healing. The cases presented here demonstrate the treatment of gingival recession with a resorbable collagen membrane.


Pentoxifylline promotes replication of human cytomegalovirus in vivo and in vitro.
OKT3 monoclonal antibody (MoAb) therapy is well established in the prevention and therapy of acute rejection in transplant patients. Unfortunately, this therapy is associated with several short-term (cytokine release syndrome) and long-term (infections, EBV-related lymphoma) side effects. Recently, we were able to demonstrate an association between the TNF alpha release following the first OKT3 MoAb infusions and the appearance of human cytomegalovirus (HCMV) reactivation several days later. In order to prevent this TNF alpha associated HCMV reactivation patients were additionally treated with pentoxifylline (PTX), a methylxanthine derivative that has been shown to suppress TNF alpha induction. Although the TNF alpha peak plasma level following OKT3 MoAb treatment was markedly reduced, the incidence of HCMV reactivation and HCMV disease was not influenced. In transient transfection experiments using HCMV immediate early enhancer/promoter CAT reporter gene constructs PTX enhanced the promoter activity independently from TNF alpha in premonocytic cells. Furthermore, PTX acted synergistically with TNF alpha. In virus-infected human embryonal lung fibroblasts HCMV replication was triggered in the presence of both PTX and TNF alpha, while either substance alone had only marginal effects. The stimulatory effect of PTX on the immediate early (IE) enhancer/promoter was mediated via CREB/ ATF, a eukaryotic transcription factor that binds to the 19 bp sequence motif in the enhancer region, while TNF alpha stimulation was mediated by activation of the transcription factor NF-kappaB and its binding to the 18 bp sequence motif in the enhancer. These data suggest a potential side effect of cAMP-elevating drugs such as PTX.


The binding mechanism between azoles and FgCYP51B, sterol 14α-demethylase of Fusarium graminearum.
Fusarium graminearum is the main pathogen of Fusarium head blight (FHB), a worldwide plant disease and a major disease of wheat in China. Control of FHB is mainly dependent on the application of demethylase inhibitor (DMI) fungicides. Fungal sterol 14α-demethylase enzymes (CYP51) are the main target for DMI fungicides. A molecular modeling study and biological evaluation were performed to investigate the binding mechanism between azoles and CYP51B in F. graminearum. A homology model based on the crystal structure of Aspergillus fumigatus was built. Molecular docking and molecular dynamics (MD) simulations were then used to identify the optimum binding mode of propiconazole (PRP), diniconazole (DIN), triadimenol (TRL), tebuconazole (TEC) and triadimefon (TRN) with FgCYP51B. Furthermore, the binding free energy of the five protein-inhibitor complexes was calculated using molecular mechanics generalized Born surface area and Poisson-Boltzmann surface area (MM-GB/PBSA) methods. Key residues in the selective binding of azoles to FgCYP51B were recognized by per-residue free energy decomposition analysis. The five ligands have a similar binding mode in the active pocket. The binding free energy to the enzyme for inhibitors PRP and TEC is more favorable than that of TRN, TRL and DIN. Furthermore, the amino acid residues Phe511, Val136, Ile374, Ala308, Ser312 and Try137 of FgCYP51B are key residues interacting with azoles fungicides. From the experimental evaluation, the 50% effective concentration (EC50 ) values for PRP, TEC, DIN, TRL and TRN are 0.024, 0.047, 0.148, 0.154 and 0.474 mg L-1 , respectively. These five molecules exhibit potential inhibitory activity against CYP51B protein from F. graminearum. Azole fungicides for FgCYP51B should possess more hydrophobic groups interacting with residues Phe511, Val136, Ile374, Ala308, Ser312 and Tyr137. PRP and TEC are preferable for the control of FHB than DIN, TRL and TRN. © 2017 Society of Chemical Industry.


A Ligamentous Agony: Median Arcuate Ligament Syndrome as an Under-Recognized Cause of Abdominal Pain.
Median arcuate ligament (MAL) syndrome (MALS) is a rare clinical entity characterized by chronic abdominal pain resulting from compression of the celiac artery by the MAL. We present a case of MALS with imaging evidence of anterior compression of the celiac artery on expiration, which was relieved on inspiration. A 33-year-old woman presented with intermittent upper abdominal pain since three months. The pain was associated with nausea, abdominal bloating and diarrhea. Physical examination revealed a palpable abdominal aorta with no bruit. Abdominal sonogram, upper and lower gastrointestinal endoscopies, celiac disease screening, clostridium difficile toxin assays and hepatobiliary iminodiacetic acid scan were all normal. Computerized tomography angiogram (CTA) revealed subtle narrowing at the origin of the celiac artery without any atherosclerosis or calcification. Lateral aortic angiography showed anterior impression on the celiac artery at its origin on expiration. The constriction was relieved on inspiratory film - findings most consistent with MALS. MALS is rare, typically presenting with non-specific symptoms including nausea, vomiting, chronic post-prandial abdominal pain or unintentional weight loss. Angiography with respiratory maneuvers remains the diagnostic standard. However, non-invasive vascular imaging during both phases of respiration can be considered as an initial diagnostic test. The primary goal of therapy is celiac artery decompression with the additional objective of neuronolysis of the celiac ganglion. Our case highlights that MALS should be considered as a differential diagnosis in chronic, recurrent abdominal pain, particularly with an unrevealing initial evaluation. Evidence of celiac artery compression on vascular imaging, with characteristic respiratory variation, is highly suggestive of MALS.


Metabolism of dehydroepiandrosterone sulfate and estrone-sulfate by human platelets.
The aim of the present research was to study the uptake of DHEAS, and to establish the intracrine capacity of human platelets to produce sex steroid hormones. The DHEAS transport was evaluated through the uptake of [(3)H]-DHEAS in the presence or absence of different substrates through the organic anion transporting polypeptide (OATP) family. The activity of sulfatase enzyme was evaluated, and the metabolism of DHEAS was measured by the conversion of [(3)H]-DHEAS to [(3)H]-androstenedione, [(3)H]-testosterone, [(3)H]-estrone and [(3)H]-17beta-estradiol. Results indicated the existence in the plasma membrane of an OATP with high affinity for DHEAS and estrone sulphate (E(1)S). The platelets showed the capacity to convert DHEAS to active DHEA by the steroid-sulfatase activity. The cells resulted to be a potential site for androgens production, since they have the capacity to produce androstenedione and testosterone; in addition, they reduced [(3)H]-estrone to [(3)H]-17beta-estradiol. This is the first demonstration that human platelets are able to import DHEAS and E(1)S using the OATP family and to convert DHEAS to active DHEA, and to transform E(1)S to 17beta-estradiol.


BAP31 Regulates Wnt Signaling to Modulate Cell Migration in Lung Cancer.
B-cell receptor-associated protein 31 (BAP31) has been shown to overexpress in a wide range type of cancers. The present study aims to investigate the role of BAP31 on migration in lung cancer. Results showed that the migration of BAP31 knockdown cells was weaken than the control cells. Applying TGFβ to treat BAP31 knockdown cells could reduce cell migration. The enhancement on proliferation by TGFβ treatment was downregulated after BAP31 knockdown. The cell death and G0/G1 phase arrest was increased in the cells with TGFβ and BAP31 siRNA treatment when compared with TGFβ treatment alone. Gene expression analysis showed that Bax/Bcl2, MLKL and LC3 was upregulated in the cells with combinatorial treatment of TGFβ and BAP31 siRNA. In addition, BAP31 was shown to regulate multiple signaling pathways, especially for Wnt signaling. It found that BAP31 knockdown cells treated with TGFβ decreased β-catenin cytosolic expression and nuclear localization. Wnt signaling activator BIO could restore the downregulation of proliferation by BAP31 knockdown. This finding suggested that BAP31 regulated cancer cell migration is possibly involved with cell death mechanisms and Wnt signaling.


Effect of cytostatic agents on salivary gland tissue of the rat. The influence of 5-fluorouracil.
Application of 5-fluorouracil (5FU) for 3 weeks considerably decreased submaxillary gland weight but had no apparent influence on amylase and protein concentrations in gland extracts and isoamylase distribution patterns as revealed by electrophoresis. After another 3 weeks during which no 5FU was administered, an overproduction of protein synthesis (especially amylase) was observed.


Kinetic studies on the aggregation of Aspergillus niger conidia.
Morphology has a crucial effect on productivity and the supply of substrate for cultures of filamentous fungi. However, cultivation parameters leading to the desired morphology are often chosen empirically as the mechanisms governing the processes involved are usually unknown. For coagulating microorganisms like Aspergillus niger the morphological development is considered to start with the aggregation of conidia right after inoculation. To elucidate the mechanism of this process, kinetic studies were carried out using an in-line particle size analyzer. Based on the data obtained from these experiments a model for conidial aggregation is proposed in this article. It consists of two separate aggregation steps. The first one takes place immediately after inoculation, but only leads to a small decrease of total particle concentration. Most suspended conidia aggregate after a second aggregation step triggered by germination and hyphal growth. Aggregation velocity of this second phase is linearly dependent on the particle growth rate.


On the issue of closed versus open forms of gamma-aminobutyric acid (GABA) in water: Ab initio molecular dynamics and metadynamics studies.
Gamma-aminobutyric acid (GABA), a primary neurotransmitter, accomplishes its activities by binding to different receptor sites in different conformations. It is known to have two major conformers: the closed and open forms. Earlier studies on preferred conformation of GABA in water revealed differing results with some reporting the open form while others inferring the closed form to be more stable. We found the existence of many open forms and only one closed form of GABA in water through ab initio metadynamics simulation. Some of the open conformers are equally or more stable while others are less stable than the closed form. Free energy barriers reveal that different conformers are interconvertible at room temperature in typical experimental time scales. Ab initio molecular dynamics simulations are performed to further investigate the inter-conversion of various conformers of GABA in water and their dipole moments and also to make connections to experiments on the conformation of GABA in water.


Idiopathic hypertrophic cranial pachymeningitis: an unusual cause of subacute and chronic headache.
Three cases of idiopathic hypertrophic cranial pachymeningitis are presented. The diagnosis was based on the CT scan or MRI findings (or both) of thickened enhancing dura. In all cases, meningeal biopsies were performed and microscopic findings were compatible with nonspecific inflammation. All cases presented with subacute and chronic localized headache. Two cases had associated chronic meningitis. One cases presented with a syndrome of multiple cranial nerve involvement (polyneuritis cranialis). Corticosteroids, in the form of prednisolone 60 mg/day, were effective in all cases. Two cases with less severe pachymeningitis received corticosteroids for 2 weeks, then were tapered off in 4 to 6 weeks. A case with extensive lesions needed a long-term low dosage of prednisolone, 5 to 10 mg/day for maintenance therapy. Idiopathic hypertrophic cranial pachymeningitis may be related to the Tolosa-Hunt syndrome, the syndrome of polyneuritis cranialis, and multifocal fibrosclerosis.


The role of the periosteum in the healing of the dog's Müller-plate treated femur fracture.
There are contradictory opinions of the role of the periosteum in the conditions of stable osteosynthesis. One group of authors (Küntscher, 1970; Zucman et al., 1968) and others believe that the periosteum, especially its cambian layer, has one of the most important roles in the healing of fractures. However, other group of authors (McEven, 1912; Antony et al., 1980) and others deny the crucial role of the periosteum in the healing of fractures. The authors of the present study performed osteosynthesis by the Müller plate in both femoral bones of 20 dogs. In the left femoral bone, the periosteum was removed 4 cm in the proximal and distal direction from the fracture site, while in the right femoral bone the periosteum remained intact. The healing of the fracture was controlled by X-ray every 15 days up to 2.5 months when the animals were subjected to euthanasia. After 2.5 months from the introduction of the fracture, the healing of the fracture line was recorded in both femoral bones. It is concluded that in the conditions of stable osteosynthesis the periosteum does not play a crucial role in the healing of bones.


Clostridium acetobutylicum 8-oxoguanine DNA glycosylase (Ogg) differs from eukaryotic Oggs with respect to opposite base discrimination.
During repair of damaged DNA, the oxidized base 8-oxoguanine (8-oxoG) is removed by 8-oxoguanine-DNA glycosylase (Ogg) in eukaryotes and most archaea, whereas in most bacteria it is removed by formamidopyrimidine-DNA glycosylase (Fpg). We report the first characterization of a bacterial Ogg, Clostridium acetobutylicum Ogg (CacOgg). Like human OGG1 and Escherichia coli Fpg (EcoFpg), CacOgg excised 8-oxoguanine. However, unlike hOGG1 and EcoFpg, CacOgg showed little preference for the base opposite the damage during base excision and removed 8-oxoguanine from single-stranded DNA. Thus, our results showed unambiguous qualitative functional differences in vitro between CacOgg and both hOGG1 and EcoFpg. CacOgg differs in sequence from the eukaryotic enzymes at two sequence positions, M132 and F179, which align with amino acids (R154 and Y203) in human OGG1 (hOGG1) found to be involved in opposite base interaction. To address the sequence basis for functional differences with respect to opposite base interactions, we prepared three CacOgg variants, M132R, F179Y, and M132R/F179Y. All three variants showed a substantial increase in specificity for 8-oxoG.C relative to 8-oxoG.A. While we were unable to definitively associate these qualitative functional differences with differences in selective pressure between eukaryotes, Clostridia, and other bacteria, our results are consistent with the idea that evolution of Ogg function is based on kinetic control of repair.


Analgesic use and risk of subsequent hypertension in apparently healthy men.
Prospective studies have suggested that women who self-select for use of analgesics have an increased risk of hypertension, but data in men are sparse. We tested whether apparently healthy male physicians who reported analgesic use had an increased risk of subsequent hypertension. Prospective cohort study of 8229 participants in the Physicians' Health Study who were free of hypertension and completed detailed analgesic questionnaires. Hypertension was defined as self-reported blood pressure of 140/90 mm Hg or higher or use of antihypertensive medication. After a mean of 5.8 years' follow-up, 2234 men (27.2%) reported subsequent hypertension. We categorized the cumulative analgesic use in quintiles. After adjusting for potential confounders, men in the highest quintile had no statistically significant increased risk of hypertension (hazard ratio, 1.12; 95% confidence interval, 0.97-1.31) when compared with those in the lowest quintile. In subgroup analyses, we evaluated the cumulative use of nonsteroidal anti-inflammatory drugs, acetaminophen, and aspirin. Compared with never users, men who reported consuming at least 2500 pills had hazard ratios of 1.05 (95% confidence interval, 0.89-1.24) for nonsteroidal anti-inflammatory drugs, 1.08 (95% confidence interval, 0.87-1.34) for acetaminophen, and 1.16 (95% confidence interval, 0.92-1.48) for aspirin. The results were similar for analgesic use in the year preceding the analgesic questionnaire. In this large cohort, apparently healthy male physicians who self-selected for analgesic use had no significantly increased risk of subsequent hypertension, although a small to moderately increased risk cannot be excluded in observational studies.


Myeloid IPMK promotes the resolution of serum transfer-induced arthritis in mice.
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by widespread joint inflammation, which leads to joint damage, disability, and mortality. Among the several types of immune cells, myeloid cells such as macrophages are critical for controlling the pathogenesis of RA. Inositol phosphates are water-soluble signaling molecules, which are synthesized by a series of enzymes including inositol phosphate kinases. Previous studies revealed actions of inositol phosphates and their metabolic enzymes in the modulation of inflammation such as Toll-like receptor-triggered innate immunity. However, the physiological roles of inositol polyphosphate (IP) metabolism in the regulation of RA remain largely uncharacterized. Therefore, our study sought to determine the role of inositol polyphosphate multikinase (IPMK), a key enzyme for IP metabolism and various cellular signaling control mechanisms, in mediating RA. Using myeloid cell-specific IPMK knockout (KO) mice, arthritis was induced via intraperitoneal K/BxN serum injection, after which disease severity was evaluated. Both wild-type and IPMK KO mice developed similar RA phenotypes; however, conditional deletion of IPMK in myeloid cells led to elevated arthritis scores during the resolution phase, suggesting that IPMK deficiency in myeloid cells impairs the resolution of inflammation. Bone marrow-derived IPMK KO macrophages exhibited no apparent defects in immunoglobulin Fc receptor (FcR) activation, osteoclast differentiation, or resolvin signaling. Taken together, our findings suggest that myeloid IPMK is a key determinant of RA resolution.


Molecular structure of mammalian neuropeptide Y: analysis by molecular cloning and computer-aided comparison with crystal structure of avian homologue.
Identification and characterization of the cDNA encoding rat neuropeptide Y revealed the nucleotide sequence coding for a 98-amino acid precursor. The deduced amino acid sequence for rat neuropeptide Y is identical to the human peptide and is highly homologous to avian pancreatic polypeptide. The tertiary structure of avian pancreatic polypeptide has been previously derived from crystallographic data by Blundell and coworkers. The homology between neuropeptide Y and avian pancreatic polypeptide preserves all of the residues essential for the maintenance of the tertiary structure. Thus, it has been possible to compute a three-dimensional model of the mammalian neuropeptide, neuropeptide Y, based on the known structure of the avian homologue. This model suggest that neuropeptide preserves a compact tertiary structure characterized by extensive hydrophobic interactions between an N-terminal polyproline-II-like helix and a C-terminal alpha-helix. The model has been used to identify amino acids residing in key positions within this structure and, thereby, to direct future analysis of neuropeptide Y structure-function relationships.


Peritoneal dialysis solutions low in glucose degradation products: clinical experience and outcomes.
The latest literature describing clinical experiences with peritoneal dialysis solutions low in glucose degradation products (GDPs) is mostly consistent with previous reports suggesting less inflammation, better peritoneal mesothelial mass preservation, a lower rate of decline of residual renal function, and improved patient survival. The data suggest stable peritoneal transport rates, but no definite evidence has yet emerged of superior membrane preservation. Most studies have reported very low peritonitis rates, but without significant differences as compared with rates in patients exposed to conventional solutions. New, appropriately powered randomized clinical trials are needed to confirm the potential benefits of low-GDP solutions and to establish the role of renal function preservation with regard to those benefits.


Drug-targeting methodologies with applications: A review.
Targeted drug delivery to solid tumors is a very active research area, focusing mainly on improved drug formulation and associated best delivery methods/devices. Drug-targeting has the potential to greatly improve drug-delivery efficacy, reduce side effects, and lower the treatment costs. However, the vast majority of drug-targeting studies assume that the drug-particles are already at the target site or at least in its direct vicinity. In this review, drug-delivery methodologies, drug types and drug-delivery devices are discussed with examples in two major application areas: (1) inhaled drug-aerosol delivery into human lung-airways; and (2) intravascular drug-delivery for solid tumor targeting. The major problem addressed is how to deliver efficiently the drug-particles from the entry/infusion point to the target site. So far, most experimental results are based on animal studies. Concerning pulmonary drug delivery, the focus is on the pros and cons of three inhaler types, i.e., pressurized metered dose inhaler, dry powder inhaler and nebulizer, in addition to drug-aerosol formulations. Computational fluid-particle dynamics techniques and the underlying methodology for a smart inhaler system are discussed as well. Concerning intravascular drug-delivery for solid tumor targeting, passive and active targeting are reviewed as well as direct drug-targeting, using optimal delivery of radioactive microspheres to liver tumors as an example. The review concludes with suggestions for future work, considereing both pulmonary drug targeting and direct drug delivery to solid tumors in the vascular system.


Role of TMPRSS4 Modulation in Breast Cancer Cell Proliferation.
Background: TMPRSS4 is a novel Type II transmembrane serine protease found at the surface of the cells and is involved in the development and cancer progression. However, TMPRSS4 functions in breast cancer remain poor understand. The present study investigated the function of TMPRSS4 in the breast cancer cells and the potential mechanistic action underling. Materials and Methods: The lentiviral vectors causing TMPRSS4 down-regulation and over-expression were established and transfected in MDA-MB-468 and MCF-7 cells, respectively. By using the CCK- 8 assay, cell proliferation was analyzed. Moreover, western blot was used to detect the expression of certain proteins related to cell apoptosis (Bax and Bcl2) signaling pathway and telomere maintenance (POT1, TPP1, and UBE2D3). Cell cycle and cell apoptosis were also analyzed by using the Flow cytometry analysis. TMPRSS4 expression was detected at the mRNA level and protein level by performing qPCR and western blot technique, respectively. Results: TMPRSS4 expression is inhibited in stable transfected MDA-MB-468-shTMPRSS4 cells compared to the control MDA-MB-468-NC and its expression is up-regulated in stable transfected MCF-7-TMPTSS4 compared to its control MCF-7-NC. Moreover, TMPRSS4 silencing in breast cancer reduces cells proliferation by promoting cell cycle arrest in G2/M phase, cell apoptosis, and telomere maintenance impairment while the TMPRSS4 overexpression increases cells proliferation through cell apoptosis reduction and telomere maintenance reinforcement associated with insignificant change in cell cycle progression. Conclusion: TMPRSS4 plays important roles in cancer progression and may be considered as a good therapeutic target for cancer gene therapy especially breast cancer.


Impact of an acute pain service on pain outcomes with combat-injured soldiers at Camp Bastion, Afghanistan.
Few studies demonstrate the impact of early aggressive analgesia with acute pain service (APS) involvement at combat support hospitals (CSHs) using real-time data. Collaboration between the British and the United States (US) Army led to a 3-month deployment of a US Army APS to Camp Bastion, the main British military base in southern Afghanistan, from April to July 2009. Pain outcomes data were collected at Camp Bastion from 71 soldiers sustaining major combat injuries followed by an APS. The sample was predominantly male (98.6%) and Caucasian (98.8%) with a mean age 25.4 years ± 5.4 (range 18-45). Regional nerve blocks were performed in 51 (71.8%) and epidurals placed in 11 (15.5%) of the cases. Repeated measures analysis of variance showed a statistically significant decreases in pain intensity (numeric rating scale from 0 for "none" to 10 for "as bad as you can imagine") over the three data collection points; first 3, 4-6, and 7-10 hours (F = 133.35; degrees of freedom [df; 1,68]; P < 0.001). Mean percent pain relief (0% "no relief" to 100% "complete relief") increased significantly over time (F = 193.12; df[1,69]; P < 0.001) with scores 31.29% ± 20.3 (initial 3 hours), 74.86% ± 27.5 (4-6 hours), and 83.14% ± 19.3 (7-24 hours). Overall, the perceived quality of pain control by soldiers was high. Findings from this quality of pain care study show that aggressive multimodal analgesia interventions by an APS in a CSH is associated with decreased pain intensity and increased pain relief.


Effects of zopiclone and benzodiazepine hypnotics on search in short-term memory.
The effects of zopiclone 7.5 mg in comparison to flunitrazepam 1 mg, triazolam 0.25 mg and lormetazepam 1 mg on stages of information processing in a memory-scanning task were investigated in 10 normal volunteers using a double-blind placebo-controlled cross-over design. Overall reaction times for the test were significantly increased 1 h following treatment with zopiclone, flunitrazepam and triazolam and in the morning following nighttime medication with flunitrazepam. The effects of zopiclone, flunitrazepam and triazolam were found to be located in the serial comparison and response-selection organization stages of the reaction process although both flunitrazepam and triazolam caused additional impairment of stimulus encoding.


Adolescent Motivation Toward Physical Exercise: The Role of Sex, Age, Enjoyment, and Anxiety.
Physical exercise declines during adolescence due to several factors, such as lack of intrinsic motivation or enjoyment, high physical exercise anxiety, and so on. The objectives of this study were to validate the psychometric structure of the Behavioral Regulation in Exercise Questionnaire-2 to confirm the existence of five levels of motivation toward physical exercise in adolescent population, as well as to analyze the differences in motivation according to sex and age, and the role of age and sex in the relationship between motivation and enjoyment. To do so, 666 students between 10 and 16 years old completed the Behavioral Regulation in Exercise Questionnaire-2 and questionnaires related to enjoyment and physical exercise anxiety. The confirmatory factor analysis of the Behavioral Regulation in Exercise Questionnaire-2 showed a five-factor structure. However, a more parsimonious four-factor structure with a single intrinsic-identified regulation emerged through an exploratory factor analysis. The more self-determined types of motivation were positively associated with enjoyment and negatively with anxiety, the type of physical exercise motivation fluctuated depending on age and sex, and the age moderated the relationship between motivation and enjoyment. This study highlights the importance of fostering specific types of motivation to improve the physical exercise and the relevance of age and sex when developing interventions.


[Efficacy of anal dilators in the treatment of acute anal fissure. A controlled clinical trial].
Procedures involving the use of anal dilators or topical nitroderivatives for the treatment of anal fissure are efficacious, economic and safe. The aim of our study was to compare the efficacy of two conservative treatments - passive dilation with anal dilators or topical nitroderivatives - in reducing anal pressure and resolving anal fissures. A total of 40 patients with a clinical diagnosis of acute anal fissure in the absence of hypotonic anal sphincter, abscess or perianal fistula, haemorrhoidal thrombosis, chronic inflammatory bowel disease or lower gastrointestinal neoplasia were randomly assigned to treatment with dilators (20 patients) or topical nitroderivatives (20 patients). After 4 weeks of treatment, 90% of patients treated with dilators and only 45% treated with topical nitroderivatives showed complete resolution of their anal fissures and a reduction of sphincter hypertone. After 12 weeks, 2/18 patients successfully treated with dilators and 1/9 patients successfully treated with topical derivatives presented recurrence of the anal fissure. The use of anal dilators would appear to induce better resolution of acute anal fissures than topical nitroderivates, as confirmed by the low relapse rate at 12 weeks.


Relationship between Trunk Muscle Mass Index at Admission and Walking Independence in Patients with Hip Fracture.
This study aimed to investigate the relationship between trunk muscle mass index (TMI), appendicular skeletal muscle mass index (ASMI), and walking independence in patients aged 65 years and older undergoing rehabilitation for hip fracture. This retrospective, observational study was conducted in a convalescent rehabilitation ward and included 314 patients (aged ≥65 years) with hip fracture. The patients were classified into the independence group [functional independence measure (FIM)-walk score ≥6] or the non-independence group (FIM-walk score ≤5) according to the mobility item score among the motor FIM items at the time of discharge. Age, sex, TMI, ASMI, and Mini Nutritional Assessment-Short Form (MNA-SF) data were also extracted. Between-group and multivariate analyses were performed to evaluate the factors associated with walking independence. The independence group had higher TMI (males: 6.6±0.9 vs. 5.6±1.0 kg/m2, P <0.001; females: 6.1±0.8 vs. 5.7±1.0 kg/m2, P <0.001), ASMI (males: 6.7±1.1 vs. 5.9±1.3 kg/m2, P=0.004; females: 5.3±0.9 vs. 4.7±0.8 kg/m2, P <0.001), MMSE-J (21.5±4.9 vs. 16.4±4.5 points, P <0.001), and MNA-SF [median (interquartile range): 8 (6-9) vs. 7 (5-8) points, P <0.001] than the non-independence group. Multivariate analysis showed that TMI at admission was significantly associated with walking independence (odds ratio: 1.86, 95% confidence interval: 1.28-2.72, P <0.001). This study suggests that a higher TMI at admission was important for acquiring walking independence in patients with hip fracture and shows the importance of early evaluation of TMI during hospitalization of patients with hip fracture.


S14G-Humanin ameliorates ovalbumin-induced airway inflammation in asthma mediated by inhibition of toll-like receptor 4 (TLR4) expression and the nuclear factor κ-B (NF-κB)/early growth response protein-1 (Egr-1) pathway.
Asthma is a chronic inflammatory disease with a high morbidity rate in children and significantly impacts their healthy growth. It is reported that Th2 cell-mediated airway inflammation and activated oxidative stress are involved in the pathogenesis of asthma. S14G-humanin (HNG) is a derivative of Humanin with higher activity. The present study proposes to explore the potential treating property of HNG on asthma. An asthma model was constructed in mice using ovalbumin (OVA), the mice were treated with 2.5 mg/kg and 5 mg/kg HNG for 16 days. Dramatically increased lung weight index, elevated number of monocytes, eosinophils, and neutrophils, promoted production of Th2 cytokines including interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL-13), and severe histological pathology were observed in OVA-challenged mice, all of which were extremely alleviated by 2.5 mg/kg and 5 mg/kg HNG. Furthermore, the increased malondialdehyde (MDA) level and declined superoxide dismutase (SOD) activity in OVA-challenged mice were abolished by 2.5 mg/kg and 5 mg/kg HNG. Lastly, the upregulated TLR4, p-NF-κB p65, and early growth response 1 (Egr-1) in lung tissues of OVA-challenged mice were pronouncedly downregulated by 2.5 mg/kg and 5 mg/kg HNG. Collectively, our data suggested that HNG ameliorated airway inflammation in asthma partially due to NF-κB and Egr-1-mediated responses.


Minimal Excitations in the Fractional Quantum Hall Regime.
We study the minimal excitations of fractional quantum Hall edges, extending the notion of levitons to interacting systems. Using both perturbative and exact calculations, we show that they arise in response to a Lorentzian potential with quantized flux. They carry an integer charge, thus involving several Laughlin quasiparticles, and leave a Poissonian signature in a Hanbury Brown-Twiss partition noise measurement at low transparency. This makes them readily accessible experimentally, ultimately offering the opportunity to study real-time transport of Abelian and non-Abelian excitations.


Biodistribution of nanostructured lipid carriers: a tomographic study.
This study describes the preparation, characterization, and biodistribution of radiolabelled nanostructured lipid carriers (NLCs) especially designed for in vivo tomographic study. A preliminary formulative study was conducted in order to incorporate (99m)Tc based tracer in NLCs. At this aim a (99m)Tc complex containing a terminal (99m)Tc ≡ N multiple bond ([(99m)Tc]N-DBODC2) has been synthesized and included in NLCs produced by a stirring and ultrasonication method. The morphological and dimensional characteristics of the produced NLCs have been accurately investigated by a number of specific techniques, including: cryogenic transmission electron microscopy, X-ray, photon correlation spectroscopy and sedimentation field flow fractionation. The obtained NLCs were employed for achieving in vivo tomographic images of the rat body by small-animal SPECT scanner that enabled the investigation of NLC biodistribution after intraperitoneal, intravenous, intranasal and oral administration. NLC production protocol allowed to firmly encapsulate the radiotracer within the nanoparticles. In vivo studies evidenced that NLC remained stable in vivo, suggesting their suitability as controlled release system for drugs and radiochemical for therapeutic and diagnostic purposes. Moreover the high resolution images obtained by the SPECT technique allowed to detect NLC presence in brown fat tissue, suggesting NLC therapeutic application for treating human obesity and related metabolic disorders.


Identification method of raindrops and hailstones based on digital holographic interference.
The identification of raindrops and hailstones is of great significance to the study of precipitation characteristics from the aspect of microphysics and can provide important data support for weather modification. In this paper, an identification method of raindrops and hailstones based on digital holographic interference is proposed. The grayscale gradient variance method is used to obtain the focus position of the particles. By means of binarization and morphological processing, digital holograms are processed to obtain clear profiles of the particles. Then the contour parameters of the particles are used to obtain the equivalent volume diameter and roundness. Finally, according to the equivalent volume diameter, roundness and lens-like effect of the particles, the phase states of the raindrop and hailstone are identified by the algorithm. Experiments show that the method proposed in this paper has a good identification effect on raindrops and hailstones. The research results can provide reference for the research of the identification method of raindrops and hailstones and the acquisition of accurate characteristic parameters.


Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition.
The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67-antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly "irregular" endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67-antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.


Combination therapy in hypertension.
There is controversy as to which BP-lowering manoeuvres and/or drugs can be combined effectively. Ideally, a combination therapy should show an additional BP-lowering effect over monotherapy. The effect should be of sufficient clinical significance, without short- or long-term adverse effects, and the patients should feel well. Combination therapy should be based on the understanding of the mechanisms of action of the different classes of drugs used and of their interaction. Rational combinations include an ACE inhibitor or a beta-blocker together with moderate salt restriction or a thiazide diuretic or a calcium antagonist. Less rational combinations include an ACE inhibitor with a beta-blocker, and a calcium antagonist with a thiazide diuretic.


MicroRNAs: control and loss of control in human physiology and disease.
Analysis of the human genome indicates that a large fraction of the genome sequences are RNAs that do not encode any proteins, also known as non-coding RNAs. MicroRNAs (miRNAs) are a group of small non-coding RNA molecules 20-22 nucleotides (nt) in length that are predicted to control the activity of approximately 30% of all protein-coding genes in mammals. miRNAs play important roles in many diseases, including cancer, cardiovascular disease, and immune disorders. The expression of miRNAs can be regulated by epigenetic modification, DNA copy number change, and genetic mutations. miRNAs can serve as a valuable therapeutic target for a large number of diseases. For miRNAs with oncogenic capabilities, potential therapies include miRNA silencing, antisense blocking, and miRNA modifications. For miRNAs with tumor suppression functions, overexpression of those miRNAs might be a useful strategy to inhibit tumor growth. In this review, we discuss the current progress of miRNA research, regulation of miRNA expression, prediction of miRNA targets, and regulatory role of miRNAs in human physiology and diseases, with a specific focus on miRNAs in pancreatic cancer, liver cancer, colorectal cancer, cardiovascular disease, the immune system, and infectious disease. This review provides valuable information for clinicians and researchers who want to recognize the newest advances in this new field and identify possible lines of investigation in miRNAs as important mediators in human physiology and diseases.


Different Angiotensin-converting enzyme inhibitors and the associations with overall and cause-specific mortalities in patients with hypertension.
Angiotensin-converting enzyme (ACE) inhibitors have been widely used in the treatment of hypertension, but the comparative effectiveness in reducing mortality among different drugs is seldom reported. We identified hypertensive patients who started captopril, enalapril, lisinopril, fosinopril, perindopril, ramipril, or imidapril therapy from Taiwan's National Health Insurance database between 1 January 2004 and 31 December 2009. Overall and cause-specific mortalities were ascertained through a linkage to Taiwan's National Death Registry. Patients were followed from the initiation of ACE inhibitors to death, disenrollment, or study termination (31 December 2010). A Cox proportional hazard regression model was used to calculate the hazard ratio (HR) and 95% confidence interval (CI), using ramipril as the reference group. A total of 989,489 hypertensive patients were included, with a mean follow-up ranging from 3.5 years for imidapril to 4.5 years for enalapril. Captopril initiators had the highest overall mortality rate (117.8 per 1,000,000 person-days) as compared to other ACE inhibitors (54.3-79.4 per 1,000,000 person-days). Patients who started captopril therapy had a significantly increased risk of overall mortality (HR: 1.28, 95% CI: 1.24-1.31) when compared with ramipril. Enalapril (HR: 1.08, 95% CI: 1.05-1.11) and fosinopril (HR: 1.08, 95% CI: 1.05-1.12) were also associated with a modestly increased risk. No difference in mortality was found for lisinopril, perindopril, and imidapril, as compared with ramipril. There are differences in the mortality risk associated with different ACE inhibitors. However, potential residual confounding effects might still exist.


Claudin-18 splice variant 2 is a pan-cancer target suitable for therapeutic antibody development.
Antibody-based cancer therapies have emerged as the most promising therapeutics in oncology. The purpose of this study was to discover novel targets for therapeutic antibodies in solid cancer. We combined data mining and wet-bench experiments to identify strictly gastrocyte lineage-specific cell surface molecules and to validate them as therapeutic antibody targets. We identified isoform 2 of the tight junction molecule claudin-18 (CLDN18.2) as a highly selective cell lineage marker. Its expression in normal tissues is strictly confined to differentiated epithelial cells of the gastric mucosa, but it is absent from the gastric stem cell zone. CLDN18.2 is retained on malignant transformation and is expressed in a significant proportion of primary gastric cancers and the metastases thereof. In addition to its orthotopic expression, we found frequent ectopic activation of CLDN18.2 in pancreatic, esophageal, ovarian, and lung tumors, correlating with distinct histologic subtypes. The activation of CLDN18.2 depends on the binding of the transcription factor cyclic AMP-responsive element binding protein to its unmethylated consensus site. Most importantly, we were able to raise monoclonal antibodies that bind to CLDN18.2 but not to its lung-specific splice variant and recognize the antigen on the surface of cancer cells. Its highly restricted expression pattern in normal tissues, its frequent ectopic activation in a diversity of human cancers, and the ability to specifically target this molecule at the cell surface of tumor cells qualify CLDN18.2 as a novel, highly attractive pan-cancer target for the antibody therapy of epithelial tumors.


Operable malignant jaundice: To stent or not to stent before the operation?
Traditionally, pre-operative biliary drainage (PBD) was believed to improve multi-organ dysfunction, and for this reason, was practiced worldwide. Over the last decade, this concept was challenged by many reports, including meta-analyses that showed no difference in morbidity and mortality between surgery with, and surgery without PBD, in operable malignant jaundice. The main disadvantages of PBD are seen to be the additional cost of the procedure itself, and the need for longer hospitalization. In addition, many studies showed the significance of specific complications resulting from PBD, such as recurrent jaundice, cholangitis, pancreatitis, cutaneous fistula, and bleeding. However, the results of these studies remain inconclusive as to date there has been no perfect study that equally randomized comparable patients according to the level of obstruction and technique used for PBD. Generally, endoscopic stent insertion (ES) is preferred for common duct obstruction, whereas endoscopic nasobiliary drainage and percutaneous biliary drainage is reserved for hilar obstruction, since ES in hilar block confers a high rate of cholangitis. Although, there is no guideline which either supports or refutes this approach, certain subgroups of patients, including those with symptomatic jaundice, cholangitis, impending renal failure, hilar block requiring preoperative portal vein embolization, and those who need pre-operative neoadjuvant therapy, are suitable candidates for PBD.


LncRNAs elevate plant adaptation under low temperature by maintaining local chromatin landscape.
Epigenetic regulation is one of the most precise and subtle ways of gene regulation, including DNA modification, histone modification, RNA modification, histone variants, chromatin remodeling, and long non-coding RNAs (lncRNAs). Chromatin modification is the most basic type of epigenetic regulation, which plays a key role in a myriad of developmental and physiological processes that have been thoroughly studied. These modifications are usually completed by a series of conserved chromatin modification complexes in eukaryotes. In recent years, a series of lncRNAs in organisms also have been described as having irreplaceable functions in biological environment adaptation, especially in biotic and abiotic stresses. Moreover, these molecules form a sophisticated regulatory network through mutual cross-regulation to achieve quantitative expression of key environmental response genes to external signals. For instance, the function of lncRNAs will directly or indirectly depend on the function of the chromatin modification complex. In this review, we mainly focus on chromatin modification, lncRNA, and their coordination mechanism to achieve the high adaptability of plants in low-temperature environments. We highlight recent findings and insights into lncRNA-mediated local chromatin environment changes during plant growth under low temperature via chromatin modification complexes, including target gene specificity for different lncRNA.


Chronic inflammatory changes in the pouch mucosa are associated with cholangitis found on peroperative liver biopsy specimens at restorative proctocolectomy for ulcerative colitis.
The clinical syndrome of primary sclerosing cholangitis (PSC), diagnosed in about 5% of patients with ulcerative colitis (UC), has been shown to be associated with pouchitis after ileal pouch-anal anastomosis. The aim of this study was to ascertain whether UC patients with cholangitis on liver biopsy at proctocolectomy, with or without the clinical syndrome of PSC, have an increased risk of inflammatory changes in the ileal reservoir mucosa and clinical pouchitis. Of the consecutive 81 UC patients treated with restorative proctocolectomy with ileal J reservoir at Tampere University Hospital between 1985 and 1991, 73 with peroperative liver biopsy were included. A peroperative liver biopsy was obtained during proctocolectomy. After a median follow up of 64 months, pouch biopsy specimens were obtained. Periods of clinical pouchitis were diagnosed by means of clinical criteria alone or by clinical criteria combined with the results of previous pouch endoscopies in all patients. Ten patients (14%) showed histologic features consistent with small-duct PSC on liver biopsy. Endoscopic retrograde cholangiography had previously been performed on four of these patients, and all four had large-duct PSC. Patients with cholangitis had significantly more severe chronic, but not acute, inflammation in the pouch mucosa than patients without cholangitis. At least one episode of pouchitis occurred in 30% of the patients without cholangitis as compared with 90% of the patients with cholangitis. Chronic pouchitis was more frequent in the group with cholangitis than in the group without it (70% versus 11%). The only means of detecting all UC patients with cholangitis is a liver biopsy. Cholangitis, either with the clinical syndrome of PSC or found on liver biopsy, seems to be a risk factor for chronic-type inflammatory changes in the pouch mucosa and for the development of pouchitis.


A review on architecture of the gag-pol ribosomal frameshifting RNA in human immunodeficiency virus: a variability survey of virus genotypes.
Programmed '-1' ribosomal frameshifting is necessary for expressing the pol gene overlapped from a gag of human immunodeficiency virus. A viral RNA structure that requires base pairing across the overlapping sequence region suggests a mechanism of regulating ribosome and helicase traffic during expression. To get precise roles of an element around the frameshift site, a review on architecture of the frameshifting RNA is performed in combination of reported information with augments of a representative set of 19 viral samples. In spite of a different length for the viral RNAs, a canonical comparison on the element sequence allocation is performed for viewing variability associations between virus genotypes. Additionally, recent and historical insights recognized in frameshifting regulation are looked back as for indel and single nucleotide polymorphism of RNA. As specially noted, structural changes at a frameshift site, the spacer sequence, and a three-helix junction element, as well as two Watson-Crick base pairs near a bulge and a C-G pair close a loop, are the most vital strategies for the virus frameshifting regulations. All of structural changes, which are dependent upon specific sequence variations, facilitate an elucidation about the RNA element conformation-dependent mechanism for frameshifting. These facts on disrupting base pair interactions also allow solving the problem of competition between ribosome and helicase on a same RNA template, common to single-stranded RNA viruses. In a broad perspective, each new insight of frameshifting regulation in the competition systems introduced by the RNA element construct changes will offer a compelling target for antiviral therapy.


[The middle-forearm flap based on perforator of ulnar artery for electrical burn wound on the wrist ].
To dicuss the application and therapeutic effect of middle-forearm flap based on perforator of ulnar artery for electrical burn wound on the wrist. From Oct. 2009 to Oct. 2012, 10 cases of electrical burn wounds on the wrist were treated. A line from radialis medial epicondyle of humerus to the interior radialis pisiform bone was connected as flap axis. At the midpoint of the line, Doppler flow imaging meter was used to detect the emerging point of perforator vessel. The flap was designed and harvested. The flap was transferred reversely, with superficial vein retaining which was anastomosed with vein at recipient sites in 3 cases. The wounds in the donor sites were closed directly in 2 cases, and with skin graft in 8 cases. All the 10 flaps survived completely. 7 cases without vein anastomosis underwent obvious flap edema during 2-4 days postoperatively, which resovled 1 week later. Sub-flap tissue necrosis and infection happened in 2 cases, which healed after dressing and drainage. Patients were followed up for 3-36 months with satisfactory results. The middle-forearm flap based on perforator of ulnar artery has a stable and reliable blood supply. It offers a new choice for the electric burn wound on the wrist, especially at the ulnar side.


Top-down and bottom-up interaction: manipulating the dichotic listening ear advantage.
The aim of the study was to investigate interactions between top-down and bottom-up information processing in the auditory domain. For this purpose, thirty-five right-handed participants with normal hearing acuity were tested with consonant-vowel dichotic stimulus pairs. Bottom-up stimulus characteristics were manipulated by gradually varying interaural intensity difference from -21 dB in favor of the left ear to +21 dB in favor of the right ear (including a no difference baseline condition). Top-down manipulation consisted of three conditions with different attention instructions: one free report condition, and each one condition requiring the participants to focus their attention on the right ear and on the left ear, respectively. The results showed a significant interaction of bottom-up and top-down manipulations with respect to the modulation of the ear advantage. Post-hoc analysis showed that the effect of directing attention was reduced when the intensity difference favored the to-be-attended ear. Thus, bottom-up intensity and top-down attention manipulations should not be regarded as independent but rather interacting factors when it comes to the manipulation of the ear advantage in a dichotic listening situation.


Effect of the moisture content in aerosol on the spray performance of Stmerin ® D hydrofluoroalkane preparations (2).
The spray performance of the new Stmerin® D pressurized metered dose inhalers (MDIs), prescribed for asthma treatment, was previously reported to be sensitive to moisture content in the canister. In this formulation, active ingredients were suspended in hydrofluoroalkanes (HFA-134a and HFA-227). Here, we report on the critical moisture content of a new MDI formulation using HFA-227. We prepared MDIs with different moisture contents (107-331 ppm) and investigated the effect of moisture content on the fine particle dose (FPD). When the moisture content exceeded 280 ppm, FPD was reduced by 20%. The moisture content can be kept below this level by controlling moisture ingress with selected sealing gasket materials and by controlling the manufacturing conditions. The long-term stability and moisture penetration rate at 25°C and 60% relative humidity (RH) of the new HFA-227 formulation were tested over 36 months. Moisture content increased to approximately 210 ppm after 24 months and 230 ppm after 36 months storage; the drug content did not change over 24 months; FPD was slightly reduced by 24 months but still complied with the product specification. These results demonstrate that the critical moisture content of the Stmerin® D HFA-227 MDI formulation to meet required spray performance is between 280 and 330 ppm, and this formulation (at <210 ppm moisture level) is stable over 24 months.


Peptide backbone conformation affects the substrate preference of protein arginine methyltransferase I.
Asymmetric dimethylation of arginine side chains is a common post-translational modification of eukaryotic proteins, which serves mostly to regulate protein-protein interactions. The modification is catalyzed by type I protein arginine methyltransferases, PRMT1 being the predominant member of the family. Determinants of substrate specificity of these enzymes are poorly understood. The Nuclear poly(A) binding protein 1 (PABPN1) is methylated by PRMT1 at 13 arginine residues located in RXR sequences in the protein's C-terminal domain. We have identified a preferred site for PRMT1-catalyzed methylation in PABPN1 and in a corresponding synthetic peptide. Variants of these substrates were analyzed by steady-state kinetic analysis and mass spectrometry. The data indicate that initial methylation is directed toward the preferred arginine residue by an N-terminally adjacent proline. Enhanced methylation upon peptide cyclization suggests that induction of a reverse turn structure is the basis for the ability of the respective proline residue to enable preferred methylation of the neighboring arginine residue, and this notion is supported by far-UV circular dichroism spectroscopy. We suggest that the formation of a reverse turn facilitates the access of arginine side chains to the active sites of PRMT1, which are located in the central cavity of a doughnut-shaped PRMT1 homodimer.


Generation of Two Biological Wound Dressings as a Potential Delivery System of Human Adipose-Derived Mesenchymal Stem Cells.
Human adipose-derived mesenchymal stem cells (hADMSCs) are believed to be potential key factors for starting the regenerative process after tissue injury. However, an efficient method of delivering these regenerative cells to an external wound site is still lacking. Human amnion and pig skin have long been used as skin wound dressings for the treatment of burns and other skin lesions. Herein, we present the generation of two constructs using these two biomaterials as effective scaffolds for the culture of hADMSCs. It was found that hADMSCs seeded onto radiosterilized human amnion and pig skin are viable and proliferate. These cells are able to migrate over these scaffolds as demonstrated by using time-lapse microscopy. In addition, the scaffolds induce hADMSCs to secrete interleukin-10, an important negative regulator of inflammation, and interleukin-1β, a proinflammatory protein. The interplay between these two proteins has been proven to be vital for a balanced restoration of all necessary tissues. Thus, radiosterilized human amnion and pig skin are likely suitable scaffolds for delivery of hADMSCs transplants that could promote tissue regeneration in skin injuries like patients with burn injuries.


Infrequent detection of cytomegalovirus and Epstein-Barr virus DNA in synovial membrane of patients with rheumatoid arthritis.
To study the role of the cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus types 1 and 2 (HSV-1 and 2), varicella zoster virus (VZV), and human herpes virus 6 (HHV-6) in the etiology of rheumatoid arthritis (RA). Polymerase chain reaction (PCR) was used to detect DNA of the different herpes viruses in synovial membranes from 31 patients with chronic RA and 14 control patients. Specific antibodies were determined by indirect immunofluorescence and ELISA. Out of 31 patients with RA, CMV DNA was detected in synovial membranes from 2 patients and EBV DNA was detected in synovial membranes from 2 other patients. All samples from the patients with RA were negative for DNA from HSV-1 and 2, VZV, and HHV-6. All samples from the 14 control patients were negative in all PCR assays. No statistically significant differences in IgG antibodies were found for CMV, HSV-1, VZV, and HHV-6 in patients with RA compared to controls. Higher titers of IgG antibodies against EBV viral capsid antigen were found in patients with RA, with a significance of p < 0.05. Both CMV and EBV DNA were detected in synovial membranes from 6% of the patients with RA. We cannot exclude the possibility that these viruses were associated with disease development in a minority of patients with RA.


Microglia and neurodegeneration: the role of systemic inflammation.
It is well accepted that CNS inflammation has a role in the progression of chronic neurodegenerative disease, although the mechanisms through which this occurs are still unclear. The inflammatory response during most chronic neurodegenerative disease is dominated by the microglia and mechanisms by which these cells contribute to neuronal damage and degeneration are the subject of intense study. More recently it has emerged that systemic inflammation has a significant role to play in the progression of these diseases. Well-described adaptive pathways exist to transduce systemic inflammatory signals to the brain, but activation of these pathways appears to be deleterious to the brain if the acute insult is sufficiently robust, as in severe sepsis, or sufficiently prolonged, as in repeated stimulation with robust doses of inflammogens such as lipopolysaccharide (LPS). Significantly, moderate doses of inflammogens produce new pathology in the brain and exacerbate or accelerate features of disease when superimposed upon existing pathology or in the context of genetic predisposition. It is now apparent in multiple chronic disease states, and in ageing, that microglia are primed by prior pathology, or by genetic predisposition, to respond more vigorously to subsequent inflammatory stimulation, thus transforming an adaptive CNS inflammatory response to systemic inflammation, into one that has deleterious consequences for the individual. In this review, the preclinical and clinical evidence supporting a significant role for systemic inflammation in chronic neurodegenerative diseases will be discussed. Mechanisms by which microglia might effect neuronal damage and dysfunction, as a consequence of systemic stimulation, will be highlighted.


28-day repeated dose toxicity and toxicokinetics study on new melatonergic antidepressant GW117 in beagle dogs.
GW117 is new melatonergic antidepressant being developed to show better antidepressant action than agomelatine. The purpose of this study was to evaluate the toxicity and to determine potential target organs after oral (gavage) administration of the test article GW117 for 28 days and to assess the reversibility after a 4-week recovery phase in beagle dogs. Toxicokinetics was also evaluated. Four groups were designed in this study, including the vehicle control group and the GW117 50, 150 and 500 mg/kg/day groups, with 5 dogs/sex/group. Body weight, hematology, clinical chemistry, gross necropsy, organ weight, histopathology, and other indicators were examined. Results showed that animals dosed at ≥150 mg/kg/day showed gastrointestinal reactions (watery feces and dark green/red brown feces), with a dose-response relationship in the incidence and severity grade. Female dogs at 500 mg/kg/day had an increase in organ weight and ratios of the liver at the end of the dosing phase. Histopathology examination showed that some animals at 500 mg/kg/day, especially female animals, had minimal centrilobular hepatocyte hypertrophy in the liver, which reversed after 28-day recovery. With the exception of the above, no GW117-related abnormality was noted. Meanwhile, there were no sexual differences in drug exposure and accumulation after the first and last dosing. The no observed adverse effect dose level (NOAEL) was 150 mg/kg/day, under which mean Cmax and AUC0 → t were 583.5 and 2767.0 ng/ml*h for females and 663.2 and 4046.3 ng/ml*h for males on Day 28.


Congenital heart defects in CTNNB1 syndrome: Raising clinical awareness.
CTNNB1 [OMIM *116806] encodes β-catenin, an integral part of the cadherin/catenin complex, which functions as effector of Wnt signaling. CTNNB1 is highly expressed in brain as well as in other tissues, including heart. Heterozygous CTNNB1 pathogenic variations are associated with a neurodevelopmental disorder characterized by spastic diplegia and visual defects (NEDSDV) [OMIM #615075], featuring psychomotor delay, intellectual disability, behavioral disturbances, movement disorders, visual defects and subtle facial and somatic features. We report on a new series of 19 NEDSDV patients (mean age 10.3 years), nine of whom bearing novel CTNNB1 variants. Notably, five patients showed congenital heart anomalies including absent pulmonary valve with intact ventricular septum, atrioventricular canal with hypoplastic aortic arch, tetralogy of Fallot, and mitral valve prolapse. We focused on the cardiac phenotype characterizing such cases and reviewed the congenital heart defects in previously reported NEDSDV patients. While congenital heart defects had occasionally been reported so far, the present findings configure a higher rate of cardiac anomalies, suggesting dedicated heart examination to NEDSDV clinical management.


A 2D Coordination Network That Detects Nitro Explosives in Water, Catalyzes Baylis-Hillman Reactions, and Undergoes Unusual 2D→3D Single-Crystal to Single-Crystal Transformation.
The solvothermal reaction of Zn(NO3)2·6H2O and a linear dicarboxylate ligand H2L, in the presence of urotropine in N,N'-dimethylformamide (DMF), gives rise to a new porous two-dimensional (2D) coordination network, {[Zn3(L)3(urotropine)2]·2DMF·3H2O}n (1), with hxl topology. Interestingly, framework 1 exhibits excellent emission properties owing to the presence of naphthalene moiety in the linker H2L, that can be efficiently suppressed by subtle quantity of nitro explosives in aqueous medium. Furthermore, presence of urotropine molecules bound to the metal centers, 1 is found to be an excellent heterogeneous catalyst meant for atom-economical C-C bond-forming Baylis-Hillman reactions. Additionally, crystals of 1 undergo complete transmetalation with Cu(II) to afford isostructural 1Cu. Moreover, the 2D framework of 1 allows replacement of urotropine molecules by 4,4'-azopyridine (azp) linker resulting in a three-dimensional (3D) metal-organic framework, {[Zn(L)(azp)]·4DMF 2H2O}n (2). The 1→2 transformation takes place in single-crystal-to-single crystal manner supported by powder X-ray diffraction, atomic force microscopy, high-resolution transmission electron microscopy, and morphological studies. Remarkably, during this 2D→3D transformation, the original trinuclear [Zn3(COO)6] secondary building unit changes to a mononuclear node, which is unprecedented.


The inhibitory effects of bioactive compounds of tomato juice binding to hepatic HMGCR: in vivo study and molecular modelling.
The hypocholesterolemic effect of tomato juice has been investigated in an intervention study with rats, along with the possible inhibition effect of bioactive tomato compounds binding to the HMGCR enzyme. Two experimental groups (n = 8 Sprague-Dawley rats) were fed ad libitum for five weeks, with water or tomato juice provided to the control and intervention groups, respectively. Total, LDL and HDL cholesterol, and total triglycerides were analysed in plasma, and the lycopene content and the expression and activity of the enzyme HMGCR were determined in liver samples. A computational molecular modelling was carried out to determine the interactions between HMGCR and lycopene, chlorogenic acid and naringenin. Total, LDL and HDL cholesterol were significantly lower in the intervention group after the intake of tomato juice. In addition, a significant reduction in HMGCR activity was observed, although this was not accompanied by changes in gene expression. The molecular modelling showed that components of tomato can bind to the active site of the enzyme and compete with the ligand HMGCoA. Lycopene, from tomato juice, accumulates in the liver and can inhibit the activity of the rate-limiting enzyme of cholesterol biosynthesis, HMGCR.


Increased PPARD Expression May Play a Protective Role in Human Lung Adenocarcinoma and Squamous Cell Carcinoma.
Peroxisome proliferator-activated receptor-δ, encoded by gene PPARD, is overexpressed in a majority of human lung cancer subtypes, but its role in the tumor progression remains poorly understood. We have analyzed the expression of PPARD in lung adenocarcinoma (LA) and squamous cell carcinoma (LSCC) datasets. The potential roles of PPARD in the pathological development of LA and LSCC were explored through literature-based pathway analysis and pathway enrichment analysis. In all LA datasets (N = 11) and in seven out of nine LSCC studies, the levels of PPARD were increased as compared to control tissues (log-fold changes were 0.37 ± 0.20 and 0.10 ± 0.37 for LA and LSCC, respectively). On average, the expression levels of PPARD in LA were higher than those in LSCC (p = 0.036). Pathway analysis showed that the overexpression of PPARD might play both positive and negative roles in the development of both LA and LSCC. Specifically, PPARD inhibits seven LSCC promoters and seven LA promoters and activates one LSCC inhibitor and another LA inhibitor. However, PPARD also activates six and one promoters of LA and LSCC, respectively, which would facilitate the development of LA/LSCC. Our results suggested a mixed role of PPARD in LA/LSCC, which may add new insights into the understanding of the PPARD-lung cancer relationship.


Partial purification and characterization of the reverse transcriptase of the simian immunodeficiency virus TYO-7 isolated from an African green monkey.
The reverse transcriptase (RT) was partially purified by a newly developed procedure from the simian immunodeficiency virus TYO-7 isolated from an African green monkey (SIVagmTYO-7). The method comprised lysis of the virus with nonionic detergent followed by two centrifugations in isopycnic sucrose density gradients and one velocity sedimentation in a glycerol gradient. The enzyme exhibited a purity of 70-80% and showed an exceptional high specific activity of 135 nmol incorporation of dTMP per milligram of protein in 1 h with poly(rA).oligo(dT) as template-primer (TP). The molecular weight of the native enzyme was estimated by velocity sedimentation analysis as 120K-130K. Investigation of the RT by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the active enzyme is a heterodimer composed of a 64- and a 50-kDa subunit. The two subunits were identified to be RT specific by Western blot analysis. In activity gels, both subunits exhibited enzymatic activity, whereby the 64-kDa subunit showed the predominant activity. The RT preferred the TP poly(rA).oligo(dT) over poly(rC).oligo(dG). With poly(rCm).oligo(dG), only marginal activity was detected, and no activity was measured with poly(dA).oligo(dT). The TP specificity was influenced by the reaction temperature. The highest activity was measured around the melting temperature of the TP used. Furthermore, the enzyme activity was more thermolabile when measured with poly(rA).oligo(dT) than with poly(rC).oligo(dG). To compare the specificity of RT inhibitors, their inhibition efficiency (IE) was defined as the ratio of the 50% inhibiting concentration (ID50) obtained with the RT in viral lysates to the ID50 of purified RT.


Structural brain network topological alterations in stuttering adults.
Persistent developmental stuttering is a speech disorder that primarily affects normal speech fluency but encompasses a complex set of symptoms ranging from reduced sensorimotor integration to socioemotional challenges. Here, we investigated the whole-brain structural connectome and its topological alterations in adults who stutter. Diffusion-weighted imaging data of 33 subjects (13 adults who stutter and 20 fluent speakers) were obtained along with a stuttering severity evaluation. The structural brain network properties were analysed using network-based statistics and graph theoretical measures particularly focussing on community structure, network hubs and controllability. Bayesian power estimation was used to assess the reliability of the structural connectivity differences by examining the effect size. The analysis revealed reliable and wide-spread decreases in connectivity for adults who stutter in regions associated with sensorimotor, cognitive, emotional and memory-related functions. The community detection algorithms revealed different subnetworks for fluent speakers and adults who stutter, indicating considerable network adaptation in adults who stutter. Average and modal controllability differed between groups in a subnetwork encompassing frontal brain regions and parts of the basal ganglia. The results revealed extensive structural network alterations and substantial adaptation in neural architecture in adults who stutter well beyond the sensorimotor network. These findings highlight the impact of the neurodevelopmental effects of persistent stuttering on neural organization and the importance of examining the full structural connectome and the network alterations that underscore the behavioural phenotype.


A Silicon Sub-Bandgap Near-Infrared Photodetector with High Detectivity Based on Textured Si/Au Nanoparticle Schottky Junctions Covered with Graphene Film.
We present a straightforward approach to develop a high-detectivity silicon (Si) sub-bandgap near-infrared (NIR) photodetector (PD) based on textured Si/Au nanoparticle (NP) Schottky junctions coated with graphene film. This is a photovoltaic-type PD that operates at 0 V bias. The texturing of Si is to trap light for NIR absorption enhancement, and Schottky junctions facilitate sub-bandgap NIR absorption and internal photoemission. Both Au NPs and the texturing of Si were made in self-organized processes. Graphene offers additional pathways for hot electron transport and to increase photocurrent. Under 1319 nm illumination at room temperature, a responsivity of 3.9 mA/W and detectivity of 7.2 × 1010 cm × (Hz)1/2/W were obtained. Additionally, at -60 °C, the detectivity increased to 1.5 × 1011 cm × (Hz)1/2/W, with the dark current density reduced and responsivity unchanged. The result of this work demonstrates a facile method to create high-performance Si sub-bandgap NIR PDs for promising applications at ambient temperatures.


Effect of titanium surface texture on the cell-biomaterial interface.
To improve the in vivo performance of engineered implants, this study examines the independent effects of surface chemistry and topography on fibroblast morphology and density in vitro. Titanium (Ti) was sputter-coated onto smooth and microtextured polyethylene terephthalate (PET). Test specimens were evaluated in 24-h, fibroblast cultures and assessed with scanning electron microscopy (SEM) for cellular morphology and density. Fibroblast density increased, as the Ti film thickness increased. The fibroblasts exhibited contact guidance on the textured test specimens. The greatest cellular density was found on the Ti-coated, textured test specimens. In conclusion, Ti and surface texture appeared to strongly influence fibroblast density and morphology as compared to PET and smooth surfaces.


Rapid and sensitive method for the determination of sertraline in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
A method for the determination of sertraline, a new antidepressant drug and a selective serotonin reuptake inhibitor (SSRI), in human plasma is described. Therapeutic drug monitoring (TDM) necessitates efficient, fast and reliable analytical methods validated by external quality control. We therefore devised a simple, rapid and sensitive isocratic reversed-phase liquid chromatographic-tandem mass spectrometric method equipped with Turbo Ion spray (TIS) source, operating in the positive ion and selective reaction monitoring (SRM) acquisition mode to quantify sertraline in human plasma. A new and superior procedure of solid-phase extraction (SPE) (compared to liquid-liquid extraction) was followed to extract sertraline and imipramine as internal standard (IS) from the human plasma. Sample preparation was performed using waters hydrophilic-lipophilic balance (HLB) cartridge and this method yielded extremely clean extracts with very good recovery, 81.47 and 85.79% for sertraline and IS, respectively. Both were analyzed by combined reverse phase liquid chromatography and tandem mass spectrometry (LC-MS/MS) with positive ion TIS ionization using SRM acquisition mode. The response of the LC-MS/MS method for sertraline was linear over the dynamic range of 0.5-60.0 ng/ml with correlation coefficient r>or=0.9996. The coefficient of variance (%CV) was 8.53% at 0.5 ng/ml and the accuracy was well within the accepted limit of +/-20% at lower limit of quantification (LLOQ) and +/-15% at all the other concentrations in the linear range. This method was fully validated for the accuracy, precision and stability studies. The above findings indicate that the method is very much accurate and precise and can be successfully applied for bioequivalence studies in human subjects.


Treatment of Recalcitrant Nonunion of the Radius With Posterior Interosseous Bone Flap in Children.
Forearm nonunion is a rare condition in children, and its treatment is usually successful. However, nontraumatic nonunion can be resistant to treatment. Although the vascularized fibular bone graft has been frequently used for complex forearm nonunion, there is no consensus on the best treatment method. In this study, we aimed to investigate the outcome of the antegrade posterior interosseous bone flap (PIBF) in treating children with complex radius nonunion. The participants consisted of 3 girls and 1 boy ranging from 31 months to 9 years of age. They were treated with PIBF surgery, and the nonunion of the radius was fixed with a plate or an external fixator. The nonunion was due to congenital pseudoarthrosis, osteogenesis imperfecta, or infantile osteomyelitis. All patients were evaluated for a clinical and radiologic union. The radiologic union was achieved 2 to 3 months after the surgery in all cases. The functional recovery of the elbow, forearm, and wrist was satisfactory except for 1 patient who experienced donor site fracture, and finally radioulnar synostosis accompanied by radial head dislocation. Because of the simplicity of its technique and short duration of surgery, PIBF can be a reliable and less-demanding alternative to vascularized fibular bone graft in the treatment of complex forearm nonunion in children. Level IV.


Significance of CASP8AP2 and H2AFZ expression in survival and risk of relapse in children with acute lymphoblastic leukemia.
Novel biomarkers for risk refinement and stratification in childhood acute lymphoblastic leukemia (ALL) are needed to optimize treatment results. We studied the expression of CASP8AP2 and H2AFZ associated with relapse and survival in bone marrow samples from newly diagnosed children with ALL. We found: (a) an increased risk for early relapse in those patients with low expression of CASP8AP2 (odds ratio [OR] 3.93, 95% confidence interval [CI] 1.40-11.02, p < 0.05) confirming its usefulness as a predictive risk marker, although H2AFZ did not present the same effect; (b) patients with low expressions of CASP8AP2 and H2AFZ had inferior survival rates (p < 0.001); (c) the predictive values regarding low expressions of H2AFZ and CASP8AP2 and high white blood cell count suggest that these features could help to identify more accurately patients at greater risk of relapse.


Hard- and Soft-Tissue Reconstruction in the Anterior Maxilla for Esthetic Implant Therapy.
Implant rehabilitation in the esthetic zone is often challenged by vertical bone defects and soft-tissue deformities. This article describes a combined hard- and soft-tissue restorative approach that involves staged guided bone regeneration, implant placement, and two soft-tissue augmentation procedures to achieve optimal esthetic outcomes at multiple implant sites in the anterior zone. The staged bone augmentation procedure, performed with a mixture of autogenous and xenogeneic bone graft and a nonresorbable membrane, allowed for the placement of three implants in ideal positions after 9 months. Further soft-tissue augmentation involved the use of multiple connective tissue grafts (CTGs) stabilized on the occlusal aspect of the implants and between the implants to enhance peri-implant papillae (ie, the "iceberg" CTG approach). Then, a second soft-tissue grafting procedure was executed to reposition the mucogingival junction and re-establish an adequate amount of keratinized mucosa at the implant sites. The article highlights the importance of performing both hard- and soft-tissue augmentation for implant therapy in the esthetic zone.


[The COP-BLAM therapy for non-Hodgkin's lymphoma in elderly patients].
COP-BLAM therapy, which has recently been reported to be useful in the treatment of malignant lymphoma, was performed on aged patients with non-Hodgkin's lymphoma, and the results and adverse effects of the treatment were evaluated. The subjects of the present study included 13 patients with non-Hodgkin's lymphoma, aged 60 years or older, treated during the period from October, 1987 to June, 1989. They included four recurrent cases after CHOP therapy. According to the Ann-Arbor classification of pathological stage there were two patients in stage I, one in stage II, two stage III and eight in stage IV. COP-BLAM therapy was conducted using the following procedure, irrespective of the patient's age, intravenous drip infusion of cyclophosphamide (CPM) at a dose of 400 mg/m2 and intravenous infusion of 1 mg/m2 vincristine (VCR), and 30 mg/m2 adriamycin (ADM), on day 1; oral administration of 40 mg/m2 prednisone (PSL), and 100 mg/m2 procarbazine (PCZ), from day 1 through day 10, followed by intravenous infusion of bleomycin (BLM), 10 mg/body on day 14. BLM was withdrawn or decreased to less than 70% of the initial dose in patients who exhibited disturbed pulmonary function. Treatment resulted in complete remission (CR) in 11 and partial remission (PR) in 2 out of the 13 patients. Analyzing results according to pathological stage. CR was obtained in all of those in staged I and II, and eight out of the 10 in staged III and IV. The four patients with recurrent disease after CHOP therapy exhibited a CR rate of 50%, indicating that therapy is highly effective in recurrent cases.(ABSTRACT TRUNCATED AT 250 WORDS)


Exposure to cotton dust in an experimental cardroom.
Changes in respiratory function (FEV1) and polymorphonuclear neutrophils (PMN) on nasal epithelium were studied in 68 students and 39 cotton mill workers in an experimental cardroom. The exposure was characterised by the vertical elutriator dust and endotoxin levels. A dose related decrease was found for FEV1 which was more pronounced in smoking workers. The thresholds for no FEV1 reaction were 0.58 mg/m3 dust and 0.17 micrograms/m3 endotoxin for students and 0.43 mg/m3 and 0.08 micrograms/m3 for smoking workers. The PMN increased in most of the experiments but no dose response relationship could be shown. In experiments with smoking workers or when washed cotton was carded a better correlation was obtained between FEV1 decrease and endotoxin levels than for dust levels.


A perfectly matched layer for fluid-solid problems: Application to ocean-acoustics simulations with solid ocean bottoms.
A time-domain Legendre spectral-element method is described for full-wave simulation of ocean acoustics models, i.e., coupled fluid-solid problems in unbounded or semi-infinite domains, taking into account shear wave propagation in the ocean bottom. The technique can accommodate range-dependent and depth-dependent wave speed and density, as well as steep ocean floor topography. For truncation of the infinite domain, to efficiently absorb outgoing waves, a fluid-solid complex-frequency-shifted unsplit perfectly matched layer is introduced based on the complex coordinate stretching technique. The complex stretching is rigorously taken into account in the derivation of the fluid-solid matching condition inside the absorbing layer, which has never been done before in the time domain. Two implementations are designed: a convolutional formulation and an auxiliary differential equation formulation because the latter allows for implementation of high-order time schemes, leading to reduced numerical dispersion and dissipation, a topic of importance, in particular, in long-range ocean acoustics simulations. The method is validated for a two dimensional fluid-solid Pekeris waveguide and for a three dimensional seamount model, which shows that the technique is accurate and numerically long-time stable. Compared with widely used paraxial absorbing boundary conditions, the perfectly matched layer is significantly more efficient at absorbing both body waves and interface waves.


Cervicoplasty: nonexcisional anterior approach.
The purpose of this paper is to demonstrate a technique of cevicoplasty by means of a submental incision exclusively, without removal of skin. This technique is indicated in patients with good-quality skin that has adequate capacity for contraction and redraping. The neck is approached by means of a 3- to 4-cm incision posterior to the submental crease. A thick subcutaneous flap is developed. Initial "open" dissection is continued with dissection using the endoscope. All the fat over the platysma muscle is dissected off. The dissection follows a triangle outlined by the sternocleidomastoid muscles. An insulated suction coagulator is used for hemostasis. Conservative defatting of the subcutaneous layer is done, and no suction lipectomy is used routinely. However, all the fat pad between the platysma borders and the fat pad between the mylohyoid and the digastric muscles are removed. The digastric muscles are either "shaved off" or plicated in the midline with 3-0 nylon sutures. This provides a smooth and flat contour to the submental area. Subplatysmal dissection beyond the submaxillary salivary gland is done with the aid of the endoscope. After a trial of advancement, the medial borders of the platysma are resected, and plication in the midline is performed from the level of the thyroid cartilage to the symphysis of the mandible. In most cases, a backcut on the platysma from the thyroid cartilage level in an oblique direction for about 3 to 4 cm is done to avoid secondary lateral platysma bands. If no good definition on the submental angle is obtained, an interlocked suture suspension with anchoring to the mastoid fascia is performed. The relative value of this maneuver will be discussed. The skin is allowed to redrape. Usually, after medial advancement of the platysma, an additional platysma-skin separation is needed in some segments to allow smooth contour on the surface of the skin. The suture suspension may leave minimal rippling laterally. This usually disappears in a few days. The technique described has been used for several years as an isolated procedure or as a part of a full endoscopic face lift without skin excisions. If used in the right patient, the result is comparable with that of the open excisional approach and has a high rate of satisfaction. The long-term durability remains to be seen.