Variability and repeatability of noctule bat migration in Central Europe: evidence for partial and differential migration.
Each year, large numbers of bats move across Europe between their summer and winter areas, yet even though many of them are endangered and legally protected, we are unaware about many aspects of their migratory behaviour. Here, taking Nyctalus noctula as a model species, we used stable hydrogen isotopic values in fur ( δ2Hf) as an endogenous marker to shed light on the migratory behaviour of more than 1000 bats from hibernacula across Central Europe. Specifically, we asked the following questions: how flexible is migration in temperate zone bats? Which general migration pattern do noctule bats follow? How repeatable and thus predictable is the migratory behaviour of individuals? Do morphological correlates of migration occur in bats? Our study confirmed that noctule bats engage in partial and female-biased migration across Europe, suggesting the strongest migration pressures for northern populations. Further, we revealed a combination of partial and differential migration patterns with highly variable migration distances which lead to a pronounced mixing of different source populations in hibernacula where mating occurs. Most individuals were consistent in their migration strategy over time, i.e. 86% could be repeatedly assigned to either long-distance or regional origin across years. This is consistent with our finding that the between-individual component explained 84% of the variation in δ2Hf values, suggesting specialized individual migratory behaviours and a strong natal philopatry. We discovered a positive correlation between forearm length and migration distance and support for sex-specific effects of migration on body condition. Our study elucidated migration patterns over large geographical scales, demonstrating that considerable numbers of migratory bats originating from distant populations depend on hibernacula across Central Europe, calling for international conservation management.


The health thinkers: clarifying health determinants but not upsetting sinners.
Probably no health thinker has played a greater part than Professor Tom McKeown in bringing medical practice in modern society under close and critical scrutiny. He has accomplished this not by a ruthless assessment of individual clinical therapies, but by linking the role of medicine to the wider issues within epidemiology.


Klingsor syndrome: A rare surgical emergency.
Klingsor syndrome or self-Inflicted traumatic penile amputation is a rare clinical entity that is associated with psychiatric disorders, command hallucinations, religious preoccupations, substance abuse, and isolation from or neglect by society. In addition to being infrequently encountered, it is a rare surgical emergency, with paucity of data on appropriate and timely management to ensure optimal functional outcomes. We report the case of a 25-year-old unmarried male who inflicted this injury upon himself in a fit of paranoia as a way to expiate his sins and earn solace from God. An attempt at microvascular re-implantation 12 h after the injury was successful, with adequate cosmetic and functional outcomes obtained. Thus, in this rare disease, despite a delay in presentation, a single attempt at re-implantation may still be worthwhile for obtaining optimal cosmetic and psychosocial benefits.


Ultra-Early Cerebral Thrombosis Formation After Experimental Subarachnoid Hemorrhage Detected on T2* Magnetic Resonance Imaging.
The mechanisms of brain damage during ultra-early subarachnoid hemorrhage (SAH) have not been well studied. The current study examined the SAH-induced hyperacute brain damage at 4 hours using magnetic resonance imaging and brain histology in a mouse model. SAH was induced by endovascular perforation in adult mice. First, adult male wild-type mice underwent magnetic resonance imaging T2 and T2* 4 hours after an endovascular perforation or a sham operation and were euthanized to assess brain histology. Second, male and female adult lipocalin-2 knockout mice had SAH. All animals underwent magnetic resonance imaging at 4 hours, and the brains were harvested for brain histology. T2* hypointensity vessels were observed in the brain 4 hours after SAH in male wild-type mice. The numbers of T2*-positive vessels were significantly higher in SAH brains than in sham-operated mice. Brain histology showed thrombosis and erythrocyte plugs in the T2*-positive cerebral vessels which may be venules. The number of T2*-positive vessels correlated with SAH grade and the presence of T2 lesions. Brain thrombosis was also accompanied by albumin leakage and neuronal injury. LCN2 deficient male mice had lower numbers of T2*-positive vessels after SAH compared with wild-type male mice. SAH causes ultra-early brain vessel thrombosis that can be detected by T2* gradient-echo sequence at 4 hours after SAH. LCN2 deficiency decreased the number of T2*-positive vessels.


Intensive granulocyte and monocyte adsorption apheresis for generalized pustular psoriasis.
Granulocyte and monocyte adsorption apheresis (GMA) is usually performed weekly (consisting of five sessions) for refractory skin diseases, such as generalized pustular psoriasis (GPP). The time to remission of inflammatory bowel diseases has been reported to be significantly shorter in intensive GMA (twice a week) than in regular GMA (once a week). Despite several reports of GPP cases treated with intensive GMA, the efficacy of intensive GMA has not been verified in GPP. Herein, we present two GPP patients with a mutation in the IL36RN gene, who initially received regular GMA, and intensive GMA upon recurrence. There were no adverse effects during regular and intensive GMA for both patients. Because concomitant medication was only prednisolone (20 mg/day) during regular and intensive GMA, intensive GMA showed superiority to regular GMA in patient 1. Although concomitant medications were different between regular and intensive GMA in patient 2, these drugs had been used before the start of each GMA therapy. We cannot neglect the effects of concomitant drugs, but we observed a shorter time to remission in intensive GMA than that in regular GMA in both patients. More case studies will be necessary for evaluating the clinical efficacy of intensive GMA.


[Histomorphological studies on hyperplastic prostate of castrated rat caused by androgen].
To investigate prostatic histomorphological changes of rat benign prostatic hyperplasia (BPH) models. After castrated, SD rats were injected subcutaneously testosterone propionate to induce the BPH. Water substitution method was used to measure prostatic volumes. Prostatic tissue were stained by hemoloxylin and eosin, the morphometric changes of glandular and interstitial tissues were semi-quantified by image analysis system. Multiple linear regression was adopted to analyze the results. In comparison with normal group, prostatic volumes were significantly enlarged (P < 0.01) with glands expanded and interstitial tissues increased in BPH model. Glandular average diameters, volumes and surface areas in unit volume, as well as glandular circumferences and glandular relative total volumes were all significantly increased (P < 0.01). Glandular counts, density, ratio of glandular surface area to volume. And glandular average curvature were all declined (P < 0.05-0.01), so was interstitial circumference (P < 0.01). But volume density had no changes, and relative total interstitial volumes were obviously increased. Prostatic volume was significantly correlated with glandular relative total volume (r = 0.989, P < 0.001) and interstitial relative total volume(r = 0.789, P < 0.001). Prostatic volume was also correlated significantly with glandular average volume(r = 0.789, P < 0.001). Action of androgen on rat prostate may mainly lie in glandular epithelial hyperplasia, which manifests enlargement of glandular lumen accompanied by hyperplasia of interstitial tissues.


Breast Milk for Term and Preterm Infants-Own Mother's Milk or Donor Milk?
Hormones are important biological regulators, controlling development and physiological processes throughout life. We investigated pituitary hormones such as follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and total protein levels during the first 6 months of lactation. Breast milk samples were collected every fourth week of lactation from mothers who gave birth to preterm (n = 14) or term (n = 16) infants. Donor milk is suggested when own mother's milk is not available; therefore, we collected breast milk samples before and after Holder pasteurization (HoP) from the Breast Milk Collection Center of Pécs, Hungary. Three infant formulas prepared in the Neonatal Intensive Care Unit of the University of Pécs were tested at three different time points. Our aim was to examine the hormone content of own mother's milk and donor milk. There were no significant changes over time in the concentrations of any hormone. Preterm milk had higher PRL (28.2 ± 2.5 vs 19.3 ± 2.3 ng/mL) and LH (36.3 ± 8.8 vs 15.9 ± 4.1 mIU/L) concentrations than term milk during the first 6 months of lactation. Total protein and FSH concentrations did not differ between preterm and term breast milk. Holder pasteurization decreased the PRL concentration (30.4 ± 1.8 vs 14.4 ± 0.6 ng/mL) and did not affect gonadotropin levels of donor milk. Infant formulas have higher total protein content than breast milk but do not contain detectable levels of pituitary hormones. Differences were detected in the content of pituitary hormones produced for preterm and term infants. Divergence between feeding options offers opportunities for improvement of nutritional guidelines for both hospital and home feeding practices.


Placebo-controlled trial of cisapride in postoperative ileus.
The effect of 4 mg of the gastrointestinal motility enhancing agent cisapride on postoperative ileus was compared with that of a placebo in 53 patients who had undergone various types of surgery. They received one i.v. injection of the double-blind medication, or two if no distinct flatus occurred within an hour after the first injection. The placebo response was very limited; borborygmi or flatus did not occur in more than 12% of the patients not even after the second injection. Cisapride was significantly effective: bowel sounds were present in 43% and 50% within 1 hour after the first and second injection of 4 mg, respectively, and flatus in 36% and 43%. The recognition that colon inactivity probably is the major determinant of the ileus, may be indicative of a substantial relative impact of cisapride's stimulating effects on colon motility.


Quaternary Solar Cells with 12.5% Efficiency Enabled with Non-Fullerene and Fullerene Acceptor Guests to Improve Open Circuit Voltage and Film Morphology.
Designing the bulk-heterojunction structure (binary, ternary and quaternary) is of great fundamental interest for relaxing the trade-off between open circuit voltage (Voc ) and short circuit current density ( Jsc ). Herein, a new quaternary blended material system is reported with a nonfullerene and PC71 BM as the third and fourth component of PBDB-T:ITCT, leading to simultaneously increased Voc and Jsc and maintained fill factor (FF). The guest IT-T-IC has upshifted LUMO energy level helping to obtain a higher Voc . The further addition of 0.2 PC71 BM as the fourth component yields an even higher Voc because the LUMO of PC71 BM is higher than that of ITCT. Interestingly, the blend of PC71 BM leads to the formation of an unprecedented neuron-like morphology, which acts as new centers not only performing light absorption and charge separation but also charge transport through their surrounding donor and acceptor fibers. The increased hole and electron mobilities and the reduced bimolecular loss results in an even larger Jsc and FF. These results indicate that a combination using a structurally similar higher-LUMO-level non-fullerene acceptor and PCBM is a simple yet effective quaternary material approach to simultaneously increase Voc and Jsc while maintaining FF, improving final device performance.


[Experience of the use of silodosin in acute urinary retention caused by benign prostatic hyperplasia].
For the treatment of acute urinary retention (AUR) as one of the most serious complications of adenoma of the prostate (BPH), alpha-adrenoblockers are widely used. The article presents an experience of the use of the new uroselective alpha-adrenoblocker silodosin approved for the treatment of patients with urination disorders caused by BPH. Its pharmacological profile has a number of advantages, including the highest uroselectiveness at the present day, immediate action, the potential for the use of standard dose of 8 mg 1 time a day, which does not require a correction depending on the age, and the possibility of the simultaneous application with antihypertensive drugs.


Strain dependency of the transfer of experimental immune insulitis in mice.
Multiple treatment with low doses of streptozotocin induces hyperglycaemia with concomitant lymphocytic infiltrations into pancreatic islets (insulitis) in several mouse strains. The transfer of cellular immune reactions against islet cells by means of spleen cells was tested in two congeneic and five allogeneic strain combinations. Donor mice were treated on 5 consecutive days with 40 mg streptozotocin per kg body weight. Three weeks later, 5 x 10(7) live spleen cells were transferred into thymusless recipient mice. Insulitis which had developed in about 70% of the donors was only transferable from C57B1/6J to congeneic thymusless mice. In a second congeneic and in all allogeneic strain combinations, cellular immune reactions against pancreatic islets could not be transferred. In none of the recipients of spleen cells from diabetic donors was hyperglycaemia observed. As streptozotocin-induced cellular immune reactions against pancreatic islet cells were only transferable in one congeneic and in no allogeneic strain combinations, it is concluded that there is a genetic restriction both on the levels of donor and recipient mice.


Comparative aspects of actions of a short-chain phospholipid on epithelial Na+ channels and tight junction conductance.
Ion transport in both the frog skin (a high-resistance epithelium) and the rabbit nasal airway epithelium (a low-resistance epithelium) are dominated by electrogenic Na+ absorption via apical membrane amiloride-sensitive Na+ channels, and short-circuit current (ISC) is essentially a measure of Na+ absorption in both epithelia. In both epithelia, mucosal application of the short-chain phospholipid didecanoyl-L-alpha-phosphatidylcholine (DDPC) dose-dependently inhibited the amiloride-sensitive ISC and caused an initial decrease in epithelial conductance (Gt) followed by an increase in Gt to steady-state values above control level. The effects were reversible. It is concluded that DDPC (a) inhibits epithelial amiloride-sensitive Na+ channels and (b) induces an increase in paracellular tight junction conductance. These effects may involve changes in non-specific lipid-protein interactions at the cell membrane level.


Localization of Ca2+ extrusion sites in pancreatic acinar cells.
We have investigated the localization of Ca2+ extrusion sites in mouse pancreatic acinar cells. Employing a new technique, in which high resolution localization of cellular Ca2+ exit is achieved by confocal microscopy and a Ca2+-sensitive fluorescent probe coupled to heavy dextran to slow down diffusion of extracellular Ca2+, it is shown directly that the secretory pole (secretory granule area) is the major site for Ca2+ extrusion following agonist stimulation. This Ca2+ extrusion appears not to be a consequence of exocytosis, as assessment of secretion under our experimental conditions (low external Ca2+ concentration, room temperature) using the technique of monitoring quinacrine fluorescence shows little loss of secretory granules in spite of sustained Ca2+ exit. We conclude that Ca2+ is primarily extruded by Ca2+ pumps from the secretory pole and propose that this process is useful for maintaining a high Ca2+ concentration in the acinar lumen, which is necessary for promotion of endocytosis.


Methyl (Z)-2-{[N-(2-formyl-phen-yl)-4-methyl-benzene-sulfonamido]-meth-yl}-3-phenyl-prop-2-enoate.
In the title compound, C(25)H(23)NO(5)S, the sulfonyl-bound benzene ring forms dihedral angles of 37.2 (1) and 67.0 (1)°, respectively, with the formyl-phenyl and phenyl rings. The mol-ecular conformation is stabilized by an intra-molecular C-H⋯π inter-action. In the crystal, mol-ecules are linked by C-H⋯O hydrogen bonds, forming a two-dimensional network in the (110) plane in which R(4) (4)(38) ring motifs are generated.


Propofol-remifentanil-based anaesthesia vs. sevoflurane-fentanyl-based anaesthesia for immediate postoperative ophthalmic evaluation following strabismus surgery.
Following strabismus surgery, immediate postoperative ophthalmic evaluation may be desired. Thus, an anaesthetic technique allowing rapid recovery of ocular motility is required. Saccadic eye movements is a biophysical monitor of ocular motility and may be used to assess recovery from anaesthesia. The aim of this study is to compare the time to the recovery of saccadic eye movements in patients, following one of two anaesthetic techniques: Propofol-remifentanil-based anaesthesia vs. sevoflurane-fentanyl-based anaesthesia. Fifty adult patients undergoing strabismus surgery were randomly assigned to one of two groups: patients in Group R received induction and maintenance of anaesthesia with propofol and remifentanil, while patients in Group S received induction of anaesthesia with propofol and fentanyl and maintenance of anaesthesia with sevoflurane. Recovery from anaesthesia was measured from the time all anaesthetics were turned off and was assessed every 2 min. Recovery time was attained when patients were able to generate brisk saccadic eye movements. At recovery time, the ophthalmic evaluation was started. The mean recovery time of saccadic eye movements was significantly shorter in the Group R when compared to the Group S (12.1 +/- 4.3 min vs. 21.5 +/- 4.7 min, respectively, P < 0.0001). More patients in Group S experienced nausea and vomiting postoperatively as compared to Group R (9/25 vs. 2/25, respectively, P = 0.037). Propofol-remifentanil-based anaesthesia may be a useful technique in strabismus surgery when immediate postoperative ophthalmic evaluation is desired. When compared to sevoflurane maintenance of anaesthesia, it allows for a more rapid recovery from anaesthesia as judged by recovery of saccadic eye movements and a decreased incidence of postoperative nausea and vomiting.


Dominant negative activity of mutant thyroid hormone alpha1 receptors from patients with hepatocellular carcinoma.
Complementary DNAs for two mutant thyroid hormone alpha1 receptors (TR alpha1) were isolated from hepatocellular carcinomas of two patients. Sequence analyses of the complementary DNAs showed a single Val390Ala and double Pro398Ser/Glu350Lys mutations in mutants H and L, respectively. We characterized their hormone-binding, DNA-binding, and dominant negative activities. Mutants H and L did not bind the hormone T3. Their DNA-binding activities were analyzed using three types of thyroid hormone response elements (TREs) in which the half-site binding motifs are arranged in an everted repeat (Lys), an inverted repeat (Pal), or a direct repeat separated by four nucleotides (DR4). Compared with wild-type TR alpha1 (w-TR alpha1), which bound these TREs with different homodimer/monomer ratios, binding of mutant L to the three TREs as homodimers was reduced by approximately 90%. However, binding of mutant H to these TREs was more complex. Although it bound normally to DR4 as homodimers, its binding to Lys as homodimers was reduced by approximately 80%. Surprisingly, its binding to Pal was markedly enhanced compared with w-TR alpha1. The binding of these two mutants to the three TREs as heterodimers with retinoid X receptors (RXR alpha and -beta) was not significantly affected. Consistent with the lack of T3-binding activity, both mutants had lost their trans-activation capacity. Mutants H and L exhibited dominant negative activity, but differed in their TRE dependency. The dominant negative potency of mutant H was in the rank order of Pal > DR4 > Lys, whereas no TRE dependency was observed for mutant L. The present study indicates that mutations of the TR alpha gene do occur in patients and that these novel TR alpha1 mutants provide a valuable tool to further understand the molecular basis of the dominant negative action of mutant TRs.


Alkyl Silica Hybrid Nanowire Assembly in Improved Superhydrophobic Membranes for RO Filtration.
Alkyl silica membranes and wires were synthesized by a sol-gel method, which has the capacity to control the size of the particles or membranes by controlling the reactions. Trimethoxyoctylsilane (C8TMOS) was used as a chemical surfactant; poly(vinylpyrrolidone) (PVP) as an emulsifier, dissolved in butanol for emulsion; and tetraethylorthosilicate (TEOS) as a precursor and a source of silica. An assembly of silica wires was fabricated on glass and cotton substrates by the dip-coating technique. Porous membranes and silica wires were observed using scanning electron microscopy (SEM) images. The contact angles of all of the samples were in the range of 140-154° as measured by ImageJ software, which confirmed the hydrophobic nature of the samples. The contact angle was increased by increasing the amount of the surfactant. Phase changes of silica wires and membranes were investigated by thermogravimetric analysis. Chemical bonds of the sample were studied using Fourier transform infrared (FTIR) spectroscopy. The band gap of silica nanowires was measured to be 3.8-3.4 eV using the UV-visible spectrum and decreased as compared to that of bulk silica. These silica-based porous membranes with enhanced transport properties can be used in filtration and separation techniques. This fabricated hybrid silica membrane showed ∼96% salt rejection within a permeation flux of 3.04 L/m2 h.


[Effect of estradiol benzoate on the heat shock protein 70, 90 expressions in endometrial glandular epithelial cell].
To explore the effect of estradiol benzoate on the expressions of heat shock protein (HSP) 70, 90 in endometrial glandular epithelial cell. Normal endometrial glandular epithelial cells were isolated, and cultured by enzymolysis method and identified by electron microscopy and immunohistochemical analysis. The normal endometrial glandular epithelial cells were treated with culture medium only, estradiol benzoate (10(-9), 10(-8), 10(-7) or 10(-6) mol/L), estradiol benzoate (10(-9), 10(-8), 10(-7) or 10(-6) mol/L) and antiestrogen ICI 182780 (fulvestrant, faslodex, 1 micromol/L) and ICI 182780 only for 6, 12, 18, 24 hours respectively. The dose- and time-related effect of estradiol benzoate and ICI 182780 on the cell growth was measured by mononuclear cell direct cytotoxicity assay (methyl thiazolyl tetrazolium assay), and that on the expression of HSP70 and HSP90 in normal endometrial glandular epithelial cell in vitro was measured by immunohistochemical analysis and computerized image analysis system. Estradiol benzoate stimulated cell growth in a time- and dose-dependent manner and the effect was attenuated by the antiestrogen ICI 182780. The average cell growth rates of 10(-9), 10(-8), 10(-7), 10(-6) mol/L estradiol benzoate for 24 hours were (170 +/- 9)%, (207 +/- 11)%, (231 +/- 12)%, (257 +/- 10)%, which were significantly higher than those of 6 hours [(117 +/- 13)%, (129 +/- 10)%, (146 +/- 10)%, (176 +/- 6)%, P < 0.05] and that those estradiol + ICI 182780 for 24 hours [(137 +/- 4)%, (145 +/- 10)%, (151 +/- 9)%, (167 +/- 3)%, P < 0.05]. Estradiol benzoate treatment resulted in a time- and dose-dependent increase of the expression of HSP90 and decrease of HSP70 in human endometrial glandular epithelial cell. The expression of HSP90 in 10(-9), 10(-8), 10(-7), 10(-6) mol/L estradiol benzoate for 24 hours were (64.8 +/- 10.7)%, (75.9 +/- 12.6)%, (80.4 +/- 8.2)%, (83.2 +/- 7.6)%, which were significantly higher than that of the control [(28.0 +/- 3.3)%, (29.0 +/- 5.6)%, (29.0 +/- 5.0)%, (30.0 +/- 6.4)%, P < 0.05] and that of estradiol benzoate + ICI 182780 for 24 hours [(28.2 +/- 2.1)%, (29.7 +/- 3.2)%, (35.0 +/- 4.7)%, (34.7 +/- 6.5)%, P < 0.05]. The expression of HSP70 in 10(-9), 10(-8), 10(-7), 10(-6) mol/L estradiol benzoate for 24 hours were (38.4 +/- 5.7)%, (35.3 +/- 4.9)%, (32.5 +/- 3.1)%, (25.3 +/- 6.2)%, which were significantly lower than that of the control. The estradiol benzoate-induced effects on HSP70 levels were not attenuated by the antiestrogen ICI 182780. The effects of estradiol benzoate on the cell growth and HSP90 expression are estrogen receptor-dependent and the expression of HSP90 may be beneficial to the cell growth. The effect of estradiol on the expression of HSP70 is estrogen receptor-independent and the expression of HSP70 may protect the cell from damage.


A difficult to diagnose case of low-grade endometrial stromal sarcoma with smooth muscle differentiation treated with laparoscopic surgery: A case report.
Low-grade endometrial stromal sarcoma (LGESS) is a rare uterine tumor, accounting for <1% of all uterine cancer cases. LGESS has several variations and the frequency of tumors exhibiting smooth muscle differentiation is 10-30% of LGESS cases, making such cases even rarer. The present report described the case of a patient with LGESS with smooth muscle differentiation, who was diagnosed as having uterine leiomyoma by preoperative needle biopsy and then underwent laparoscopic surgery. The patient was a 41-year-old woman. MRI findings revealed a diffusely hyperintense uterine tumor on T2-weighted images, thus needle biopsy was performed. This tumor was initially diagnosed as leiomyoma, due to the pathological findings of the biopsied specimen, which possessed tumor cells with spindle-shaped nuclei arranged in a cord and positive immunostaining for smooth muscle actin. The patient was subsequently followed up, and MRI findings after 29 months showed tumor growth. Needle biopsy was performed again and the findings were the same as those of the first biopsy; therefore, this tumor was diagnosed as a leiomyoma and laparoscopic hysterectomy was performed. However, the pathological findings of the excised uterus showed small round tumor cells and CD10 immunostaining positivity, thus the tumor was finally diagnosed as LGESS. The patient requested to be followed up and has shown no signs of recurrence 20 months after surgery. The results of retrospective examination in this case suggested that the presence of regions where only CD10 was positive in immunostaining analysis for SMA and CD10 was useful for needle biopsy diagnosis of LGESS with smooth muscle differentiation.


Distribution and evaluation of bone and soft tissue tumors operated in a tertiary care center.
The aim of this study was to retrospectively evaluate the patients who were operated in our orthopedics and traumatology clinic with the suspection of bone and soft tissue tumors. A total of 3133 patients (1146 (46.5%) female and 1318 (53.5%) male) who presented to our tertiary clinic from different regions of Turkey between January 2002 and July 2013 with the presumed diagnosis of bone and soft tissue tumors were analyzed according to age, gender, bone/soft tissue localization, tumoral localization, histopathological diagnosis, tumor size and incidence. Of all operated patients, 2464 (78%) were diagnosed with tumor, while non-tumoral causes were found in 669 (22%) patients. Of the cases diagnosed with tumor, 1139 were bone localized, 1004 soft tissue localized, and 321 metastasis. The most common benign bone tumors were osteochondroma (130, 20%), enchondroma (96, 15%), and simple bone cysts (90, 14%), while the most common malignant bone tumors were osteosarcoma (241, 44%), ewing's sarcoma (89, 16%), and chondrosarcoma (77, 14%); respectively. The most common benign soft tissue tumors were lipoma (141, 22%), giant cell tumors (108, 16%) and ganglion (107, 16%), while the most common malignant soft tissue tumors were liposarcoma (55, 16%), synovial sarcoma (53, 16%) and malignant mesenchymal tumors (45, 13%); respectively. Musculoskeletal tumors are rare, but descriptive data in any region are important in order to reduce mortality and improve treatment. No significant difference was found between the data of our hospital regarding epidemiology of the musculoskeletal system tumors and those from the other regions around the world. Level IV, Therapeutic study.


Electron capture induced dissociation of dipeptide dications: where does the charge go?
Charge partitioning after electron capture induced dissociation of dipeptide dications is determined by proton mobility in the evanescent ion-molecule complex as the remaining proton has enough time to choose the fragment with the highest proton affinity.


Molecular basis of dengue virus serotype 2 morphological switch from 29°C to 37°C.
The ability of DENV2 to display different morphologies (hence different antigenic properties) complicates vaccine and therapeutics development. Previous studies showed most strains of laboratory adapted DENV2 particles changed from smooth to "bumpy" surfaced morphology when the temperature is switched from 29°C at 37°C. Here we identified five envelope (E) protein residues different between two alternative passage history DENV2 NGC strains exhibiting smooth or bumpy surface morphologies. Several mutations performed on the smooth DENV2 infectious clone destabilized the surface, as observed by cryoEM. Molecular dynamics simulations demonstrated how chemically subtle substitution at various positions destabilized dimeric interactions between E proteins. In contrast, three out of four DENV2 clinical isolates showed a smooth surface morphology at 37°C, and only at high fever temperature (40°C) did they become "bumpy". These results imply vaccines should contain particles representing both morphologies. For prophylactic and therapeutic treatments, this study also informs on which types of antibodies should be used at different stages of an infection, i.e., those that bind to monomeric E proteins on the bumpy surface or across multiple E proteins on the smooth surfaced virus.


[An analysis of familial prostate cancer].
Clinical and pathological features of familial prostate cancer patients were examined. We compared 7 pairs (14 patients) of familial prostate cancer patients (Group F) with those of 1741 prostate cancer patients diagnosed in around Gunma Prefecture between 1987 and 1993 (Group G). Relative survival rates were compared because the average age was different in two groups. In Group F and Group G, age at diagnosis ranged from 54 to 86 and 47 to 97 years old, and the averages were 68.1 +/- 8.5 (S.D.) and 74.2 +/- 8.3, respectively. The average age tended to be younger in Group F. The ratio of early stage cancer tended to be higher and that of poorly differentiated cancer tended to be lower in Group F. The 3- and 5-year relative survival rate was 82.4% and 57.6% in Group F, and 84.3% and 73.9% in Group G, respectively. The 5-year survival rate tended to be lower in Group F, but almost no difference was observed between two groups on the whole. In Group F, of 6 patients whose causes of death were clear, 4 (66.7%) died of prostate cancer, while in Group G, of 398 patients whose causes of death were elaar, 224 (56.3%) died of prostate cancer. Family history of other cancer sites was positive in 3 of 6 families in Group F. Familial prostate cancer patients tended to be young, in early stage and low ratio of poorly differentiated cancer.


Phytoplankton morpho-functional trait dataset from French water-bodies.
In lake ecosystems, phytoplankton communities can be studied by adopting taxonomic-based approaches. However, these approaches suffer from identification issues and are sometimes of limited ecosystem ecological value. The recent development of functional approaches may allow an evaluation of other aspects of ecosystem quality, functions and interactions with abiotic parameters or other communities. Here, our aim was to create a phytoplankton trait database at the French scale. This database will be relevant for the analysis of phytoplankton communities that will lead to a better understanding of phytoplankton functional ecology in lakes of France and other European countries possessing similar biological communities. We used a French national database of phytoplankton occurrences sampled from 384 lakes over the entire French metropolitan territory. A final list of 636 taxa was used to compile 53 morpho-functional traits associated with taxonomic information. The traits encompassed variables such as shape, biovolume, motility, toxin production and Reynolds groups. With this new database, we aim to provide data for morpho-functional analyses of phytoplankton assemblages from local to European scale.


Improved delivery of radiolabeled anti-B1 monoclonal antibody to Raji lymphoma xenografts by predosing with unlabeled anti-B1 monoclonal antibody.
A human B-cell lymphoma xenograft model was used to test whether the administration of unlabeled MoAb prior to injection of radiolabeled monoclonal antibody (MoAb) improves delivery of the radiolabeled MoAb to tumor prior to testing in clinical radioimmunotherapy trials. The anti-B1/CD20 pan-B-cell MoAb reactive with human B-cell lymphomas and leukemias but not reactive with mouse B-cells was used in this study. Athymic nude mice bearing human Raji Burkitt lymphoma xenografts were given injections of 2.5 muCi (0.3 microgram) 131I-labeled anti-B1 with or without a 2-h prior single injection of 100 micrograms of unlabeled anti-B1 antibody. Four days later the animals given injections of 131I-labeled anti-B1 and the unlabeled anti-B1 predose had a tumor uptake of 12.72 +/- 1.17% (SEM) of injected dose/g which was 44% greater than the animals receiving the 131I-labeled anti-B1 alone (P = 0.014). The uptake in most normal tissues was unchanged, although the blood level of 131I-labeled anti-B1 appeared to be greater following unlabeled anti-B1 predosing (P = 0.067). Predosing with isotype matched irrelevant MoAb did not result in a greater tumor uptake or blood concentration of 131I-labeled anti-B1 compared to the administration of 131I-labeled anti-B1 alone. In studies using 111In-labeled anti-B1, the effect of unlabeled antibody predosing was more pronounced. For animals given injections of 4.5 muCi (0.4 microgram) 111In-labeled anti-B1 and the unlabeled anti-B1 predose, the uptake in tumor was 12.37 +/- 2.07% of injected dose/g which was 162% greater than the animals receiving the 111In-labeled anti-B1 alone (P = 0.009). Predosing decreased 111In-labeled anti-B1 uptake in spleen, while the blood level was significantly greater. Predosing was more effective than simultaneous injection in improving tumor delivery. When tumor-bearing mice were either simultaneously given injections of 36 micrograms of unlabeled anti-B1 and 4 micrograms 111In-labeled anti-B1 or were given preinjections of 36 micrograms unlabeled anti-B1 3 h prior to injection of 4 micrograms 111In-labeled anti-B1, tumor uptake 3 days later was 1.3-fold higher in the animals which received the preinjection of unlabeled antibody (P = 0.011). As the quantity of unlabeled anti-B1 was increased (36, 96, 996 micrograms) in the predose, significantly greater uptake in tumor was observed, although this uptake appeared to plateau at the highest predoses.(ABSTRACT TRUNCATED AT 400 WORDS)


Determination of DNA methylation using electrochemiluminescence with surface accumulable coreactant.
Cytosine methylation in DNA was determined by an enzyme linked immunosorbent assay (ELISA) with electrochemiluminescence (ECL) detection and employed for the DNA methylation assay of a long and real genomic sample for the first time. The developed method employed an antimethyl cytosine antibody labeled with acetylcholinesterase, which was added to recognize single methylated cytosine in a DNA oligomer. The acetylcholinesterase converted acetylthiocholine (substrate) to thiocholine (product), which was accumulated on a gold electrode surface via gold-thiol binding. This surface accumulated preconcentration made it possible to observe bright and distinctive ECL by applying a potential to the gold electrode in the presence of a tris(2,2-bipyridyl)ruthenium complex luminophore when the analyte DNA contained a methylation region. Methyl-cytosine was measured quantitatively in the 1-100 pmol range, which exhibits sufficiently high sensitivity to achieve real DNA measurements without amplification by a polymerase chain reaction (PCR). The proposed ECL method also exhibited high selectivity for methyl-cytosine against nonmethylated cytosine, guanine, thymine, and adenine nucleotides. Finally, original and methylated DNA samples were clearly distinguished with our method using a real DNA bacteriophage sample (48,502 base pairs).


[Polymorphism of KPI-A genes from plants of the subgenus Potatoe (sect. Petota, Estolonifera and Lycopersicum) and subgenus Solanum].
Kunitz-type proteinase inhibitor proteins of group A (KPI-A) are involved in the protection of potato plants from pathogens and pests. Although sequences of large number of the KPI-A genes from different species of cultivated potato (Solanum tuberosum subsp. tuberosum) and a few genes from tomato (Solanum lycopersicum) are known to date, information about the allelic diversity of these genes in other species of the genus Solanum is lacking. In our work, the consensus sequences of the KPI-A genes were established in two species of subgenus Potatoe sect. Petota (Solanum tuberosum subsp. andigenum--5 genes and Solanum stoloniferum--2 genes) and in the subgenus Solanum (Solanum nigrum--5 genes) by amplification, cloning, sequencing and subsequent analysis. The determined sequences of KPI-A genes were 97-100% identical to known sequences of the cultivated potato of sect. Petota (cultivated potato Solanum tuberosum subsp. tuberosum) and sect. Etuberosum (S. palustre). The interspecific variability of these genes did not exceed the intraspecific variability for all studied species except Solanum lycopersicum. The distribution of highly variable and conserved sequences in the mature protein-encoding regions was uniform for all investigated KPI-A genes. However, our attempts to amplify the homologous genes using the same primers and the genomes of Solanum dulcamarum, Solanum lycopersicum and Mandragora officinarum resulted in no product formation. Phylogenetic analysis of KPI-A diversity showed that the sequences of the S. lycopersicum form independent cluster, whereas KPI-A of S. nigrum and species of sect. Etuberosum and sect. Petota are closely related and do not form species-specific subclasters. Although Solanum nigrum is resistant to all known races of economically one of the most important diseases of solanaceous plants oomycete Phytophthora infestans aminoacid sequences encoding by KPI-A genes from its genome have nearly or absolutely no differences to the same from genomes of cultivated potatoes involved by P. infestans.


Serum dioxin, insulin, fasting glucose, and sex hormone-binding globulin in veterans of Operation Ranch Hand.
We studied insulin, fasting glucose, and sex hormone-binding globulin (SHBG) in Air Force veterans exposed to Agent Orange and its contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin), during the Vietnam War. The index subjects were veterans of Operation Ranch Hand, the unit responsible for aerial herbicide spraying in Vietnam from 1962-1971. Other Air Force veterans who served in Southeast Asia during the same period but were not involved with spraying herbicides served as comparisons. We assigned each Ranch Hand veteran based on his dioxin level to one of three exposure categories, named background, low, and high. Among nondiabetic veterans, we found the mean of the logarithm of insulin significantly increased in the high dioxin category. Additionally, in nondiabetic veterans the relation between SHBG and insulin interacted significantly with dioxin category on the log scale within strata defined by age and percent body fat. Among young (age, < or = 53 yr), lean (percent body fat, < or = 25%) nondiabetic veterans in the high category, the slope relating the logarithm of SHBG and the logarithm of insulin was significantly decreased. These findings suggest a compensatory metabolic relationship between dioxin and insulin regulation.


List-decoding methods for inferring polynomials in finite dynamical gene network models.
The problem of reverse engineering the dynamics of gene expression profiles is of focal importance in systems biology. Due to noise and the inherent lack of sufficiently large datasets generated via high-throughput measurements, known reconstruction frameworks based on dynamical systems models fail to provide adequate settings for network analysis. This motivates the study of new approaches that produce stochastic lists of explanations for the observed network dynamics that can be efficiently inferred from small sample sets and in the presence of errors. We introduce a novel algebraic modeling framework, termed stochastic polynomial dynamical systems (SPDSs) that can capture the dynamics of regulatory networks based on microarray expression data. Here, we refer to dynamics of the network as the trajectories of gene expression profiles over time. The model assumes that the expression data is quantized in a manner that allows for imposing a finite field structure on the observations, and the existence of polynomial update functions for each gene in the network. The underlying reverse engineering algorithm is based on ideas borrowed from coding theory, and in particular, list-decoding methods for so called Reed-Muller codes. The list-decoding method was tested on synthetic data and on microarray expression measurements from the M(3D) database, corresponding to a subnetwork of the Escherichia coli SOS repair system, as well as on the complete transcription factor network, available at RegulonDB. The results show that SPDSs constructed via list-decoders significantly outperform other algebraic reverse engineering methods, and that they also provide good guidelines for estimating the influence of genes on the dynamics of the network. Software codes for list-decoding algorithms suitable for direct application to quantized expression data will be publicly available at the authors' web-pages.


Consequences of using escharotic agents as primary treatment for nonmelanoma skin cancer.
The use of escharotic or caustic pastes to treat skin cancer is based on the centuries-old observation that selected minerals and plant extracts may be used to destroy certain skin lesions. Zinc chloride and Sanguinaria canadensis (bloodroot) are 2 agents that are used as part of the Mohs chemosurgery fixed-tissue technique. The use of escharotics without surgery has been discredited by allopathic medicine but persists and is promoted among alternative practitioners. Patients may now purchase "herbal supplements" for the primary self-treatment of skin cancer, and physicians will see patients who elect this therapy for their skin cancers. We reviewed the history of escharotic use for skin disease and performed an Internet search for the availability and current use of escharotics. Our search located numerous agents for purchase via the Internet that are advertised as highly successful treatments for skin cancer. We report 4 cases from our practice in which escharotic agents were used by patients to treat basal cell carcinomas in lieu of the recommended conventional treatment. One patient had a complete clinical response, but had a residual tumor on follow-up biopsy. A second patient successfully eradicated all tumors, but severe scarring ensued. A third patient disagreed with us regarding his care and was lost to follow-up. One patient presented with a nasal basal cell carcinoma that "healed" for several years following treatment elsewhere with an escharotic agent but recurred deeply and required an extensive resection. The lesion has since metastasized. Escharotic agents are available as herbal supplements and are being used by patients for the treatment of skin cancer. The efficacy of these agents is unproven and their content is unregulated. Serious consequences may result from their use. Conventional medicine has an excellent track record in treating skin cancer. Physicians should recommend against the use of escharotic agents for skin cancer, and the Food and Drug Administration should be given the authority to regulate their production and distribution.


In vitro sunscreen transmittance measurement with concomitant evaluation of photostability: evolution of a method.
The recent paper by Miura et al. (Photochem. Photobiol. 84[6], 1569-1575) offers a re-examination of extant in vitro methods for dynamically measuring sunscreen photodegradation under continuous irradiation in situ. We commend the authors' efforts toward developing an improved system for accurate in vitro sunscreen assessment. This work describes an alternate derivative apparatus incorporating an improved detector which may prove an exceptionally valuable contribution toward that goal. Unfortunately their report suffers from insufficient detail in instrumentation description and lacks requisite calibration procedures. Their utilization of a solar simulator filtered for conventional in vivo sun protection factor (SPF) testing poses transmittance measurement limitations at short wavelengths that are not adequately addressed and is also deficient, relative to sunlight, in longer UVA wavelengths shown to contribute to sunscreen photoinstability. We concur that the in vitro sunscreen testing should utilize continuous or multiple irradiation doses and should ideally use the same 2 mg cm(-2) product application amount as does the human SPF test. We encourage their proposal that methodology, which simultaneously measures sunscreen spectral transmittance and photodegradation under continuous irradiation to an accumulated erythemic endpoint, as we previously described, be developed into a consensus test standard.


Continuous tooth eruption in Australian aboriginal skulls.
Increases in the distance from the cemento-enamel junction to the alveolar crest (CEJ-AC) have often been attributed to senile atrophy of the bone or to the effects of periodontitis, without reference to the condition of the alveolar crest. This study investigated the relationship between CEJ-AC distance, tooth wear, gender, site of the CEJ-AC measurements, and age in 161 pre-white-contact Australian aboriginal skulls. Individual teeth were included in the study when there was no evidence of dehiscence, periodontitis, or abscess cavity formation in the supporting alveolar bone. The CEJ-AC distances increased as the severity of attrition increased; in male skulls, CEJ-AC distances were greater than in female skulls for all categories of tooth wear. In general, CEJ-AC distances measured on the mesial aspects of teeth were greater than those recorded distally; lingual distances generally exceeded buccal recordings. The best explanation of these findings and similar reports in the anthropological literature is that continuous tooth eruption occurred without the concomitant coronal movement of alveolar bone. This conclusion has significant ramifications for anthropological, epidemiological, and clinical studies which use the CEJ and AC as reference points when measuring periodontal attachment loss (periodontitis).


Tissue microarray molecular profiling of early, node-negative adenocarcinoma of the rectum: a comprehensive analysis.
Early-stage adenocarcinoma of the rectum treated with curative intent has a favorable overall prognosis; however, 20%-30% of the patients recur, and the majority ultimately die of disease. Recurrence and tumor-related mortality may be attributable to molecular abnormalities in primary tumors accounting for their more aggressive biological behavior. This study evaluates such molecular phenotypes with regard to cell cycle regulation and proliferation and determines their significance for patient outcome. One hundred patients with primary T(2-3), N(0) adenocarcinoma of the rectum uniformly treated by surgery alone were studied. Core biopsies of pathological specimens were assembled on tissue microarrays, and expression of p53, mdm-2, p21, Bcl-2, p27, cyclin D1, and Ki-67 was analyzed by immunohistochemistry. Molecular profiles were correlated with disease-free (DFS) and disease-specific survival (DSS). Despite previously described prognostic relevance of some of the investigated molecules in analyses where different stages of colorectal cancer were included, none of the cell cycle-regulatory or proliferation-related markers was associated with recurrence or survival. However, patients with tumors demonstrating down-regulation of p27, a cyclin-dependent kinase inhibitor and tumor suppressor gene associated with development of metastases, showed a trend toward reduced DFS and DSS (P = 0.06 and P = 0.07, respectively). In this homogeneous group of patients with early-stage, node-negative adenocarcinoma of the rectum uniformly treated by surgery alone, the investigated cell cycle-regulatory and proliferation-associated proteins appear to have no prognostic significance. However, down-regulation of p27 appears to be associated with a trend toward reduced DFS and DSS, which suggests further investigation of other p27-related pathways potentially relevant for metastatic disease.


Design and optimization of a total vaporization technique coupled to solid-phase microextraction.
Solid-phase microextraction (SPME) is a popular sampling technique in which chemical compounds are collected with a sorbent-coated fiber and then desorbed into an analytical instrument such as a liquid or gas chromatograph. Typically, this technique is used to sample the headspace above a solid or liquid sample (headspace SPME), or to directly sample a liquid (immersion SPME). However, this work demonstrates an alternative approach where the sample is totally vaporized (total vaporization SPME or TV-SPME) so that analytes partition directly between the vapor phase and the SPME fiber. The implementation of this technique is demonstrated with polydimethylsiloxane-divinylbenzene (PDMS-DVB) and polyacrylate (PA) coated SPME fibers for the collection of nicotine and its metabolite cotinine in chloroform extracts. The most important method parameters were optimized using a central composite design, and this resulted in an optimal extraction temperature (96 °C), extraction time (60 min), and sample volume (120 μL). In this application, large sample volumes up to 210 μL were analyzed using a volatile solvent such as chloroform at elevated temperatures. The sensitivity of TV-SPME is nearly twice that of liquid injection for cotinine and nearly 6 times higher for nicotine. In addition, increased sampling selectivity of TV-SPME permits detection of both nicotine and cotinine in hair as biomarkers of tobacco use where in the past the detection of cotinine has not been achieved by conventional SPME.


[Simultaneous determination of 121 veterinary drugs in pork by QuEChERS and ultra performance liquid chromatography-tandem mass spectrometry].
A method has been developed for the simultaneous determination of the 10 kinds of 121 veterinary drugs (including chloramphenicols, β-agonists, sulfonamides, 4-quinolones, nitroimidazoles, macrolides, avermections, polyether antibiotics, tetracycline antibiotics, cephalosporins, etc.) in pork using QuEChERS method coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The drugs were extracted with Na2EDTA-McIlvaine buffer solution (pH = 4) and acetonitrile. Anhydrous sodium sulfate was used for the salting-out process. The solutions divided into two groups were cleaned-up by different mixed sorbents. The drugs were analyzed by UPLC-MS/MS in multiple reaction monitoring (MRM) mode via electrospray ionization. Among the 121 drugs, 5 drugs were quantified by internal standard method, and the other 116 drugs were quantified by external standard method. The calibration curves of the 121 drugs were linear in the range of 0.02-40 μg/L with the correlation coefficients more than 0.99. The limits of quantification (LOQs, S/N = 10) were 0.05-10 μg/kg. The average recoveries of 8 drugs at LOQ level in pork ranged from 41.7% to 59.6% with the relative standard deviations (RSDs) of 1.7% - 13.5%. The average recoveries of 10 drugs at LOQ level in pork ranged from 122.6% to 163.2% with the RSDs of 0.6% -14.8%. The average recoveries of 103 drugs at LOQ level in pork ranged from 60.3% to 118.3% with the RSDs of 0.4% - 16.7%. The proposed method is rapid, simple, sensitive, reliable and cost-effective.


Effects of brussels sprouts and their phytochemical components on oxidative stress-induced neuronal damages in PC12 cells and ICR mice.
In this study, the protective effects of Brussels sprouts extract and its major constituents against oxidative stress-induced damages were investigated in rat pheochromocytoma cells and Institute of Cancer Research mice. The major constituents of Brussels sprouts (3,4',5,7-tetrahydroxyflavone (kaempferol), indole-3-carbinol, and phenethyl isothiocyanate) were selectively tested. Of these, the flavonoid compound, kaempferol exhibited the highest potency in radical scavenging activity (1,1-diphenyl-2-picryl hydrazyl assay and oxygen radical absorbance capacity assay) and was most protective against oxidative stress in neuronal cell assays (measurement of intracellular oxidative stress levels and cell viability). In mice, after 4 weeks of kaempferol administration, significant protection against amyloid beta (Aβ) peptide-induced neurotoxicity was also observed, as assessed through the passive avoidance test. Taken together, the results suggest that Brussels sprouts could be protective against Aβ-induced neurotoxicity, possibly due to the antioxidative capacity of its major constituent, kaempferol.


Subcutaneous phaeohyphomycosis of foot in an immunocompetent host.
We report a case of subcutaneous phaeohyphomycosis of foot, which is a mycotic disease that produces brown pigmented hyphae, pseudohyphae and yeast form in combination. The patient was immunocompetent and had injury 23 years before, and developed a non healing foot ulcer which was clinically suspected as tuberculous or carcinomatous etiology. Local wide excision was done and sent in formalin for histopathological examination. Microscopically pigmented septate, hyphae and yeast forms were identified on hematoxylin and eosin stained sections which were confirmed by special stains such as periodic acid-Schiff and Gomori's methenamine silver stains.


A survey of findings on the economic and regulatory determinants of the demand for alcoholic beverages.
This paper summarizes the findings of two studies on the demand for alcoholic beverages. The first study surveyed the results of over twenty demand estimation studies conducted between 1945 and 1977, covering many countries. The second study estimated the demand for distilled spirits and beer across states and the District of Columbia for the period 1974-1978. The main finding is that among all the regulatory variables available to reduce mean consumption, price has the strongest impact.


Long-term dechlorination of cis-DCE to ethene with co-immobilized Dehalococcoides mccartyi BAV1 and Clostridium butyricum in silica gel system.
Chloroethenes are common groundwater pollutants, and have been classified as toxic and carcinogenic to humans. The metabolites of chloroethenes, cis-dichloroethene (cis-DCE) and vinyl chloride (VC) commonly accumulate in groundwater due to their recalcitrant reductive dechlorination under anaerobic conditions. Dehalococcoides mccartyi (Dhc) is the key anaerobic bacteria for complete dechlorination of chloroethene, and Clostridium butyricum (C. butyricum) can provide hydrogen for supporting the growth of Dhc. In this study, we co-immobilized Dhc strain BAV1 and C. butyricum in a silica gel to determine the ability of the complete dechlorination of cis-DCE. Our results showed that our immobilized system could protect BAV1 from a high concentration (8 mM) of cis-DCE to carry out complete dechlorination. After the long-term use of our immobilized system, the activity of complete dechlorination was maintained for more than 180 consecutive days. Furthermore, we applied the immobilized system to remediate contaminated groundwater and uncovered the complete dechlorination of cis-DCE into ethene, a non-toxic product, within 28 days. Therefore, this novel co-immobilized system could serve a solution for bioremediation at chloroethene-contaminated sites.


Polypeptide from Chlamys farreri restores endoplasmic reticulum (ER) redox homeostasis, suppresses ER stress, and inhibits ER stress-induced apoptosis in ultraviolet B-irradiated HaCaT cells.
To investigate the effects of polypeptide from Chlamys farreri (PCF) on ultraviolet B (UVB)-induced apoptosis in human keratinocyte HaCaT cells. In HaCaT cells at 4 h or 18 h after UVB irradiation, the cell viability was measured by MTT assay. Cellular apoptosis was detected with annexin V-FITC/PI staining by flow cytometry. The expression levels of PDI, Ero-1α, GRP78, and CHOP were assessed by Western blot analysis. Mitochondrial membrane potential (MMP) was measured by fluorescent probe JC-1. Caspase activities were detected with fluorogenic substrates. PCF alleviated cell viability loss and inhibited apoptosis in HaCaT cells after UVB irradiation. Moreover, PCF increased the expression levels of PDI and Ero-1α, which were related with the ER redox homeostasis. Furthermore, PCF treatment inhibited the expression of GRP78 at 4 h after UVB irradiation, and suppressed CHOP expression at 18 h post-irradiation, indicating that PCF could inhibit UVB-evoked ER stress in the early stage post-irradiation, and suppress the ER stress-induced apoptosis in the late stage. In addition, PCF alleviated UVB-induced MMP loss, and inhibited the activation of caspase-9/-3, in HaCaT cells after UVB irradiation. On the other hand, MMP loss and caspase-9/-3 activation could be partly blocked by the ER stress inhibitor 4-PBA. PCF inhibits UVB-induced apoptosis through restoring ER redox homeostasis, suppressing ER stress, and inhibiting ER stress-induced mitochondrial apoptosis in HaCaT cells. These findings provide evidence for the mechanism underlying UVB-induced skin damages, and support the promising role of PCF in treatment of the diseases.


Late life function and disability instrument: I. Development and evaluation of the disability component.
Efforts to evaluate the effectiveness of clinical and community-based interventions designed to impact late-life disability have been hindered significantly by limitations in current instrumentation. More conceptually sound and responsive measures of disability are needed. Applying Nagi's disablement model, we wrote questionnaire items that assessed disability in terms of frequency and limitation in performance of 25 life tasks. We evaluated their validity and test-retest reliability with 150 ethnically and racially diverse adults aged 60 and older who had a range of functional limitations, using factor analysis and Rasch analytic techniques to examine and refine the instrument. Our analyses resulted in a 16-item disability component with two dimensions, one focused on frequency of performance and the other addressing limitation in performance of life tasks, with two disability domains within each dimension. The frequency dimension consisted of a personal and a social role domain, and the limitation dimension consisted of an instrumental and a management role domain. Expected differences in summary scores of known-functional limitation groups support the validity of this instrument. Test-retest intraclass correlations of the reproducibility of each overall dimension summary score were moderate to high (intraclass correlation coefficients .68-.82). The Late-Life Function and Disability Instrument has potential to assess meaningful concepts of disability across a wide variety of life tasks with relatively few items.


Prevalence and significance of prominent mucin pools in the esophagus post neoadjuvant chemoradiotherapy for Barrett's-associated adenocarcinoma.
Occasionally, patients with Barrett's-associated adenocarcinoma who received preoperative chemoradiotherapy (chemrad) show prominent mucin pools in their resection specimen, but the prognostic significance of this finding has never been investigated. The purpose of this study was to evaluate the clinical and pathologic features, and prognostic significance, of prominent mucin pools in 21 patients identified from a cohort of 192 consecutive cancer patients (prevalence rate, 10.9%) who had an esophagectomy (post-chemrad) for adenocarcinoma. The clinical and pathologic features and follow-up data were evaluated in 21 patients with adenocarcinoma of the esophagus and with prominent mucin pools (male-to-female ratio, 21:0; mean age, 61 years) and compared with a control group of 19 consecutive chemrad-treated and stage-matched esophageal adenocarcinoma patients who had either minimal microscopic (9 cases) or no (10 cases), residual disease present in their resection specimen (male-to-female ratio, 18:1; mean age, 62 years). Of the 21 study patients, 7 (33%) had acellular mucin pools with no residual tumor, 7 (33%) showed rare isolated tumor cells within mucin pools, and 7 (33%) had acellular mucin pools with microscopic foci of residual adenocarcinoma in tissue adjacent to, but not within, mucin pools. In total, 2 cases (9%) showed mucin pools limited to the submucosa, 5 (24%) showed involvement of the muscularis propria, and 14 (67%) showed mucin pools within the muscularis propria and adventitia. Four cases (19%) showed involvement of the radial resection margin with acellular mucin pools. Thirteen study patients (62%) contained acellular mucin pools within regional lymph nodes. A significantly higher proportion of patients with prominent mucin pools had mucinous adenocarcinoma prior to neoadjuvant therapy, compared with control patients without mucin pools (12 of 21 (57%) vs. 1 of 19 (5%), P < 0.001). Upon follow-up (mean, 27 months; range, 5-84 months), none of the study patients with prominent mucin pools died of disease (0%), in comparison to 5 of 19 (26%) control patients (mean follow-up, 27 months; range, 3-64 months) (P = 0.02). None of the patients who had acellular mucin pools involving the radial margin developed recurrence or metastasis or died of disease. Prominent mucin pools in resection specimens from patients with Barrett's esophagus-associated adenocarcinoma who received preoperative chemrad are associated with mucinous tumors and are not associated with poor survival, even when acellular mucin pools involve the radial margin, and, thus, should not be interpreted as evidence of residual viable adenocarcinoma.


Risk, human error, and system resilience: fundamental ideas.
I review and critique basic ideas of both traditional error/risk analysis and the newer and contrasting paradigm of resilience engineering. Analysis of human error has matured and been applied over the past 50 years by human factors engineers, whereas the resilience engineering paradigm is relatively new. Fundamental ideas and examples of human factors applications of each approach are presented and contrasted. Probabilistic risk analysis provides mathematical rigor in generalizing on past error events to identify system vulnerabilities, but prediction is problematical because (a) error definition is arbitrary, and thus it is difficult to infer valid probabilities of human error to input to quantitative models, and (b) future accident conditions are likely to be quite different from those of past accidents. The new resilience engineering paradigm, in contrast, is oriented toward organizational process and is concerned with anticipating, mitigating, and preparing for graceful recovery from future events. Resilience engineering complements traditional error analysis but has yet to provide useful quantification and operational methods. A best safety strategy is to use both approaches.


The Influence of Proton Pump Inhibitors on the Fecal Microbiome of Infants with Gastroesophageal Reflux-A Prospective Longitudinal Interventional Study.
Proton pump inhibitors (PPIs) are the standard therapy for gastroesophageal reflux disease. In adults, PPI treatment is associated with Clostridium difficile infections (CDI). In contrast to adults the microbiome of infants develops from sterility at birth toward an adult-like profile in the first years of life. The effect of PPIs on this developing microbiome has never been studied. The aim of the present study was to determine the effect of oral PPIs on the fecal microbiome in infants with gastroesophageal reflux disease (GERD). In this prospective longitudinal study 12 infants with proven GERD received oral PPIs for a mean period of 18 weeks (range 8-44). Stool samples were collected before ("before PPI") and 4 weeks after initiation of PPI therapy ("on PPI"). A third sample was obtained 4 weeks after PPI discontinuation ("after PPI"). The fecal microbiome was determined by NGS based 16S rDNA sequencing. This trial was registered with clinicaltrials.gov (NCT02359604). In a comparison of "before PPI" and "on PPI" neither α- nor β-diversity changed significantly. On the genus level, however, the relative abundances showed a decrease of Lactobacillus and Stenotrophomonas and an increase of Haemophilus. After PPI therapy there was a significant increase of α- and β-diversity. Additionally, the relative abundances of the phyla Firmicutes, Bacteroidetes, and Proteobacteria were significantly changed and correlated to patients' age and the introduction of solid foods. PPI treatment has only minor effects on the fecal microbiome. After discontinuation of PPI treatment the fecal microbiome correlated to patients' age and nutrition.


Comparison of tracer techniques for monitoring sewer losses.
Two different experimental techniques employing conservative tracers to determine exfiltration losses in a sewer system are reported. Both techniques compare the in-sewer responses measured along designated indicator and reference sections to assess the level of exfiltration although with different degrees of reliability. The experimental and interpretation errors associated with a pulse dosing technique, in which Rhodamine WT is used as the single tracer, are described. The estimation of exfiltration requires the comparison of indicator and reference peaks which need to be well resolved and reproducible in order to provide realistic results. A more consistent technique involves the continuous dosing of two different tracers with the objective of producing simultaneous downstream peak tracer concentrations. Any errors due to flow variations are eliminated and the longer measurement period allows both peak and background concentrations to be more accurately determined. Rhodamine WT, Li and Br were used in different combinations over the approximately 20 min duration of each continuous dosing experiment and were found to demonstrate conservative behaviours with measured recoveries of 97.6-100.6%, 97.3-101.4% and 108.4-109.2%, respectively. The analytical problems associated with the determination of the different tracers in sewage are described and are shown to be responsible for the reduced accuracy of the results associated with the use of Br. The preferred combination of tracers is Rhodamine WT as the indicator and Li as the reference and these are shown to be capable of estimating sewer losses to an accuracy of +/-1.0% using the continuous dosing approach.


Different types of nsP3-containing protein complexes in Sindbis virus-infected cells.
Alphaviruses represent a serious public health threat and cause a wide variety of diseases, ranging from severe encephalitis, which can result in death or neurological sequelae, to mild infection, characterized by fever, skin rashes, and arthritis. In the infected cells, alphaviruses express only four nonstructural proteins, which function in the synthesis of virus-specific RNAs and in modification of the intracellular environment. The results of our study suggest that Sindbis virus (SINV) infection in BHK-21 cells leads to the formation of at least two types of nsP3-containing complexes, one of which was found in association with the plasma membrane and endosome-like vesicles, while the second was coisolated with cell nuclei. The latter complexes could be solubilized only with the cytoskeleton-destabilizing detergent. Besides viral nsPs, in the mammalian cells, both complexes contained G3BP1 and G3BP2 (which were found in different ratios), YBX1, and HSC70. Rasputin, an insect cell-specific homolog of G3BP1, was found in the nsP3-containing complexes isolated from mosquito cells, which was suggestive of a high conservation of the complexes in the cells of both vertebrate and invertebrate origin. The endosome- and plasma membrane-associated complexes contained a high concentration of double-stranded RNAs (dsRNAs), which is indicative of their function in viral-RNA synthesis. The dsRNA synthesis is likely to efficiently proceed on the plasma membrane, and at least some of the protein-RNA complexes would then be transported into the cytosol in association with the endosome-like vesicular organelles. These findings provide new insight into the mechanism of SINV replication and virus-host cell interactions.


[The role of inhaled steroids in the treatment of COPD].
The aim of COPD treatment is to reduce symptoms and prevent exacerbations. The pharmacological treatment is based on bronchodilator treatment which reduces symptoms and prevents exacerbations. If patients have exacerbations despite bronchodilator treatment it is important to consider adding inhaled corticosteroids (ICS). Concomitant asthma is always an indication for ICS treatment. Studies have shown that measurement of blood eosinophil count predicts the effect of adding ICS to bronchodilators. Higher b-eos (≥ 0,3 × 109/l) strengthens the indication for ICS. A large number of COPD patients in Sweden are treated with both bronchodilators and ICS without a history of exacerbations. It is important to assess that the indication is correct at follow up of these patients.


Distribution of ventilation/perfusion ratios in pulmonary embolism: an adjunct to the interpretation of ventilation/perfusion lung scans.
Diagnosis of pulmonary embolism (PE) by visual interpretation of ventilation/perfusion (V/Q) scans is limited by the high percentages of patients classified in the intermediate- and low-probability categories. This study proposes a quantitative analysis of the distribution of V/Q ratios to better identify patients with PE. We studied 99 consecutive patients who underwent dual-isotope (81m)Kr/(99m)Tc-macroaggregate V/Q scanning and arterial blood gas analysis within 48 h. The 8-view V/Q scans were visually analyzed by 2 observers according to the revised criteria of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) (normal scan or low, intermediate, or high probability of PE). Quantitative analysis of the posterior-view distribution histogram of V/Q ratios was performed using dedicated software. Briefly, regions of interest were drawn around the lungs on the matched V/Q images, smooth filtering was applied, normalized regional V/Q ratios were calculated within each pixel, and a distribution histogram was built. Patients with normal scans (n = 16) had a predominance of V/Q ratios (63.3% +/- 13.0%) between 0.8 and 1.2. They had only 9.8% +/- 5.8% of ratios > 1.2, and the remaining 26.9% +/- 7.5% of ratios were <0.8. By contrast, patients with PE (n = 34) were characterized by a significant increase (15.5 +/- 10.0%, P = 0.04) in high V/Q ratios (>1.2) and a significant increase (34.5% +/- 8.2%, P = 0.003) in low V/Q ratios (<0.8). Interestingly, a similar pattern was found in patients with a high PIOPED probability of PE, 21.3% +/- 11.0% and 37.5% +/- 9.2%, respectively. Within the nondiagnostic group (intermediate- + low-probability scans, n = 58), 17 patients were finally diagnosed with PE. Analysis of the distribution histogram in this group allowed the identification of 5 patients with PE (specificity, 78%). A quantitative approach to lung scan interpretation, based on the distribution histogram of V/Q ratios, may be helpful for categorizing patients with suspected PE.


Manganese co-localizes with calcium and phosphorus in Chlamydomonas acidocalcisomes and is mobilized in manganese-deficient conditions.
Exposing cells to excess metal concentrations well beyond the cellular quota is a powerful tool for understanding the molecular mechanisms of metal homeostasis. Such improved understanding may enable bioengineering of organisms with improved nutrition and bioremediation capacity. We report here that Chlamydomonas reinhardtii can accumulate manganese (Mn) in proportion to extracellular supply, up to 30-fold greater than its typical quota and with remarkable tolerance. As visualized by X-ray fluorescence microscopy and nanoscale secondary ion MS (nanoSIMS), Mn largely co-localizes with phosphorus (P) and calcium (Ca), consistent with the Mn-accumulating site being an acidic vacuole, known as the acidocalcisome. Vacuolar Mn stores are accessible reserves that can be mobilized in Mn-deficient conditions to support algal growth. We noted that Mn accumulation depends on cellular polyphosphate (polyP) content, indicated by 1) a consistent failure of C. reinhardtii vtc1 mutant strains, which are deficient in polyphosphate synthesis, to accumulate Mn and 2) a drastic reduction of the Mn storage capacity in P-deficient cells. Rather surprisingly, X-ray absorption spectroscopy, EPR, and electron nuclear double resonance revealed that only little Mn2+ is stably complexed with polyP, indicating that polyP is not the final Mn ligand. We propose that polyPs are a critical component of Mn accumulation in Chlamydomonas by driving Mn relocation from the cytosol to acidocalcisomes. Within these structures, polyP may, in turn, escort vacuolar Mn to a number of storage ligands, including phosphate and phytate, and other, yet unidentified, compounds.


Imidazole, a New Tunable Reagent for Producing Nanocellulose, Part I: Xylan-Coated CNCs and CNFs.
Imidazole is reported to be an effective reactant for the production of nanocellulose from hardwood pulp. The morphologies and surface properties of the nanocellulose can be simply tailored according to the water content in the imidazole system: with pure imidazole, cellulose nanofibrils (CNFs) in a yield of 10 wt % can be produced. With 25 wt % of water in imidazole, cellulose nanocrystals (CNCs) are obtained in 20 wt % yield. Both nanocelluloses exhibit crystallinity indices in the order of 70%. Interestingly, they retain the original xylan from the pulp with ca. 9⁻10 wt % of residual xylan content.


Increasing Incidence and Unique Clinical Characteristics of Spinning-Induced Rhabdomyolysis.
To compare outcomes of spinning-induced rhabdomyolysis to those with exertional rhabdomyolysis from other physical activities. Retrospective cohort study. Academic medical center, single-center. A retrospective chart review was conducted on patients evaluated from December 2010 through November 2014. Patients were selected by ICD-9 code for rhabdomyolysis. Patients were included if the reason for admission was rhabdomyolysis caused by exertion. Cases of rhabdomyolysis caused by trauma or drugs were excluded. Muscle group involvement, admission, and peak creatine kinase levels, time from activity to hospitalization, length of hospital stay, and incidence of complications. Twenty-nine cases were reviewed with 14 admissions secondary to spinning. Median admission creatine kinase (73 000 IU/L vs 29 000 IU/L, P = 0.02) and peak creatine kinase levels were significantly higher in the spinning group (81 000 IU/L vs 31 000 IU/L, P = 0.007). Hospital admissions for spinning-induced rhabdomyolysis increased over time. Health care providers should be aware of the potential dangers of spinning-related rhabdomyolysis especially in otherwise healthy young people.


Integration of artificial neural network and physiologically based biopharmaceutic models in the development of sustained-release formulations.
Model-informed drug development is an important area recognized by regulatory authorities and is gaining increasing interest from the generic drug industry. Physiologically based biopharmaceutics modeling (PBBM) is a valuable tool to support drug development and bioequivalence assessments. This study aimed to utilize an artificial neural network (ANN) with a multilayer perceptron (MLP) model to develop a sustained-release matrix tablet of metformin HCl 500 mg, and to test the likelihood of the prototype formulation being bioequivalent to Glucophage® XR, using PBBM modeling and virtual bioequivalence (vBE). The ANN with MLP model was used to simultaneously optimize 735 formulations to determine the optimal formulation for Glucophage® XR release. The optimized formulation was evaluated and compared to Glucophage® XR using PBBM modeling and vBE. The optimized formulation consisted of 228 mg of hydroxypropyl methylcellulose (HPMC) and 151 mg of PVP, and exhibited an observed release rate of 42% at 1 h, 47% at 2 h, 55% at 4 h, and 58% at 8 h. The PBBM modeling was effective in assessing the bioequivalence of two formulations of metformin, and the vBE evaluation demonstrated the utility and relevance of translational modeling for bioequivalence assessments. The study demonstrated the effectiveness of using PBBM modeling and model-informed drug development methodologies, such as ANN and MLP, to optimize drug formulations and evaluate bioequivalence. These tools can be utilized by the generic drug industry to support drug development and biopharmaceutics assessments.


IL-1 and TNF-alpha are important factors in the pathogenesis of murine recurrent herpetic stromal keratitis.
To better understand the role of interleukin (IL)-1 and tumor necrosis factor (NF)alpha in recurrent herpetic stromal keratitis (HSK), the cytokine content and the effects of anti-cytokine antibodies on mouse corneas with the disease were examined. Competitive reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent analyses of IL-1alpha and TNF-alpha content were performed on corneas removed 3, 5, 7, 10, 14, and 21 days after latently infected NIH mice were irradiated with UV-B light to reactivate herpes simplex virus (HSV). In separate experiments, mice were injected with anti-IL-1 or anti-TNF-a antibodies 1 day before and 7 days after reactivation. UV-B irradiation stimulated an increase in corneal IL-la mRNA in reactivated (virus shedding) mice. This increase persisted longer and was higher than in UV-B irradiated uninfected control animals. IL-1alpha and TNF-alpha protein in corneas of reactivated mice was significantly elevated on days 3 to 10 compared with day 0 levels, and exceeded levels in control corneas on the same days. Anti-IL-1 and anti-TNF-alpha antibody administration both resulted in significantly decreased virus-induced corneal opacity between 7 and 21 days after UV-B exposure. IL-1alpha and TNF-alpha are upregulated in corneas in mice experiencing recurrent HSK. Abrogation of virus-induced corneal disease by anti-cytokine antibodies suggests that these cytokines play important roles in the pathogenesis of recurrent disease. Therefore, neutralization of specific proinflammatory cytokines may have potential therapeutic value.


Recovery times from subarachnoid blocks using bupivacaine hydrochloride and tetracaine hydrochloride with and without epinephrine.
This retrospective study examined the length of time patients spent in the postanesthesia care unit (PACU) recovering from a subarachnoid block with either bupivacaine hydrochloride or tetracaine hydrochloride with and without epinephrine after total knee replacement surgery or total hip replacement surgery. One hundred subjects' charts were reviewed with 50 subjects receiving a subarachnoid block with bupivacaine (25 had epinephrine added to the bupivacaine) and 50 subjects receiving a subarachnoid block with tetracaine (25 had epinephrine added to the tetracaine). There were no statistical differences among the groups with respect to age, height, weight, dose of local anesthetic, and length of surgical procedure. Patient who received tetracaine stayed longer in the PACU (64.44 minutes) and took longer to bend their knees (73.17 minutes), flex their hips (99.65 minutes), and have return of sensation (68.88 minutes), compared to those who had received bupivacaine (P < .05). When epinephrine was added to the local anesthetic, it prolonged the time until the return of knee flexion, hip flexion, and sensation by 66.82, 87.65, and 76.77 minutes respectively (P < .05).


Hyperemia-associated costs of medication changes in glaucoma patients treated initially with prostaglandin analogs.
To develop a model to estimate and compare the cost of changing therapy due to hyperemia in glaucoma patients treated initially either with latanoprost, bimatoprost, or travoprost monotherapy. Data collected from the HealthCore Integrated Research Database, as part of the Glaucoma Adherence and Persistency Study (GAPS), were used to populate the model. Patients with a documented diagnosis of glaucoma who were newly treated (no ocular hypotensive medication and no glaucoma-related procedure during 6 months before first prescription) with latanoprost, bimatoprost, or travoprost monotherapy were identified. The time horizon for the base-case model was the duration of chart abstraction (mean = 4.1 years); a 3-month model also was developed. Physician-reported rates of hyperemia were obtained from chart reviews of 300 patients. Transition rates reflected events related to reports of hyperemia where a physician-driven change (switch or discontinuation) in therapy was documented. The per-patient direct cost (2008) due to hyperemia-driven change in therapy was calculated as the sum of the cost of the initial prescription plus the cost of the office visit where the patient was evaluated and the decision to change therapy was made. Costs were stratified by whether patients were hyperemia free or discontinued the initial therapy due to hyperemia. From the sample of 13,977 newly treated patients, 8,743 patients were started on a prostaglandin monotherapy only. Of these, 5,726 received latanoprost, 1,633 were treated with bimatoprost, and 1,384 received travoprost index monotherapy. Across all treatment groups, costs among hyperemia-free patients were US$73.67 versus US$140.02 for those who discontinued the initial prostaglandin due to hyperemia. Per-patient costs were lowest in the group treated initially with latanoprost. For the base-case model, with latanoprost as the reference, total per-patient incremental costs due to hyperemia-driven change in therapy were US$5.92 for bimatoprost and US$5.43 for travoprost. Results were not highly sensitive to increases either in the incidence of hyperemia among latanoprost-treated patients or in the cost of latanoprost. Hyperemia results in increased overall costs in patients treated with latanoprost, bimatoprost, and travoprost. Treatment with latanoprost is associated with lower hyperemia-related costs than treatment with bimatoprost or travoprost.


Application of a synthetic aperture optical system to infrared imaging.
This paper discusses the application of a synthetic aperture system (SAS) for the formation of infrared, images. The SAS of primary interest is a hexagonal array of six circular apertures; special emphasis is placed on a one-dimensional model that describes the salient features of the two-dimensional model. In infrared images, in the 8-14-microm region, conventional detectors are too large to sample the image without aliasing the spatial spectrum. The unusual transfer function characteristics of the SAS are shown to obtain larger sampling distances than those required by the Whittaker-Shannon sampling theory. One possible detector array that will correctly sample the image is presented.


Rotary's PolioPlus Program: Lessons Learned, Transition Planning, and Legacy.
Hundreds of thousands of Rotary volunteers have provided support for polio eradication activities and continue to this day by making financial contributions to the Rotary PolioPlus program, participating in national immunization days, assisting with surveillance, working on local, national, and international advocacy programs for polio eradication, assisting at immunization posts and clinics, and mobilizing their communities for immunization activities (including poliovirus and other vaccines) and other health benefits. Rotary has contributed more than $1.61 billion for the global eradication of polio and has committed to provide an additional $35 million each year until 2018 (all dollar amounts represent US dollars). Its unwavering commitment to eradicate polio has been vital to the success of the program. Rotary is providing additional support for routine immunization and healthcare. When polio is finally gone, we will have the knowledge from the lessons learned with PolioPlus, such as the value of direct involvement by local Rotarians, the program for emergency funding, innovative tactics, and additional approaches for tackling other global issues, even those beyond public health. Rotary has already transitioned its grants program to include 6 areas of focus: disease prevention and treatment, water and sanitation, maternal and child health, basic education and literacy, economic and community development, and peace and conflict prevention/resolution. Funding for these grants in 2015-2016 was $71 million. The legacy of the polio program will be the complete eradication of poliovirus and the elimination of polio for all time.


[Primary culture and identification of mouse odontoblast-like cells].
To culture primary mouse odontoblast and to provide a base for study on inducing ES cells to odontoblast. Lower incisor germs were removed from 1-week-old mouse. The dental papillae were isolated in microscope, and the dental papillae cells were dispersed using 0.25% collagenase and 0.25% trypsin. The cell clones, which had similar morphology to odontoblasts, were selected for further culturing. The primary cultured cells were identified by light and electron microscopes and mRNA expression of mouse dentin sialophoprotein. The cultured cells had the same morphology and ultrastructure. They were rich in Golgi's complex, ribosome and rough endoplasmic reticulum. These cells expressed DSPP at mRNA levels. The cultured cells were mouse odontoblast-like cells. The method could be used for the study of odontal cells in vitro.


Pathogens implicated in acute otitis media failures after 7-valent pneumococcal conjugate vaccine implementation in France: distribution, serotypes, and resistance levels.
Before 7-valent pneumococcal conjugate vaccine (PCV7) implementation in France, several studies had described the microbiology of acute otitis media (AOM) treatment failures. The causative pathogens were Streptococcus pneumoniae (Sp) followed by nontypable Haemophilus influenzae (NTHi). The aim of this study was to describe the epidemiology of pathogens involved in AOM treatment failures or recurrences. This French multicentric prospective study enrolled 143 children with AOM treatment failure between 2007 and 2009 observed by 8 ear, nose, and throat specialists. Failure was defined as the persistence of AOM symptoms after at least 48 hours of antibiotic therapy or their recurrence within 4 days after the end of treatment. Standardized history and physical examination findings were recorded, and culture of middle ear fluid (MEF) was obtained. Mean age was 16.9 ± 9.9 months (median, 13.7). Eighty-eight percent of children had received more than 1 dose of PCV7, and 70.6% attended day care. The most common antibiotic used at the time of treatment failure or recurrence was a combination of amoxicillin and clavulanate (51.1%). Bacteriologic sampling demonstrated that in 35% of cases (n=50), no otopathogen was cultured at the time of treatment failure or recurrence. Similar proportions of Sp and NTHi were observed in the 86 patients (60.1%) from whom only a single species was recovered from MEF (46.5% for Sp, n=40 and 45.3% for NTHi, n=39). Among Sp strains, 4.4% were penicillin susceptible, 77.8% were penicillin intermediate, and 17.8% were fully penicillin resistant, and serotype 19A represented 84.5% of all serotypes detected. Among NTHi isolates, 15.5% (n=7) were β-lactamase-producing strains (including 2 strains with only this mechanism of resistance), and strains with reduced susceptibility by changes in protein binding to penicillin (β-lactamase-negative ampicillin resistant strains) represented 35.5% of cases. Among the 50 sterile MEF samples, polymerase chain reaction was performed in 32, of which 4 were positive for HI, 3 for Sp, and 3 for both. Among children with AOM treatment failures in France, Sp and NTHi were equally distributed; 19A was the main Sp serotype, and the main resistance mechanism for NTHi was β-lactamase-negative ampicillin resistance.


Structural characterization of polysaccharide-coated iron oxide nanoparticles produced by Staphylococcus warneri, isolated from a thermal spring.
The biocompatible-coated iron oxide nanoparticles (IONs) have attracted a great interest because of their various applications in biological science and medicine. In most cases, the toxic effect of naked iron oxide nanoparticles is completely cleared by adding a biocompatible coating, such as polysaccharides, polyethylene glycol (PEG), or biosynthesis of biocompatible-coated IONs using microorganisms such as bacteria. In the present study, polysaccharide-coated iron oxide nanoparticles were produced by a strain of Staphylococcus warneri isolated from a thermal spring. For identification of the isolated bacterium, 16S rRNA gene sequencing was done. Characterization of the nanoparticles was performed for the first time, using transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA), X-ray crystallography (XRD), Fourier-transform infrared (FTIR) spectroscopy, vibrating sample magnetometer (VSM), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results indicated that the spherical iron oxide nanoparticles were coated by a polysaccharide (13.6%), which provided a large negative charge of -91 mV and very low saturation magnetization of around 0.28 emu/g. The result of MTT assay on MOLT-4 cell lines showed that the percentage of viability was between 95.6% and 68.9% in the 10-100 µM of nanoparticle concentrations with a high IC 50 value, which makes it appropriate for biomedical applications such as cancer therapy.


Measurement properties and performance of an eight-minute submaximal treadmill test in patients with juvenile idiopathic arthritis: a controlled study.
Poor cardiorespiratory fitness is previously reported in patients with juvenile idiopathic arthritis (JIA) measured both by maximal and submaximal exercise tests, but a submaximal exercise test with acceptable measurement properties is currently lacking for both clinical and research purposes in this patient population. The objectives of this study were to evaluate the measurement properties and performance of a submaximal treadmill test in patients with JIA, and to compare the results with those obtained in controls. Fifty-nine patients (50 girls), aged 10-16 years, with oligo- (n = 30) and polyarticular (n = 29) JIA, and 59 age- and sex-matched controls performed an eight-minute submaximal treadmill test for estimating peak oxygen uptake (VO2peak) followed by a maximal treadmill test measuring VO2peak directly. During the submaximal treadmill test, the study participants walked with no inclination at a speed between 3.2-7.2 km/h for four minutes, and then continued to walk at the same speed for four minutes with five % inclination. VO2peak was directly measured during a continuous graded exercise test on treadmill until exhaustion. Thirty-seven patients participated in the evaluation of the reliability. Criterion validity and reliability were evaluated with interclass correlation coefficient (ICC); measurement errors by Bland-Altman plot, standard error of measurement and smallest detectable change. In patients with JIA, the ICC (95% CI) for criterion validity was acceptable at group level 0.71 (0.51, 0.82), but not at individual level. The test-retest reliability and inter-rater reliability were acceptable at individual (0.84 (0.71, 0.91) and 0.92 (0.83, 0.96), respectively) and group levels (0.91 (0.83, 0.96) and 0.96 (0.91, 0.98), respectively). The measurement errors (for test-retest reliability/inter-rater reliability) were large. Bland-Altman plots showed no systematic differences, but a large variability for both the validity and reliability. The performance of and estimated VO2peak from the submaximal test were not associated with disease variables and were comparable between patients and controls. The submaximal treadmill test is valid for use in patients with JIA on group level, but not on individual level. The reliability is acceptable. Due to large measurement errors, the submaximal treadmill test is not optimal for use in daily clinical practice to estimate VO2peak in individual patients.


Modified PVA-Fe3O4 nanoparticles as protein carriers into sperm cells.
Magnetite nanoparticles conjugated to protein are developed in order to potentially serve as protein carriers into bovine sperm cells. The conjugate comprises iron oxide nanoparticles that are covalently bound to an anti-protein kinase C (PKC)alpha antibody. This conjugate can serve for cellular PKC localization and the inhibition of its function. The surface of the nanoparticle is first modified with (3-aminopropyl) thrimethoxysilane to form a self-assembled monolayer, and subsequently conjugated with the antibody through amidation between the carboxylic acid end groups on the antibody and the amine groups on the surface of the nanoparticles. The anti-PKCalpha localization is proven by fluorescent microscopy and iron staining. The activity of the anti-PKCalpha conjugated with the nanoparticle is tested by recognizing PKCalpha using the Western blot method.


Office lab equipment. Aids in selection and cost analysis.
Factors determining the setup of your office laboratory include your anticipated benefits, such as rapid turnaround time, ease of operation, accuracy, convenience to patients, and profit; the type and volume of tests done in your office; percentage of reimbursement received from third-party payers; office space currently available for a lab; additional training required for office personnel; and instrument selection. As a aid in determining cost analysis of necessary equipment, the author provides work sheets for evaluation of various brands of instruments, calculation of your break-even point following purchase of equipment, and revenue analysis.


Characterization of dental pulp defect and repair in a canine model.
To explore a relationship between the size of pulp chamber perforation and reparative dentin formation in a canine model. Pulp defects were created in the pulp chambers of maxillary and mandibular premolars (N = 64) in 17 healthy mongrel dogs in three different sizes (diameter/depth: 1/1, 2/1, and 2/2 mm3) with sterile round burs under general anesthesia. The perforations were immediately capped with hard-setting calcium hydroxide (CH) in the control group or sealed with Teflon membrane (TM) in the experimental group, followed by restoration with reinforced zinc oxide eugenol cement in vivo. Seven and 30 days after pulp chamber perforation and restoration all treated and control premolars were extracted and prepared for histomorphometric and statistical analyses. Reparative dentin formation was more pronounced for defect sizes up to 2/1 mm3 when treated with CH, and completely bridged the surgically created dentin defects only after 30 days. However, reparative dentin upon CH treatment failed to completely bridge pulp chamber exposure for 2/2 defects. By contrast, TM treatment only yielded mild reparative dentin bridging for defects up to 1/1, but not for either 2/1 or 2/2 defects at 30 days. Inflammatory responses of the exposed dental pulp tissue were more robust with the TM group than with the CH group. Thus, dental pulp tissue possesses a capacity for spontaneous repair by the formation of reparative dentin in this preclinical model, but only up to a defect size of -2 mm in diameter and 1 mm in depth. All observations are based on 30 days post-treatment in the canine model. These findings may serve as baseline for regenerative endodontic studies.


Dicrocoelium dendriticum in cattle from Cypress Hills, Canada: humoral response and preliminary evaluation of an ELISA.
Apparent changes in the distribution of Dicrocoelium dendriticum in western Canada and the absence of other liver flukes in the area prompted an examination of the humoral responses in naturally infected cattle sharing pasture with heavily infected wild cervids. Western blots using sera from the infected cattle indicated that the cattle had antibodies to various fluke antigens, however the pattern of response varied among isotypes. Isotype analysis indicated that Dicrocoelium-infected cattle produced IgG1, and IgM antibodies, but not IgG2 antibodies. Subsequent comparison of the efficiency of coprological and serological approaches for diagnosis showed that while 43.5% of a separate group of 26 yearlings had eggs in their faeces, the immunoblots indicated that all had detectable antibodies. A group of cattle necropsied for confirmation of fluke status was used to evaluate humoral immune responses and to provide baseline information in the preliminary development of an indirect ELISA for detection of anti-fluke antibodies.