Determinants of left ventricular mass in normotensive children.
Left ventricular hypertrophy has been shown to be an independent predictor of cardiovascular morbidity and mortality. Few studies have examined the determinants of left ventricular mass (LVM) in normotensive children. Eight-four healthy children, blacks and whites, girls and boys, between 6 and 18 years of age, all with positive family histories of essential hypertension, participated in the study. Demographic and anthropometric characteristics, physical activity, and hemodynamic responses at rest and in response to the stress of forehead cold stimulation and a challenging video game were related to M-mode echocardiographic determined LVM indexed by two indices of body habitus. Hierarchical stepwise multiple regression analysis indicated that the significant independent correlates of LVM per body surface area were gender (boys were greater), baseline systolic blood pressure and heart rate, a physical activity index (sweat episodes per week) and peak systolic blood pressure responses to the forehead cold stressor (final model r2 = 0.49). For LVM/height2.7 significant independent correlates were adiposity index (weight z score--height z score), gender (boys were greater), age, resting systolic pressure, and systemic vascular resistance reactivity to forehead cold stimulation (final model r2 = 0.42). These findings with normotensive children corroborate other findings that have typically involved hypertensive children, indicating that resting heart rate and systolic pressure, gender, and adiposity are early determinants of LVM indices in children. In addition, the current findings indicate that hemodynamic responses to stress also appear to play a role in the early development of LVM.(ABSTRACT TRUNCATED AT 250 WORDS)


Liver Metabolomics Analysis Revealing Key Metabolites Associated with Different Stages of Nonalcoholic Fatty Liver Disease in Hamsters.
Nonalcoholic fatty liver disease (NAFLD) is not only the top cause of liver diseases but also a hepatic-correlated metabolic syndrome. This study performed untargeted metabolomics analysis of NAFLD hamsters to identify the key metabolites to discriminate different stages of NAFLD. Hamsters were fed a high-fat diet (HFD) to establish the NAFLD model with different stages (six weeks named as the NAFLD1 group and twelve weeks as the NAFLD2 group, respectively). Those liver samples were analyzed by untargeted metabolomics (UM) analysis to investigate metabolic changes and metabolites to discriminate different stages of NAFLD. The significant liver weight gain in NAFLD hamsters was observed, accompanied by significantly increased levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Moreover, the levels of TG, LDL-C, ALT, and AST were significantly higher in the NAFLD2 group than in the NAFLD1 group. The UM analysis also revealed the metabolic changes; 27 differently expressed metabolites were detected between the NAFLD2 and NAFLD1 groups. More importantly, the levels of N-methylalanine, allantoin, glucose, and glutamylvaline were found to be significantly different between any two groups (control, NAFLD2 and NAFLD1). Receiver operating characteristic curve (ROC) curve results also showed that these four metabolites are able to distinguish control, NAFLD1 and NAFLD2 groups. This study indicated that the process of NAFLD in hamsters is accompanied by different metabolite changes, and these key differently expressed metabolites may be valuable diagnostic biomarkers and responses to therapeutic interventions.


Dissecting the total transition state stabilization provided by amino acid side chains at orotidine 5'-monophosphate decarboxylase: a two-part substrate approach.
Kinetic analysis of decarboxylation catalyzed by S154A, Q215A, and S154A/Q215A mutant yeast orotidine 5'-monophosphate decarboxylases with orotidine 5'-monophosphate (OMP) and with a truncated nucleoside substrate (EO) activated by phosphite dianion shows (1) the side chain of Ser-154 stabilizes the transition state through interactions with the pyrimidine rings of OMP or EO, (2) the side chain of Gln-215 interacts with the phosphodianion group of OMP or with phosphite dianion, and (3) the interloop hydrogen bond between the side chains of Ser-154 and Gln-215 orients the amide side chain of Gln-215 to interact with the phosphodianion group of OMP or with phosphite dianion.


Redescription of two Dicyemennea (Phylum: Dicyemida) from Rossia pacifica (Mollusca: Cephalopoda: Decapoda).
Two species of dicyemid mesozoan are redescribed from Rossia pacifica Berry, 1911, collected off Iwase in Toyama Bay, Honshu, Japan. Dicyemennea brevicephaloides Bogolepova-Dobrokhotova, 1962, is a large species that reaches about 4,000 microm in length. This species attaches to the surface of the branchial heart appendages. The vermiform stages are characterized as having 23 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Infusoriform embryos consist of 35 cells; a single nucleus is present in each urn cell and the refringent bodies are absent. Dicyemennea rossiae Bogolepova-Dobrokhotova, 1962, is a medium species that reaches about 2000 microm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 29-34 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid.


Thaumatin and gum arabic allergy in chewing gum factory workers.
Thaumatin is a sweetener and flavor modifier commonly used in the food industry. Likewise, gum arabic is widely used as a food stabilizer and thickening agent. We report here that a powder mixture composed of 10% thaumatin and 90% gum arabic led to allergic symptoms in the upper airways in occupationally exposed individuals: four of eight workers of a chewing gum factory exposed to this powder mixture had pronounced rhinitis. A positive skin prick test result for pure thaumatin was obtained in all four individuals with rhinitis of whom two also had a positive skin prick test result for pure gum arabic and gum arabic-specific IgE. Subsitution of a powdered thaumatin with a liquid form reduced symptoms among the rhinitic workers. Although gum arabic is a well-known potential allergen, we were unable to find prior documentation of allergic symptoms to thaumatin when it is used in the food industry.


Perioperative care of infants with pyloric stenosis.
Pyloric stenosis (PS) is one of the most common surgical conditions affecting neonates and young infants. The definitive treatment for PS is surgical pyloromyotomy, either open or laparoscopic. However, surgical intervention should never be considered urgent or emergent. More importantly, emergent medical intervention may be required to correct intravascular volume depletion and electrolyte disturbances. Given advancements in surgical and perioperative care, morbidity and mortality from PS should be limited. However, either may occur related to poor preoperative resuscitation, anesthetic management difficulties, or postoperative complications. The following manuscript reviews the current evidence-based medicine regarding the perioperative care of infants with PS with focus on the preoperative assessment and correction of metabolic abnormalities, intraoperative care including airway management (particularly debate related to rapid sequence intubation), maintenance anesthetic techniques, and techniques for postoperative pain management. Additionally, reports of applications of regional anesthesia for either postoperative pain control or as an alternative to general anesthesia are discussed. Management recommendations are provided whenever possible.


The art of providing resuscitation in Greek mythology.
We reviewed Greek mythology to accumulate tales of resuscitation and we explored whether these tales could be viewed as indirect evidence that ancient Greeks considered resuscitation strategies similar to those currently used. Three compendia of Greek mythology: The Routledge Handbook of Greek Mythology, The Greek Myths by Robert Graves, and Greek Mythology by Ioannis Kakridis were used to find potentially relevant narratives. Thirteen myths that may suggest resuscitation (including 1 case of autoresuscitation) were identified. Methods to attempt mythological resuscitation included use of hands (which may correlate with basic life support procedures), a kiss on the mouth (similar to mouth-to-mouth resuscitation), application of burning torches (which might recall contemporary use of external defibrillators), and administration of drugs (a possible analogy to advanced life support procedures). A careful assessment of relevant myths demonstrated that interpretations other than medical might be more credible. Although several narratives of Greek mythology might suggest modern resuscitation techniques, they do not clearly indicate that ancient Greeks presaged scientific methods of resuscitation. Nevertheless, these elegant tales reflect humankind's optimism that a dying human might be restored to life if the appropriate procedures were implemented. Without this optimism, scientific improvement in the field of resuscitation might not have been achieved.


Pancreatitis-Induced Depletion of Syntaxin 2 Promotes Autophagy and Increases Basolateral Exocytosis.
Pancreatic acinar cells are polarized epithelial cells that store enzymes required for digestion as inactive zymogens, tightly packed at the cell apex. Stimulation of acinar cells causes the zymogen granules to fuse with the apical membrane, and the cells undergo exocytosis to release proteases into the intestinal lumen. Autophagy maintains homeostasis of pancreatic acini. Syntaxin 2 (STX2), an abundant soluble N-ethyl maleimide sensitive factor attachment protein receptor in pancreatic acini, has been reported to mediate apical exocytosis. Using human pancreatic tissues and STX2-knockout (KO) mice, we investigated the functions of STX2 in zymogen granule-mediated exocytosis and autophagy. We obtained pancreatic tissues from 5 patients undergoing surgery for pancreatic cancer and prepared 80-μm slices; tissues were exposed to supramaximal cholecystokinin octapeptide (CCK-8) or ethanol and a low concentration of CCK-8 and analyzed by immunoblot and immunofluorescence analyses. STX2-KO mice and syntaxin 2+/+ C57BL6 mice (controls) were given intraperitoneal injections of supramaximal caerulein (a CCK-8 analogue) or fed ethanol and then given a low dose of caerulein to induce acute pancreatitis, or saline (controls); pancreata were isolated and analyzed by histology and immunohistochemistry. Acini were isolated from mice, incubated with CCK-8, and analyzed by immunofluorescence microscopy or used in immunoprecipitation experiments. Exocytosis was quantified using live-cell exocytosis and Ca2+ imaging analyses and based on formation of exocytotic soluble N-ethyl maleimide sensitive factor attachment protein receptor complexes. Dysregulations in autophagy were identified using markers, electron and immunofluorescence microscopy, and protease activation assays. Human pancreatic tissues and dispersed pancreatic acini from control mice exposed to CCK-8 or ethanol plus CCK-8 were depleted of STX2. STX2-KO developed more severe pancreatitis after administration of supramaximal caerulein or a 6-week ethanol diet compared with control. Acini from STX2-KO mice had increased apical exocytosis after exposure to CCK-8, as well as increased basolateral exocytosis, which led to ectopic release of proteases. These increases in apical and basolateral exocytosis required increased formation of fusogenic soluble N-ethyl maleimide sensitive factor attachment protein receptor complexes, mediated by STX3 and STX4. STX2 bound ATG16L1 and prevented it from binding clathrin. Deletion of STX2 from acini increased binding of AT16L1 to clathrin, increasing formation of pre-autophagosomes and inducing autophagy. Induction of autophagy promoted the CCK-8-induced increase in autolysosome formation and the activation of trypsinogen. In studies of human pancreatic tissues and pancreata from STX2-KO and control mice, we found STX2 to block STX3- and STX4-mediated fusion of zymogen granules with the plasma membrane and exocytosis and prevent binding of ATG16L1 to clathrin, which contributes to induction of autophagy. Exposure of pancreatic tissues to CCK-8 or ethanol depletes acinar cells of STX2, increasing basolateral exocytosis and promoting autophagy induction, leading to activation of trypsinogen.


Naturopathic Oncology Care for Thoracic Cancers: A Practice Survey.
There is a lack of information on therapies recommended by naturopathic doctors (NDs) for lung and gastroesophageal cancer care. Study objectives were to: (1) identify the most common interventions considered for use by NDs; (2) identify interventions NDs recommend to support key therapeutic goals; and (3) identify potential contraindications between integrative and conventional therapies. Oncology Association of Naturopathic Physicians (OncANP) members (n = 351) were invited to complete an electronic survey. Respondents provided information on interventions considered for thoracic cancer pre- and postoperatively across 4 therapeutic domains (supplemental natural health products, physical, mental/emotional, and nutritional), therapeutic goals, and contraindications. This survey was part of the development of the Thoracic Perioperative Integrative Surgical Evaluation trial. Forty-four NDs completed the survey (12.5% response rate), all of whom were trained at accredited colleges in North America and the majority of whom were Fellows of the American Board of Naturopathic Oncology (FABNO) (56.8%). NDs identified significantly more interventions in the postoperative compared to preoperative setting. The most frequently identified interventions included modified citrus pectin, arnica, omega-3 fatty acids, vitamin D, probiotics, exercise, acupuncture, meditation, stress reduction, low glycemic index diet, and Mediterranean diet. Potential contraindications with conventional treatment (surgery, chemotherapy, radiotherapy) differed across natural health products. These findings highlight naturopathic interventions with a high level of use in thoracic cancer care, describe and characterize therapeutic goals and the interventions used to achieve these goals, and provide insight on how practice changes relative to conventional cancer treatment phase.


Cortical representation of the urge to void: a functional magnetic resonance imaging study.
The urge to void generally increases with bladder distention but the relationship between the 2 factors is complex. When the bladder is moderately filled, the desire to void can be called forth deliberately but it can also be suppressed. To elucidate human brain mechanisms that are active during such intentional modulations of the desire to void we performed functional magnetic resonance imaging in healthy volunteers. Brain activity was studied in 22 young women. At moderate bladder filling (about 350 ml) they periodically suppressed or enhanced the urge to void without allowing urine to pass. A manual task with a dynamometer, in which the current urge to void was expressed as grip force, demonstrated that the intensity of sensations could be influenced voluntarily. We also examined brain activity during repetitive (1 Hz) contractions of pelvic floor muscles after the bladder was emptied. Significant brain activity associated with an increased urge to void was found in the insular cortex, frontal opercula, supplementary motor area (SMA), cingulate motor area (CMA), right posterior parietal cortex, left prefrontal cortex and cerebellum. Trends toward activation were detected in the thalamus, peri-aquaeductal gray matter and ventral pons. Suppression of the urge to void significantly activated the left superior frontal lobe. The SMA and CMA were active during voluntary rhythmical contractions of pelvic floor muscles. Sensation intensity of the desire to void can be influenced intentionally. Frontoparietal cortical areas and the SMA/CMA seem to be involved in this process.


Bilateral congenital anomalies of the extracranial vertebral artery: management with balloon therapy.
Congenital anomalies of the vertebral artery, including aneurysms and fenestrations, or partial duplication of the extracranial vertebral artery are rarely seen. Presented here is an unusual case of bilateral congenital anomalies of the extracranial vertebral arteries. The right vertebral artery demonstrated an aneurysm and the left a partial duplication. The aneurysm was felt to be the source of the embolic phenomenon to the posterior circulation. This was treated with detachable balloon catheter placement.


Positron emission tomography-computerized tomography findings in a urinary bladder paraganglioma.
We present the staging fluorodeoxyglucose (FDG) positron emission tomography (PET-CT) findings in a patient with a functional urinary bladder paraganglioma. The PET-CT scan showed markedly increased FDG uptake into bilateral pelvic nodes that was consistent with regional nodal involvement. These findings were confirmed on histopathology. At present, there are no reports of PET-CT findings in urinary bladder paragangliomas.


[Comparison between immunofluorescence and immunoenzymatic assay for the rapid diagnosis of the respiratory syncytial virus in nasopharyngeal secretions].
An enzyme immunoassay, RSV-EIA Abbot, was evaluated by comparison with indirect immunofluorescence. Nasopharyngeal secretions obtained from 95 infants and young children with acute respiratory infections were examined for the presence of respiratory syncytial virus antigens with both methods. Specimens were stored at -70 degrees C before being tested by EIA. Out of 60 samples positive by indirect immunofluorescence, 46 were also positive by RSV-EIA (sensitivity 78.7%) and 34 out of 35 immunofluorescence negative specimens were negative by RSV-EIA (specificity 97.1%). Therefore, the EIA appears to be an acceptable test for the rapid detection of RSV as an alternative for indirect immunofluorescence.


Effects of progesterone on concentrations of monoamines in hypothalamic areas and plasma prolactin levels in rats.
The role of progesterone to increase prolactin (PRL) secretion on the first estrous day in pubertal rats was compared with its role in adult cyclic rats. The first estrus was induced by the administration of pregnant mare serum gonadotrophin (5 IU) at 28 days of age. A subcutaneous administration of 2.5 or 7.5 mg of progesterone/100 g body wt significantly increased the concentration of plasma PRL in pubertal rats within 4 hr. The PRL level obtained after progesterone administration was greater than that in similarly treated adult rats. The concentration of dopamine in the arcuate nucleus-median eminence (ARC-ME) in pubertal rats significantly decreased after a lower dosage of progesterone was administered, but no change was found in the preoptic area concentration. In adult estrous rats, the concentration of dopamine in the ARC-ME showed a tendency to decrease after the administration of a larger dose of progesterone (7.5 mg/100 g body wt). No change was observed in the concentrations of indoleamines in the preoptic area and ARC-ME after the administration of progesterone in both pubertal and adult rats. The concentrations of dopamine in the preoptic area and ARC-ME were lower in pubertal rats than in adults. The concentration of 5-hydroxyindole acetic acid and the ratio of 5-hydroxyindole acetic acid to 5-hydroxytryptamine in the ARC-ME were higher in pubertal rats than in adults. These results indicate that progesterone causes a greater increase in tonic PRL secretion in pubertal rats than in adult rats and that a lower hypothalamic dopamine activity and a higher serotonin activity in pubertal rats may account for these differences.


Effect of baicalein on apoptosis of the human Hep G2 cell line was induced by mitochondrial dysfunction.
The effects of baicalein on the human hepatoblastoma G2 (Hep G2) cell line were investigated in this study. By an SRB viability assay, we demonstrated that baicalein reduced the viability in a dose- and time-dependent manner. The apoptotic features such as chromatin condensation and DNA fragmentation were observed in the baicalein-treated cells. During the process of apoptosis, we noticed a sequential dissipation of mitochondrial membrane potential (DeltaPsim) and an apparent redistribution of cytochrome c from the mitochondria to the cytosol in baicalein-treated cells. Furthermore, the mitochondrial Bcl-2 protein represented a dramatic change in response to baicalein treatment. Altogether, our data suggested that the effect of baicalein on apoptosis of the human Hep G2 cell line was induced by mitochondrial dysfunction and Bcl-2 regulation.


New group of xylene linker-containing acetylcholinesterase reactivators as antidotes against the nerve agent cyclosarin.
Nerve agents such as sarin, cyclosarin and tabun are organophosphorus substances able to inhibit the enzyme acetylcholinesterase (AChE; EC 22.214.171.124). AChE reactivators and anticholinergics are generally used as antidotes in the case of intoxication with these agents. None of the known AChE reactivators is able to reactivate AChE inhibited by all nerve agents used. In this work, reactivation potency of nine newly developed AChE reactivators with an incorporated xylene ring in their structure was measured in vitro. Cyclosarin was chosen as an appropriate member of the nerve agent family. Reactivation potency of the tested AChE reactivators was compared with the gold standard of AChE reactivators--pralidoxime. Two oximes (K107 and K108) surpassed the reactivation potency of pralidoxime. Moreover, from the obtained results it could be deduced that AChE reactivators with a functional oxime group in position-2 are the most potent AChE reactivators in the case of cyclosarin intoxications.


Modeling Ultrafast Electron Dynamics in Strong Magnetic Fields Using Real-Time Time-Dependent Electronic Structure Methods.
An implementation of real-time time-dependent Hartree-Fock (RT-TDHF) and current density functional theory (RT-TDCDFT) for molecules in strong uniform magnetic fields is presented. In contrast to earlier implementations, the present work enables the use of the RT-TDCDFT formalism, which explicitly includes field-dependent terms in the exchange-correlation functional. A range of current-dependent exchange-correlation functionals based on the TPSS functional are considered, including a range-separated variant, which is particularly suitable for application to excited state calculations. The performance of a wide range of propagator algorithms for real-time methods is investigated in this context. A recently proposed molecular orbital pair decomposition analysis allows for assignment of electronic transitions, providing detailed information about which molecular orbitals are involved in each excitation. The application of these methods is demonstrated for the electronic absorption spectra of N2 and H2O both in the absence and in the presence of a magnetic field. The dependence of electronic spectra on the magnetic field strength and its orientation relative to the molecule is studied. The complex evolution of the absorption spectra with magnetic field is rationalized using the molecular orbital pair decomposition analysis, which provides crucial insight in strong fields where the spectra are radically different from their zero-field counterparts.


Circulating FGF19 closely correlates with bile acid synthesis and cholestasis in patients with primary biliary cirrhosis.
Bile acid (BA) synthesis in the liver is regulated by Fibroblast Growth Factor 19 (FGF19) secreted from the ileum as an enterohepatic feedback mechanism. Although FGF19 mRNA is absent in normal liver, FGF19 gene expression was reported to increase in response to both extrahepatic and intrahepatic cholestasis. The impact of upregulated FGF19 expression on BA synthesis is unclear and the overall role of circulating FGF19 and BA synthesis under cholestatic conditions needs to be further investigated. BA synthesis was directly quantified by measuring serum concentrations of 7alpha-hydroxycholest-4-en-3-one (C4), along with serum FGF19 and other parameters, in 44 patients with primary biliary cirrhosis (PBC) and 10 healthy subjects. Serum C4 were substantially lower, while those of FGF19 were higher, in cirrhotic PBC patients, as compared to those of either healthy or non-cirrhotic PBC patients. Analyses of the relationships between circulating FGF19, BA synthesis and cholestasis revealed that circulating FGF19 was strongly correlated with BA synthesis (r = -0.735, p<0.0001) and the severity of cholestasis (r = 0.590, p<0.001). Moreover, BA synthesis was found to be strongly correlated with the degree of cholestasis (r = 0.522, p = 0.0005). These findings demonstrate that the regulation of BA synthesis in response to cholestasis is primarily controlled by circulating FGF19 and that under cholestatic conditions, the FGF19-BA synthesis feedback mechanism remains intact. Administering FGF19, or suitable mimetic, as a pharmacological intervention to increase circulating levels of FGF19 and suppress BA synthesis by inhibiting CYP7A1 gene expression is likely to provide therapeutic benefits for many PBC patients.


Surgical management of retinal detachment resulting from macular hole in a setting of high myopia.
To evaluate the anatomical and visual outcomes of pars plana vitrectomy (PPV) with internal limiting membrane (ILM) peeling and use of retinal tamponade for retinal detachments resulting from macular hole (MHRD) in highly myopic eyes. Twenty-nine highly myopic patients (29 eyes) underwent PPV with ILM peeling and retinal tamponade for MHRD were enrolled. Demographics and best-corrected visual acuity (BCVA) were measured preoperatively and at final follow-up. Anatomical success and macular hole closure were analyzed. Patients' mean age of patients was 58.7 ± 10.6 years, mean follow-up was 11.7 ± 7.4 months. Twenty three eyes (23/26, 88.5%) undergoing primary PPV combined with ILM peeling had successful initial retinal reattachment, including 19 eyes (19/19, 100%) with silicone oil tamponade and in 4 eyes (4/7, 57.1%) with sulfur hexafluoride (C3F8) tamponade. Overall anatomical success was achieved in 27 eyes (27/29, 93.1%). The macular hole closure was observed in 17 eyes (17/26, 65.4%) with final anatomical success. Compared to preoperative BCVA, the mean postoperative BCVA in the eyes with anatomical success was significantly improved (P = 0.007, Wilcoxon signed rank test). As a primary or secondary procedure, PPV combined with ILM peeling and usage of retinal tamponade serves as an effective method for MHRD in highly myopic eyes.


GC-binding factor 2 interacts with dishevelled and regulates Wnt signaling pathways in human carcinoma cell lines.
GC-binding factor 2 (GCF2), a transcriptional repressor that decreases the activity of several genes is capable of binding directly to the GC-rich sequence of the EGFR promoter and repressing the transcriptional activity of EGFR. In addition to its function as a transcriptional repressor, GCF2 can directly interact with other proteins such as flightless-1 (Fli-1). Many previous findings pertaining to the function of Fli-1 have suggested a role for fli-1 in providing a direct link between molecules involved in signal transduction pathways and the actin cytoskeleton. We hypothesized that GCF2, together with Fli-1, plays a role in regulating cytoskeleton function, cell migration, and/or morphology. In our study, we observed that GCF2 is crucial for the activation of RhoA, a small GTPase that plays a key role in the regulation of the actin cytoskeleton. RhoA was markedly inactivated as a result of the decreased expression of GCF2. Co-immunoprecipitations were subsequently performed to further investigate the mechanism for the repressive function. We identified dishevelled (Dvl), which is the key mediator for the Wnt pathway, as a binding partner with GCF2. These results strongly suggest that GCF2 plays a role in the Wnt-noncanonical planar cell polarity (PCP) signaling pathway. Consequently, GCF2 may regulate the cytoskeleton or migration via Dvls and RhoA.


Cross-Sphere Electrode Reaction: The Case of Hydroxyl Desorption during the Oxygen Reduction Reaction on Pt(111) in Alkaline Media.
Hydroxide ion is a common electrolyte when electrode reactions take place in alkaline media. In the case of oxygen reduction reaction on Pt(111), we demonstrate by ab initio molecular dynamics calculations that the desorption of hydroxyl (OH*) from the electrode surface to form a solvated OH- is a cross-sphere process, with the OH* reactant in the inner sphere and the OH- product directly generated in the aqueous outer sphere. Such a mechanism is distinct from the typical inner sphere and outer sphere reactions. It is dictated by the strong hydrogen bonding interactions between a hydroxide ion and water molecules and is facilitated by proton transfer through solvation layers. It should play a significant role whenever OH* desorption, or its reverse, OH- adsorption, is involved in an electrochemical reaction.


Richter's syndrome. A terminal complication of chronic lymphocytic leukemia with distinct clinicopathologic features.
Long, John C., AND Aisenberg, Alan C.: Richter's syndrome. A terminal complication of chronic lymphocytic leukemia with distinct clinicopathologic features. Am J Clin Pathol 63: 786-795, 1975. Two patients with chronic lymphocytic leukemia of 8 and 9 years duration had a terminal illness characterized by weight loss, persistent fever, lymphadenopathy, and dysglobulinemia. In both cases autopsy revealed a pleomorphic histiocytic lymphoma which contained multinucleate tumor cells and was associated with persistent chronic lymphocytic leukemia. These two cases are examples of Richter's syndrome, a clinically distinct complication of chronic lymphocytic leukemia that may be confused pathologically with Hodgkin's disease. Review of the medical literature suggests that this syndrome has frequently gone unrecognized. (Key words: Richter's syndrome; Chronic lymphocytic leukemia.)


[Antitumor effect of a triple negative breast cancer-dendritic cell fusion vaccine].
To test the antitumor effect of a human triple-negative breast cancer cell-dendritic cell (DC) fusion vaccine. DCs were isolated from fresh peripheral blood of healthy donors. The fusion vaccine was prepared by fusing the DCs and MDA-MB-231 cells via electrofusion. The morphology of the vaccine was identified under inverted fluorescence microscope and the phenotypes were analyzed with flow cytometry. The production of interleukin-12 (IL-12) and interferon-γ (IFN-γ) by the fusion cells was assessed using ELISA. A CCK-8 kit was used to examine the effect of the vaccine in stimulating the proliferation and cytotoxicity of autologous T lymphocytes. The DCs isolated from peripheral blood mononuclear cells highly expressed CD83, CD86, CD11c and HLA-DR on the cell surface. The fusion cells were irregular in shape and coexpressed the phenotypes of DCs and MDA-MB-231 cells. The fusion cells possessed a strong ability to stimulate the proliferation of T lymphocytes in vitro. Compared with the control group, the fusion vaccine showed a stronger antitumor effect against the breast cancer cells. The triple-negative breast cancer-DC fusion vaccine prepared by electrofusion can stimulate the proliferation of T lymphocytes and induces strong cytotoxicity of the T cells against breast cancer cells.


Effect of strong wind on laminas and petioles of Farfugium japonicum (L.) Kitam. var. japonicum (Asteraceae).
Farfugium japonicum (L.) Kitam. var. japonicum grows mainly in the coastal areas of Japan. Meteorological recording data from natural habitats were used to investigate the factors associated with the laminas and petioles of radical leaves of F. japonicum var. japonicum to avoid or resist higher wind stress. Our morphological and mechanical results indicated that petiole length and petiole cross-sectional area had a weak correlation with wind speed and breaking strength, and the petiole second area moment of inertia did not differ significantly among populations. However, both lamina area and petiole length per petiole cross-sectional area decreased with increasing wind speed, indicating that F. japonicum var. japonicum resisted or avoided an increase in wind speed outdoors by reducing the lamina area and petiole length per petiole cross-sectional area without qualitative changes in their petioles. The results of this study indicated that densely distributed recording stations of the Automated Meteorological Data Acquisition System (AMeDAS) by the Japan Meteorological Agency can be used for environmental adaptation studies of plants in the field using nearby plant populations.


Automatic location of vertebrae in digitized videofluoroscopic images of the lumbar spine.
Back pain is a widespread problem, and the disability it engenders continues to grow, despite efforts to contain it. A major problem in the diagnosis and management of back pain is the assessment of the degree to which mechanical factors play a part. Of considerable importance in understanding these mechanical factors is being able to quantify how the human spine actually moves in vivo. Digitized videofluoroscopy is currently the only practical method available for studying spinal motion in vivo at the segmental level. Low-dose, planar motion X-rays of the spine are captured on videotape and subsequently digitized for analysis. Until now, vertebrae in the digitized images were identified and marked manually as a basis for calculating intervertebral kinematics. This paper describes a procedure for automatically identifying the vertebrae in the motion sequences. The process increases objectivity and repeatability, and significantly reduces the manual effort required in locating the vertebrae prior to calculating the kinematics. The technique has been applied to images of a calibrated model and the results are promising. In-plane rotations may be calculated to an accuracy of at least 1 degree. Repeated analysis reveals standard deviations of less than 0.5 degree for intervertebral rotations and less than 0.25 mm for translations.


Biological significance of short-chain fatty acid metabolism by the intestinal microbiome.
Evidence suggests that short-chain fatty acids (SCFAs) derived from microbial metabolism in the gut play a central role in host homeostasis. The present review describes the current understanding and physiological implications of SCFAs derived from microbial metabolism of nondigestible carbohydrates. Recent studies indicate a role for SCFAs, in particular propionate and butyrate, in metabolic and inflammatory disorders such as obesity, diabetes and inflammatory bowel diseases, through the activation of specific G-protein-coupled receptors and modification of transcription factors. Established prebiotics, such as fructooligosaccharides and galactooligosaccharides, which support the growth of Bifidobacteria, mainly mediate acetate production. Thus, recent identification of prebiotics which are able to stimulate the production of propionate and butyrate by benign saccharolytic populations in the colon is of interest. Manipulation of saccharolytic fermentation by prebiotic substrates is beginning to provide information on structure-function relationships relating to the production of SCFAs, which have multiple roles in host homeostasis.


Drug Recognition Evaluation and Chemical Confirmation of a 25C-NBOMe-Impaired Driver.
This case report details an individual arrested for drug-impaired driving after leaving the scene of multiple motor vehicle collisions and evading police. The driver was examined by a drug recognition expert and failed the drug recognition evaluation. The driver admitted to using cocaine, marijuana, an antidepressant medication and "N-bomb," a novel psychoactive substance that possesses hallucinogenic properties. Toxicological analyses at the Centre of Forensic Sciences' Toronto laboratory revealed only the substance 2-[4-chloro-2,5-dimethoxyphenyl]-N-[(2-methoxyphenyl)methyl]ethanamine (25C-NBOMe) in the accused's urine. This is the first report in which 25C-NBOMe was identified through DRE and toxicological analyses in a drug-impaired driver.


Reductions in bacterial viability stimulate the production of Extra-intestinal Pathogenic Escherichia coli (ExPEC) cytoplasm-carrying Extracellular Vesicles (EVs).
Extra-intestinal Pathogenic Escherichia coli (ExPEC) is defined as an extra-intestinal foodborne pathogen, and several dominant sequence types (STs) ExPEC isolates are highly virulent, with zoonotic potential. Bacteria extracellular vesicles (EVs) carry specific subsets of molecular cargo, which affect various biological processes in bacteria and host. The mechanisms of EVs formation in ExPEC remains to be elucidated. Here, the purified EVs of ExPEC strains of different STs were isolated with ultracentrifugation processes. A comparative analysis of the strain proteomes showed that cytoplasmic proteins accounted for a relatively high proportion of the proteins among ExPEC EVs. The proportion of cytoplasm-carrying vesicles in ExPEC EVs was calculated with a simple green fluorescent protein (GFP) expression method. The RecA/LexA-dependent SOS response is a critical mediator of generation of cytoplasm-carrying EVs. The SOS response activates the expression of prophage-associated endolysins, Epel1, Epel2.1, and Epel2.2, which triggered cell lysis, increasing the production of ExPEC cytoplasm-carrying EVs. The repressor LexA controlled directly the expression of these endolysins by binding to the SOS boxes in the endolysin promoter regions. Reducing bacterial viability stimulated the production of ExPEC EVs, especially cytoplasm-carrying EVs. The imbalance in cell division caused by exposure to H2O2, the deletion of ftsK genes, or t6A synthesis defects activated the RecA/LexA-dependent SOS response, inducing the expression of endolysins, and thus increasing the proportion of cytoplasm-carrying EVs in the total ExPEC EVs. Antibiotics, which decreased bacterial viability, also increase the production of ExPEC cytoplasm-carrying EVs through the SOS response. Changes in the proportion of cytoplasm-carrying EVs affected the total DNA content of ExPEC EVs. When macrophages are exposed to a higher proportion of cytoplasm-carrying vesicles, ExPEC EVs were more cytotoxic to macrophages, accompanied with more-severe mitochondrial disruption and a higher level of induced intrinsic apoptosis. In summary, we offered comprehensive insight into the proteome analysis of ExPEC EVs. This study demonstrated the novel formation mechanisms of E. coli cytoplasm-carrying EVs.


Surgical management of lung metastases.
Data from 93 patients undergoing resection of lung metastases are examined. Survival at 5 years was 32% and operative mortality 2.1%. Histologically, metastases from carcinoma gave better results than metastases from sarcoma (5-year survival 40% and 0 respectively). The best overall results being obtained in the case of metastases from carcinoma of the testis and larynx (5-year survival 66% and 54% respectively). The free interval was shown to be a good therapeutic index when longer than 1 year (44% survival at 5 years when longer than 1 year, 16% when shorter). Resection of lung metastases is recommended when primary tumor is under control, there are no metastases to other organs, the operative risk is reasonable and no other effective therapeutic procedures are available.


Changes in standing body sway of pregnant women after long-term bed rest.
Pregnant women tend to fall and increased body postural instability, namely body sway, may be one of the causative factors. We had a clinical impression that pregnant women after long-term bed rest tend to fall. We hypothesised that such women may show increased body sway, which we attempted to determine. Pregnant women (n = 161) were divided into three groups: (i) women with preterm labour after 2-week bed rest, (ii) those after 4-week bed rest, and (iii) those without bed rest or preterm labour. Body sway was analysed using stabilometry, that is, computed analysis of movement of the centre of gravity. The 3 groups fundamentally showed the same stabilometric measurements. Women with oedema showed greater medial-lateral sway than those without it. Factors other than oedema yielded no differences in stabilometric parameters. Long-term bed rest fundamentally did not increase body sway to the extent that stabilometry could reveal it. It may be prudent to consider that pregnant women with oedema tend to fall.


Naturally-derived endoplasmic reticulum stress inhibitors for osteoarthritis?
Osteoarthritis (OA) is a chronic degenerative disease that affects the whole synovial joint. OA causes severe pain and disability that significantly affects the livelihood of an individual and incurs a huge socioeconomic burden. Current management strategies are limited to supporting functional improvement with physiotherapy and pain reduction as there are no drugs available that can reverse the progression of OA with only joint replacement surgery for late stage OA. OA is associated with advancing age and obesity, both of which compromise the functions of key endoplasmic reticulum (ER) molecular chaperones leading to improper protein folding and ER stress. Failure to restore protein homeostasis leads to increased cellular stress and eventually apoptotic cell death. Cartilage is avascular and is dependent on its constituent cells, chondrocytes, for extracellular matrix maintenance. Chondrocytes have limited proliferative capacity and their apoptosis eventually leads to extracellular matrix loss and cartilage degeneration. Recent studies on attenuating ER stress and chondrocytes apoptosis offer a credible strategy for reducing OA progression. The established roles of ER stress responses in OA have paved the way for targeted drug discovery studies aiming to mitigate ER stress and OA progression. In this review, in vitro, pre-clinical and clinical evidence of naturally-derived ER stress inhibitors for OA, the prospect and challenges in bringing these compounds to clinics are discussed.


Management of postmenopausal virilization.
Mild clinical signs of hyperandrogenism such as hirsutism may appear during the menopausal transition as part of the normal aging process, but the development of frank virilization suggests a specific source of androgen excess, including androgen-secreting tumors. A 68-yr-old postmenopausal woman was referred because of a history of progressive development of hirsutism and frontal balding for the previous 8 yr, together with moderate hyperandrogenemia. Initial imaging procedures depicted a 2-cm solid nodule in the right adrenal gland and normal appearance of both ovaries. To confirm the source of androgen excess, we conducted simultaneous selective venous sampling of adrenals and ovaries. Sampling was consistent with an ovarian source. After bilateral laparoscopic salpingo-oophorectomy, the patient was diagnosed with bilateral ovarian hyperthecosis. Three weeks after surgery, her androgen levels had decreased to the normal female range. Diagnosis of hyperandrogenism in postmenopausal women is challenging. Postmenopausal virilization may be associated with adrenal or ovarian androgen-secreting tumors or with benign conditions. A detailed clinical history is critical to differentiate the progressive development of virilization that characterizes benign causes from the rapid progression that characterizes malignant tumors. Imaging techniques do not always reveal the cause of hyperandrogenism and may even be misleading. Although technically difficult, combined adrenal and ovarian venous sampling may be required to confirm the source of androgen excess before the best surgical approach is determined.


Localization of immunoreactive testibumin in the testis and epididymis of adult rats.
Testibumin is a glycoprotein previously isolated from the spent media of primary Sertoli cell-enriched cultures prepared from 20-day-old rats. Immunoassayable testibumin is found in the highest concentrations in testis, epididymis and fluids of the male reproductive tract in adult rats. In the present study, light microscopy was used to show that immunostainable testibumin in paraffin sections of rat testis was localized along the base of the seminiferous epithelium and in finger-like projections from the base of the epithelium, corresponding to the position of the Sertoli cells. The immunostaining of Sertoli cells was shown to be specific since either purified testibumin or crude Sertoli cell-enriched culture medium could compete with antibody for binding sites in tissue sections. The observations using light microscopy were confirmed when Sertoli cells were examined by electron microscopy using a pre-embedding immunostaining technique. The epithelium of the epididymis also contained immunoreactive testibumin which was localized in the caput, corpus and cauda. Immunostainable testibumin was also localized in the corpora lutea of the rat ovary and in the epithelium of the uterine endometrium. These observations are consistent with previous reports that immunoreactive testibumin is present in these organs as demonstrated by radio-immunoassays. We conclude that (i) the immunolocalization of testibumin in Sertoli cells adds to a growing list of observations suggesting that it is made in this cell type; (ii) the other sites of testibumin synthesis in the male are uncertain but the ovary and uterus are possible sites in the female; (iii) electron microscopy following pre-embedding immunostaining and epitope selection can be used as an adjunct to conventional immunocytochemistry to localize proteins in Sertoli cells.


The selection and suppression of action: ERP correlates of executive control in humans.
A key aspect of higher cortical function is the ability to selectively withhold or suppress action where appropriate. To examine the time course of executive control we used dense-sensor EEG recording techniques to study event-related electrical potentials (ERPs) during a visual 'go/no-go' task. We show that during both go and no-go trials there is a positive deflection in the ERP, which develops over posterior parietal sensors approximately 350 ms (P300) after the onset of a conditional visual stimulus, but is selectively suppressed during no-go trials. We also show that this modulation of the parietal P300 is preceded by a negative deflection in the ERP recorded over frontal cortex (N2), which is apparent only for no-go trials. We suggest that this signal provides an electrophysiological marker in man for the decision to withhold the execution of a motor response.


Does social class predict substance problems in young adults with ADHD?
The relationship between social class and substance use problems is unclear. We aimed to clarify this association in a sample of young adult males with and without attention-deficit/hyperactivity disorder (ADHD). We included 69 ADHD and 78 control subjects. Substance use problems were measured with the Drug Use Screening Inventory-Revised (DUSI). Among ADHD subjects, we found a U-shaped association, with elevated risk for substance-related problems at both ends of the SES spectrum. No significant association was found in controls. These findings indicate that substance use risk in ADHD subjects is especially vulnerable to social class.


[The use of the "anti-G-suit" during operations in the sitting position as a treatment of postural hypotension.(author's transl)].
The use of the "Anti-G-Suit" in patients with postural hypotension is described. Acute falls in the blood pressure during operations in the sitting position can be successfully treated with the aid of an external counterpressure garment reaching from ankles to xiphoid process. The mean increase in systolic blood pressure was about 60% in hypotensive and 17% in normotensive control patients. A reciprocal relation between pre-treatment blood pressure and its increase after use of the "G-Suit" was noticed. In contrast to previous practice, we were able to operate on all hypotensive patients in poor condition. The effect of external counterpressure on the lower part of the body can be explained by increase of peripheral resistance and decrease of vascular flow, resulting in better venous return, displacement of the circulating blood above the diaphragm and augmentation of systolic blood pressure.


Tubal ectopic pregnancy: our experience from 2000 to 2013.
In this paper we summarize our experience in diagnosis and treatment of 402 retrospectively collected tubal EP and review the most recent topics from the literature. Systemic Methotrexate (MTX) was effective in 56 out of 65 patients (failure rate 13.8%), in whom hCG level was significantly lower when compared to the failure group (p<0,05); we performed 299 salpingectomies, 297 of whom through laparoscopic approach. MTX single-dose is safe and effective in eligible patients; surgery represents the treatment of most of the EPs, mainly through laparoscopic approach.


Environmental estrogens and reproductive biology in amphibians.
In this paper, the effects of an estrogenic compound, 4-nonyl-phenol (NP), on the amphibians Rana esculenta and Triturus carnifex are described together with those on sexual differentiation in Xenopus laevis. NP increased plasma vitellogenin in male frogs and newts in a dose-related manner; moreover, inhibitory effects on gonadotropin and prolactin (PRL) secretion by pituitary were found together with an elevation of plasma androgens. NP treatment also caused a remarkable increase in number of prolactin-immunolabeled cells, suggesting that xenoestrogen might induce, at least in the newt pituitary, a PRL accumulation possibly due to a reduction of the hormone release. In addition, both NP and bisphenol A caused feminization by increasing the percentage of female phenotypes in X. laevis, and the in vivo effects were more pronounced than those of estradiol-17beta.


Journey of patients with cancer: a systematic evaluation at tertiary care center in India.
In cancer patients early institution of therapy placed a very important role and delay in the diagnosis and treatment can cause catastrophe. Affirm step to cut shot this delay requires detailed information about each step of patient referral journey and for fulfillment of above aim, we interviewed 101 patients, to calculate the elapsed time at each step. Result revealed that onset of symptoms to median time of presentation to general practitioner is 20 (9 - 28) days, time consumed in state based hospital is 100 (15- 167) days while in Delhi based hospital is 56 (18 - 100) days. Higher cure rate (38.2%) in patients presented within 3 months of development of cancer symptoms than those presented late. Study concluded that primary physician and all the referral hospital attributed important role in early diagnosis and treatment of cancer.


Identification of Potential Bioactive Ingredients and Mechanisms of the Guanxin Suhe Pill on Angina Pectoris by Integrating Network Pharmacology and Molecular Docking.
The Guanxin Suhe pill (GSP), a traditional Chinese medicine, has been widely used to treat angina pectoris (AP) in Chinese clinical practice. However, research on the bioactive ingredients and underlying mechanisms of GSP in AP remains scarce. In this study, a system pharmacology approach integrating gastrointestinal absorption (GA) evaluation, drug-likeness (DL) evaluation, target exploration, protein-protein-interaction analysis, Gene Ontology (GO) enrichment analysis, network construction, and molecular docking was adopted to explore its potential mechanisms. A total of 481 ingredients from five herbs were collected, and 242 were qualified based on GA and DL evaluation. Target exploration identified 107 shared targets between GSP and AP. Protein-protein interaction identified VEGFA (vascular endothelial growth factor A), TNF (tumor necrosis factor), CCL2 (C-C motif chemokine ligand 2), FN1 (fibronectin 1), MMP9 (matrix metallopeptidase 9), PTGS2 (prostaglandin-endoperoxide synthase 2), IL10 (interleukin 10), CXCL8 (C-X-C motif chemokine ligand 8), IL6 (interleukin 6), and INS (insulin) as hub targets for GSP, which were involved in the inflammatory process, ECM proteolysis, glucose metabolism, and lipid metabolism. GO enrichment identified top pathways in the biological processes, molecular functions, and cell components, explaining GSP's potential AP treatment mechanism. Positive regulation of the nitric oxide biosynthetic process and the response to hypoxia ranked highest of the biological processes; core targets that GSP can regulate in these two pathways were PTGS2 and NOS2, respectively. Molecular docking verified the interactions between the core genes in the pathway and the active ingredients. The study lays a foundation for further experimental research and clinical application.


Imprinted nanoporous organosilicas for selective adsorption of nitroenergetic targets.
Periodic mesoporous organosilicas incorporating diethylbenzene bridges in their pore walls were applied for the adsorption of nitroenegetic targets from aqueous solution. The materials were synthesized by co-condensing 1,4-bis(trimethoxysilylethyl)benzene (DEB) with 1,2-bis(trimethoxysilyl)ethane to improve structural characteristics. Molecular imprinting of the pore surfaces was employed through the use of a novel target-like surfactant to further enhance selectivity for targets of interest (tri- and dinitrotoluenes) over targets of similar structure ( p-cresol and p-nitrophenol). The headgroup of the commonly used alkylene oxide surfactant Brij76 was modified by esterification with 3,5-dinitrobenzoyl chloride. This provided a target analogue which was readily miscible with the Brij76 surfactant micelles used to direct material mesopore structures. The impact of variations in precursor ratios and amounts of imprint molecule was evaluated. The use of 12.5% of the modified Brij surfactant with a co-condensate employing 30% DEB was found to provide the best compromise between total capacity and selectivity for nitroenergetic targets.


Competitive solid-phase immunoassay of testosterone using time-resolved fluorescence.
A competitive solid-phase immunoassay for the determination of testosterone in serum samples using time-resolved fluorescence is described. The solid phase is a testosterone-3-(O-carboxymethyl)-oxime-ovalbumin conjugate coated to polystyrene microtiter strips. Europium-labelled polyclonal and monoclonal antibodies against testosterone-3-(O-carboxymethyl)-oxime-bovine serum albumin were compared. Their behavior was quite similar although the polyclonal antibody was more sensitive, giving a detection limit of 15 fmol testosterone per assay. Correlation with RIA was very good (r = 0.982 and y = -0.150 + 0.969x).


Are dietary reports in a case-control study on thyroid cancer biased by risk perception of Chernobyl fallout?
In retrospective case-control studies performed following nuclear tests or nuclear accidents, individual thyroid radiation dose reconstructions are based on fallout and meteorological data from the residential area, demographic characteristics, and lifestyle as well as dietary information. Collecting the latter is a controversial step, as dietary declarations may be affected by the subjects' beliefs about their risk behavior. This report analyses the potential for such bias in a case-control study performed in eastern France. The study included 765 cases of differentiated thyroid carcinoma matched with 831 controls. Risk perceptions and beliefs of cases and controls were compared using Chi2 tests and differences in dietary reports were analyzed using a two-way ANOVA. In general, atmospheric pollution and living near a nuclear power plant were the two major risks that may influence thyroid cancer occurrence cited by cases and controls. When focusing in particular on the consequences of the Chernobyl accident, cases were more likely to think that the consequences were responsible for thyroid cancer occurrence than controls. Vegetable consumption during the two months after the Chernobyl accident was correlated with the status of subjects, but not to their beliefs. Conversely, consumption of fresh dairy products was not correlated with the status or beliefs of subjects. We found no evidence of systematic bias in dietary reports according to the status or beliefs held by subjects about the link between thyroid cancer occurrence and Chernobyl fallout. As such, these dietary reports may be used in further studies involving individual dosimetric reconstructions.


Severe Puumala virus infection in a patient with a lymphoproliferative disease treated with icatibant.
Early identification of patients at risk of a severe course of hantaviral disease and lack of effective medication represent a global challenge in the treatment of this emerging infection. We describe a 67-year-old female patient with a history of chronic lymphoproliferative disease involving the spleen and an extremely severe acute Puumala hantavirus infection. She was treated with the bradykinin receptor antagonist icatibant and recovered. She is the second patient with a spleen abnormality and severe Puumala infection treated with icatibant in our hospital. We suggest that patients with spleen abnormalities may be more susceptible to severe hantavirus disease. The activation of the kinin-kallikrein system and the formation of bradykinin in hantavirus-infected endothelial cells indicate that the role of bradykinin receptor antagonist icatibant in the treatment of hantavirus disease is worth studying.


Acute Coronary Syndrome Symptom Clusters: Illustration of Results Using Multiple Statistical Methods.
Researchers have employed various methods to identify symptom clusters in cardiovascular conditions, without identifying rationale. Here, we test clustering techniques and outcomes using a data set from patients with acute coronary syndrome. A total of 474 patients who presented to emergency departments in five United States regions were enrolled. Symptoms were assessed within 15 min of presentation using the validated 13-item ACS Symptom Checklist. Three variable-centered approaches resulted in four-factor solutions. Two of three person-centered approaches resulted in three-cluster solutions. K-means cluster analysis revealed a six-cluster solution but was reduced to three clusters following cluster plot analysis. The number of symptoms and patient characteristics varied within clusters. Based on our findings, we recommend using (a) a variable-centered approach if the research is exploratory, (b) a confirmatory factor analysis if there is a hypothesis about symptom clusters, and (c) a person-centered approach if the aim is to cluster symptoms by individual groups.


Plasmodium attenuation: connecting the dots between early immune responses and malaria disease severity.
Sterile attenuation of Plasmodium parasites at the liver-stage either by irradiation or genetic modification, or at the blood-stage by chemoprophylaxis, has been shown to induce immune responses that can protect against subsequent wild-type infection. However, following certain interventions, parasite attenuation can be incomplete or non-sterile. Instead parasites are rendered developmentally stunted but still capable of establishing an acute infection. In experiments involving Plasmodium berghei ANKA, a model of experimental cerebral malaria, it has been observed that several forms of attenuated parasites do not induce cerebral pathology. In this perspective we collect evidence from studies on murine malaria in particular, and attempt to "connect the dots" between early immune responses and protection from severe cerebral disease, highlighting potential parallels to human infection.


Studies of mitogen-induced lymphocyte transformation by a semi-microtechnic.
The authors have demonstrated that mitogen-induced lymphocyte transformation can be effectively studied in a semi-microsystem allowing multiple studies from a reasonably small volume of blood, which makes the technic more feasible for clinical studies of cellular immune mechanisms. The variability among individuals, and from day to day in the same individual, makes repeated, careful kinetic studies necessary for valid data. Moreover, the very wide range of normal responses in man makes the interpretation of cellular hyporesponsiveness difficult in pathologic states and necessitates the use of several mitogen concentrations and sequential kinetic studies to establish the validity of results.


Supramolecular Assembly and Microscopic Dynamics Modulation of Nanoscale Inorganic Cryptand and Polymer Complex for Versatile Design of Flexible Single-Ion Conductors.
The popular design of solid-state electrolytes (SSEs) from the chain relaxation of polymers faces the trade-offs among ion conductivity, stability, and processability. Herein, 2 nm inorganic cryptand molecules with the capability to carry different types of cations, including Ag+, Na+, K+, and Ca2+, are complexed with cationic polymers via ionic interaction, respectively, and the hybrid materials further phase separate into lamellar or hexagonal columnar structures. The successful establishment of ordered structures with ion channels from the packing of inorganic cryptands confers SSEs' excellent ionic conductivity to versatile types of cations. Meanwhile, suggested from the combination of broad dielectric spectroscopy, rheology, and thermal analysis, the fast chain relaxation can activate the dynamics of inorganic cryptand molecules and facilitate the ion hopping process in ion channels. The supramolecular interaction in the complex enables the highly flexible physical appearance for defect-free contact with electrodes as well as cost-effective processability and recyclability.


Longitudinal epidemiology of multiple sclerosis in Townsville, Queensland, Australia, 2012-2022.
Townsville (population=195,564, latitude=19.3°S) is the largest city in the Northern Queensland region of Australia, an area previously defined as a low/medium-prevalence zone for multiple sclerosis (MS). However, the epidemiology of MS in this region since 1981 is unknown. To assess the 2012 to 2022 epidemiology of MS in Townsville. Demographic/clinical data extracted from medical records of MS cases identified by public and private clinicians. Prevalence, and incidence and mortality rates estimated for 2012 and 2022 and age-standardised to the 2022 Australian population. Differences in estimates assessed by Poisson regression. Females and relapsing-remitting MS comprised most cases. The 2012 prevalence was 45.0/100,000 (50.4/100,000 age-standardised, F/M sex ratio=2.0). Prevalence increased by 188% in 2022, with a crude prevalence of 86.9/100,000 (91.7/100,000 age-standardised, F/M sex ratio=2.7). 2012-22 MS onset incidence rate was 3.8/100,000 person-years (age-standardised 3.5/100,000, F/M sex ratio=2.7). Mean age increased from 49.4 to 57.3 years. Age-standardised mortality rate was 0.9/100,000 person-years, with standardised mortality ratio=1.0. These results show that Townsville is a high-frequency region for MS, with prevalence and incidence on par with that seen at higher latitudes in Australia. These results have implications for clinical practice in the region and for organisational resource allocation.


Vitamin D: an old prohormone with an emergent role in chronic kidney disease.
The term vitamin D is linked to the development of rickets and osteomalacia, and has been known about since at least the 17th century. Prevention of these diseases has challenged many researchers in the following centuries, which has resulted in important advances in prevention and treatment. However, it was only in 1900 that the vitamin structure was identified. In the following decades, many steps were made toward understanding the metabolism of the vitamin. Recent progress has revolutionized our view of vitamin D and suggested that it has much wider effects on the body than ever believed. In addition to its well-known effects on calcium-phosphate homeostasis, vitamin D, through its hormonal form calcitriol, has been found to be a cell-differentiating factor and antiproliferative agent with actions on a variety of tissues. By influencing gene expression, calcitriol affects many physiological processes, besides calcium-phosphate homeostasis, including muscle and keratinocyte differentiation, blood pressure regulation and the immune response. The widespread expression of 1a-hydroxylating enzyme in peripheral tissues supports the view that it exists to boost intracellular concentrations of calcitriol within some target tissues in order to modulate a unique set of genes. This new information has profound implications for chronic kidney disease (CKD) patients with their tendency to vitamin D substrate insufficiency, coupled with their documented loss of the renal 1a-hydroxylase. Administration of vitamin D receptor activators such as calcitriol have been found to favor the survival of CKD patients mainly improving the cardiovascular system. In recent years, analogs of calcitriol such as paricalcitol, able to activate the vitamin D receptor, have shown additional biologic effects and advantages compared with calcitriol in the treatment of patients with chronic renal failure.


A bedside measure of body composition in Duchenne muscular dystrophy.
In clinical practice, monitoring body composition is a critical component of nutritional assessment and weight management in boys with Duchenne muscular dystrophy. We aimed to evaluate the accuracy of a simple bedside measurement tool for body composition, namely bioelectrical impedance analysis, in boys with Duchenne muscular dystrophy. Measures of fat-free mass were determined using a bioelectrical impedance analysis machine and compared against estimations obtained from a reference body composition model. Additionally, the use of raw impedance values was analyzed using three existing predictive equations for the estimation of fat-free mass. Accuracy of bioelectrical impedance analysis was assessed by comparison against the reference model by calculation of biases and limits of agreement. Body composition was measured in 10 boys with Duchenne muscular dystrophy, mean age 9.01 ± 2.34 years. The bioelectrical impedance analysis machine values of fat-free mass were on average 2.3 ± 14.1 kg higher than reference values. Limits of agreement (based on 95% confidence interval of the mean) were -7.4 to 2.9 kg. There was a significant correlation between the mean fat-free mass and difference in fat-free mass between the bioelectrical impedance analysis machine and the reference model (r = -0.86; P = 0.02) suggesting that the bias was not consistent across the range of measurements. The most accurate predictive equation for the estimation of fat-free mass using raw impedance values was the equation by Pietrobelli et al. (mean difference, -0.7 kg; 95% limits of agreement, -3.5 to 2.0 kg). In a clinical setting, where a rapid assessment of body composition is advantageous, the use of raw impedance values, combined with the equation by Pietrobelli et al., is recommended for the accurate estimation of fat-free mass, in boys with Duchenne muscular dystrophy.


Advancements in Myocardial Infarction Management: Exploring Novel Approaches and Strategies.
In the landscape of healthcare, the management of myocardial infarction (MI) stands as a pivotal challenge and a critical juncture where advancements are reshaping the trajectory of patient care. Myocardial infarction, commonly known as a heart attack, remains a foremost contributor to global morbidity and mortality. Conventional management strategies have historically focused on rapid restoration of blood flow through revascularization techniques. However, the last decade has witnessed a profound transformation, with a burgeoning emphasis on precision medicine and innovative interventions. This contextual backdrop sets the stage for a deep dive into the realm of novel diagnostic modalities, spanning high-sensitivity biomarkers, advanced imaging techniques, and data-driven algorithms. These innovations facilitate not only early detection but also the stratification of patients, paving the way for individualized treatment plans. By targeting the underlying mechanisms of myocardial damage, these interventions hold the promise of attenuating the impact of MI and promoting cardiac regeneration. It examines the integration of telemedicine, wearable devices, and remote monitoring platforms, bridging the gap between patients and caregivers while enabling timely interventions. Additionally, the psychosocial aspects of MI recovery are explored, highlighting the integration of psychological support and lifestyle interventions to enhance long-term well-being. By exploring novel diagnostics, innovative therapies, and holistic patient-centered strategies, it underscores the collaborative efforts of medical practitioners, researchers, and technological pioneers in reshaping the trajectory of MI care. As we stand at the intersection of medical advancement and compassionate patient management, embracing these novel approaches promises a future where the impact of myocardial infarction can be mitigated, and lives can be extended and enriched.


Aftermath of Chlamydia Trachomatis - The Tip of an Iceberg in Female Reproductive Health.
The magnitude of infertile couples worldwide was found to be 60-80 million. Genital infection due to Chlamydia trachomatis (C. trachomatis) is one of the most prevalent sexually transmitted infections (STIs) which may present as PID, leading to ectopic pregnancy, infertility or other adverse health outcomes. This study was done to assess the prevalence of C. trachomatis infections among female patients with infertility using real time PCR (RT-PCR) and to compare the findings of molecular testing with hysterosalpingography (HSG) and ultrasonography (USG). 50 endocervical swabs were collected from women of reproductive age group attending infertility clinic and stored at -80 ºC. DNA extraction was done with Helini bacterial mini spin kit and tested for C. trachomatis DNA by RT-PCR kit. Of the 50 patients, 43 (86%) had primary infertility, and 7 (14%) had secondary infertility. Three (6%) were positive for C. trachomatis by RT-PCR. Two had primary infertility and one had secondary infertility. Routine screening of C. trachomatis even in high-risk populations is not available in developing countries like India. The World Health Organization recommends syndromic approach for case management. Hence, a cost-effective, highly sensitive and specific test is the pressing priority in resource poor settings.


Evidence for fibril-like structure in bovine casein micelles.
The addition of Congo red (CR) dye to diluted raw skim milk resulted in a red shift indicative of the presence of fibril-like structures. Thioflavin T (ThT) is another dye that very specifically binds to protein fibrils, and when added to undiluted raw skim milk, the classic 485 nm fluorescence peak of a ThT-fibril complex was observed. Repeating these experiments with various raw milk components showed that the CR red shift and ThT fluorescence peak were due to the presence of casein micelles, and to a lesser extent, sodium caseinate. Fluorescent peaks were also observed when ThT was added to solutions of purified alpha(S)- and kappa-casein, but not beta-casein, in 0.5 M HEPES buffer (pH = 6.8). The addition of 25 mM Ca(2+) had no effect on beta-casein fluorescence, and significantly reduced the kappacasein peak. However, adding 25 mM Ca(2+) to alpha(S)-casein produced a turbid solution and a 6-fold increase in fluorescence, indicating that the aggregates formed contain fibril-like structure. Casein micelle images obtained by transmission electron microscopy showed the presence of short (7 to 10 nm) fibers cross-linked by dense aggregate junction zones. The observed fibers closely resemble protofibrils, intermediate structures that are observed during the formation of amyloid fibrils.


An Exploratory Investigation of the Palatoglossus Muscle in Children Using Magnetic Resonance Imaging.
There is currently little evidence reporting the typical morphology of the palatoglossus (PG) muscle. The primary purpose of this exploratory study is to determine whether magnetic resonance imaging (MRI) methods used to quantify the morphology of the levator veli palatini (LVP) muscle can be applied to the PG. The secondary purpose is to provide preliminary data regarding the relationship between the LVP and PG muscles in children. Ten children between ages of 4 and 7 years participated in this study. Each participant was scanned using a nonsedated, child-friendly protocol with a T2-weighted, three-dimensional anatomical scan to obtain images of the oropharyngeal anatomy. Custom, oblique-coronal image planes were created to visualize and measure the LVP and PG muscles in their entirety from origin to insertion. Thermo Scientific Amira Software was used to obtain 2D measurements of PG muscle length, width, velar insertion distance, lingual insertion distance, and several angle measurements. The PG ranged from 17.95 to 26.96 mm in length across participants. Velar insertion distance ranged from 17.22 to 30.95 mm. Lingual insertion distance ranged from 26.91 to 36.02 mm. Width ranged from 2.32 to 3.08 mm. The angle formed by the PG and LVP muscle planes ranged from 7.3° to 52.7°. The LVP insertion angle ranged from 42.5° to 75.9°. The PG insertion angle ranged from 16.9° to 52.3°. MRI was successful in visualizing the PG muscle. The PG was consistent in size and shape within an individual participant but varied across the participant cohort.


Disagreement, consensus, and moral integrity.
The Advisory Committee on Human Radiation Experiments experienced some disagreements among its members in the course of its work. An epistemological controversy over the nature and degree of evidence required to draw ethical conclusions pervaded the Committee's deliberations. Other disagreements involved the proper role of a governmental advisory committee and the question of when it is appropriate to notify people that they were unknowing subjects of radiation experiments. In the end, the Committee was able to reach consensus on almost all of its findings and recommendations through a process that preserved the integrity of its members.


Control of Product Distribution and Mechanism by Ligation and Electric Field in the Thermal Activation of Methane.
An unexpected mechanistic switch as well as a change of the product distribution in the thermal gas-phase activation of methane have been identified when diatomic [ZnO].+ is ligated with acetonitrile. Theoretical studies suggest that a strong metal-carbon attraction in the pristine [ZnO].+ species plays an important role in the rebound of the incipient CH3. radical to the metal center, thus permitting the competitive generation of CH3. , OH. , and CH3 OH. This interaction is drastically weakened by a single CH3 CN ligand. As a result, upon ligation the proton-coupled single electron transfer that prevails for [ZnO].+ /CH4 switches to the classical hydrogen-atom-transfer process, thus giving rise to the exclusive expulsion of CH3. . This ligand effect can be modeled quite well by an oriented external electric field of a negative point charge.


Effect of glycyrrhizin on cortisol metabolism in humans.
Urinary cortisol and cortisone excretion were measured after glycyrrhizin administration (225 mg/day, per os) for seven consecutive days in five healthy volunteers. The ratio of plasma cortisol and cortisone during hydrocortisone (0.6 mg/kg i.v.) infusion was also examined. After glycyrrhizin administration, the urinary cortisone to cortisol ratio was significantly decreased, but the ratio of plasma cortisol to cortisone was not significantly altered. Glycyrrhizin inhibited 11 beta-hydroxysteroid dehydrogenase in the kidney and so inhibited the conversion of cortisol to cortisone. The increased cortisol in the kidney could be the cause of the apparent mineralocorticoid excess syndrome induced by glycyrrhizin.


ELAV proteins along evolution: back to the nucleus?
The complex interplay of post-transcriptional regulatory mechanisms mediated by RNA-binding proteins (RBP) at different steps of RNA metabolism is pivotal for the development of the nervous system and the maintenance of adult brain activities. In this review, we will focus on the highly conserved ELAV gene family encoding for neuronal-specific RBPs which are necessary for proper neuronal differentiation and important for synaptic plasticity process. In the evolution from Drosophila to man, ELAV proteins seem to have changed their biological functions in relation to their different subcellular localization. While in Drosophila, they are localized in the nuclear compartment of neuronal cells and regulate splicing and polyadenylation, in mammals, the neuronal ELAV proteins are mainly present in the cytoplasm where they participate in regulating mRNA target stability, translation and transport into neurites. However, recent data indicate that the mammalian ELAV RBPs also have nuclear activities, similarly to their fly counterpart, being them able to continuously shuttle between the cytoplasm and the nucleus. Here, we will review and comment on all the biological functions associated with neuronal ELAV proteins along evolution and will show that the post-transcriptional regulatory network mediated by these RBPs in the brain is highly complex and only at an initial stage of being fully understood. This article is part of a Special Issue entitled 'RNA and splicing regulation in neurodegeneration'.


Sphagnum Moss as an Indicator of Contemporary Rates of Atmospheric Dust Deposition in the Athabasca Bituminous Sands Region.
Sphagnum moss was collected from ombrotrophic (rain-fed) peat bogs to quantify dust emissions from the open-pit mining and upgrading of Athabasca bituminous sands (ABS). A total of 30 bogs were sampled in the ABS region, and 5 were sampled in central Alberta. Ash was separated into the acid-insoluble ash (AIA) and acid-soluble ash (ASA) fractions using HCl. The AIA concentrations increase toward industry from 0.4 ± 0.5% to 4.7 ± 2.0% over a distance of 30 km; the control site at the Utikuma Region Study Area (URSA) yielded 0.29 ± 0.07% (n = 30). Mass accumulations rates showed similar spatial variation. The morphology and mineralogy of the AIA particles were studied using scanning electron microscopy and energy-dispersive X-ray analysis and the particle size distributions using optical methods. Particle size was more variable in moss closer to industry. Major ions in the ASA fraction showed elevated accumulation rates of Ca, K, Fe, Mg, P, and S, with P being up to 5 times greater in samples nearest industry compared to those in distal locations. Given that P has been regarded as the growth-limiting nutrient in bogs, fertilization of nutrient-poor ecosystems, such as these from fugitive emissions of dusts from open-pit mining, may have long-term ecological ramifications.


Unfulfilled expectations to services offered at primary health care facilities: experiences of caretakers of underfive children in rural Tanzania.
There is growing evidence that patients frequently bypass primary health care (PHC) facilities in favour of higher level hospitals regardless of substantial additional time and costs. Among the reasons given for bypassing are poor services (including lack of drugs and diagnostic facilities) and lack of trust in health workers. The World Health Report 2008 "PHC now more than ever" pointed to the importance of organizing health services around people's needs and expectations as one of the four main issues of PHC reforms. There is limited documentation of user's expectations to services offered at PHC facilities. The current study is a community extension of a hospital-based survey that showed a high bypassing frequency of PHC facilities among caretakers seeking care for their underfive children at two district hospitals. We aimed to explore caretakers' perceptions and expectations to services offered at PHC facilities in their area with reference to their experiences seeking care at such facilities. We conducted four community-based focus group discussions (FGD's) with 47 caretakers of underfive children in Muheza district of Tanga region, Tanzania in October 2009. Lack of clinical examinations and laboratory tests, combined with shortage of drugs and health workers, were common experiences. Across all the focus group discussions, unpleasant health workers' behaviors, lack of urgency and unnecessary delays were major complaints. In some places, unauthorized fees reduced access to services. The study revealed significant disappointments among caretakers with regard to the quality of services offered at PHC facilities in their areas, with implications for their utilization and proper functioning of the referral system. Practices regarding partial drugs administrations, skipping of injections, unofficial payments and consultations by unskilled health care providers need urgent action. There is also a need for proper accountability mechanisms to govern appropriate allocation and monitoring of health care resources and services in Tanzania.


Congo Red affects the growth, morphology and activity of glucosamine-6-phosphate synthase in the human pathogenic fungus Sporothrix schenckii.
Sporothrix schenckii is the etiological agent of sporotrichosis, a mycosis of humans and other mammals. Little is known about the responses of this thermodimorphic pathogen to perturbations in the cell wall (CW) by different stress conditions. Here we describe the effect of Congo Red (CR) on the fungal growth, morphogenesis and activity of glucosamine-6-phosphate (GlcN-6-P) synthase. Under conditions of yeast development, 15 µM CR abolished conidia (CN) germination, but when yeast cells were first obtained in the absence of the dye and then post-incubated in its presence, yeasts rapidly differentiated into mycelial cells. On the other hand, under conditions of mycelium development, 150 µM CR did not affect CN germination, but filamentous cells underwent structural changes characterized by a distorted CW contour, the loss of polarity and the formation of red-pigmented, hyphal globose structures. Under these conditions, CR also induced a significant and transient increase in the activity of GlcN-6-P synthase, an essential enzyme in CW biogenesis.


A Simple Model of Protein Domain Swapping in Crowded Cellular Environments.
Domain swapping in proteins is an important mechanism of functional and structural innovation. However, despite its ubiquity and importance, the physical mechanisms that lead to domain swapping are poorly understood. Here, we present a simple two-dimensional coarse-grained model of protein domain swapping in the cytoplasm. In our model, two-domain proteins partially unfold and diffuse in continuous space. Monte Carlo multiprotein simulations of the model reveal that domain swapping occurs at intermediate temperatures, whereas folded dimers and folded monomers prevail at low temperatures, and partially unfolded monomers predominate at high temperatures. We use a simplified amino acid alphabet consisting of four residue types, and find that the oligomeric state at a given temperature depends on the sequence of the protein. We also show that hinge strain between domains can promote domain swapping, consistent with experimental observations for real proteins. Domain swapping depends nonmonotonically on the protein concentration, with domain-swapped dimers occurring at intermediate concentrations and nonspecific interactions between partially unfolded proteins occurring at high concentrations. For folded proteins, we recover the result obtained in three-dimensional lattice simulations, i.e., that functional dimerization is most prevalent at intermediate temperatures and nonspecific interactions increase at low temperatures.


Predictors of pain, urinary symptoms and quality of life in patients with chronic pelvic pain syndrome (CPPS): A prospective 12-month follow-up study.
Chronic pelvic pain syndrome (CPPS) can affect both men and women and often causes substantial impairment to quality of life. Although cross-sectional studies have suggested that psychosocial aspects may constitute important factors in the etiology and maintenance of CPPS, longitudinal studies are rare. Therefore, the present study examines psychosocial factors as prospective predictors of pain intensity, urinary symptoms and impediments to quality of life in men and women with CPPS. Data were collected from patients during visits to a specialized, interdisciplinary outpatient clinic and after 12 months. Outcomes included pain intensity, urinary symptoms and impediments to quality of life, all of which were measured with the NIH-CPSI. Age, sex, depressive-anxious symptomatology (PHQ-ADS), pain catastrophizing (PCS), health anxiety (WI-7) and social support (FSozU) were examined as predictors in multivariate linear regressions. Data from 109 patients (59.6% female; age M = 49.3, SD = 16.7) were analyzed. Pain severity (β = .30, p = .004), age (β = .22, p = .02), urinary symptoms (β = .24, p = .01) and depressive-anxious symptomatology (β = .29, p = .009) at baseline emerged as predictors of pain at follow-up. Urinary symptoms were predicted by urinary symptoms (β = .53, p < .001) and depressive-anxious symptomatology (β = .25, p = .01) at baseline; impediments to quality of life were predicted by depressive-anxious symptomatology (β = .27, p = .01). Psychological factors, especially depressive-anxious symptomatology, predict CPPS-specific symptom severity and impediments to quality of life after 12 months and thus substantially contribute to the chronification of CPPS. It is recommended to address anxiety and depression in patients with CPPS as early as possible in biopsychosocially oriented treatment approaches.


Jun NH2-terminal kinase is constitutively activated in T cells transformed by the intracellular parasite Theileria parva.
When T cells become infected by the parasite Theileria parva, they acquire a transformed phenotype and no longer require antigen-specific stimulation or exogenous growth factors. This is accompanied by constitutive interleukin 2 (IL-2) and IL-2 receptor expression. Transformation can be reversed entirely by elimination of the parasites using the specific drug BW720c. Extracellular signal-regulated kinase and jun NH2-terminal kinase (JNK) are members of the mitogen-activated protein kinase family, which play a central role in the regulation of cellular differentiation and proliferation and also participate in the regulation of IL-2 and IL-2 receptor gene expression. T. parva was found to induce an unorthodox pattern of mitogen-activated protein kinase expression in infected T cells. JNK-1 and JNK-2 are constitutively active in a parasite-dependent manner, but have altered properties. In contrast, extracellular signal-regulated kinase-2 is not activated even though its activation pathway is functionally intact. Different components of the T cell receptor (TCR)-dependent signal transduction pathways also were examined. The TCRzeta or CD3epsilon chains were found not to be phosphorylated and T. parva-transformed T cells were resistant to inhibitors that block the early steps of T cell activation. Compounds that inhibit the progression of T cells to proliferation, however, were inhibitory. Our data provide the first example, to our knowledge, for parasite-mediated JNK activation, and our findings strongly suggest that T. parva not only lifts the requirement for antigenic stimulation but also entirely bypasses early TCR-dependent signal transduction pathways to induce continuous proliferation.


Inferring Parameters of the Distribution of Fitness Effects of New Mutations When Beneficial Mutations Are Strongly Advantageous and Rare.
Characterizing the distribution of fitness effects (DFE) for new mutations is central in evolutionary genetics. Analysis of molecular data under the McDonald-Kreitman test has suggested that adaptive substitutions make a substantial contribution to between-species divergence. Methods have been proposed to estimate the parameters of the distribution of fitness effects for positively selected mutations from the unfolded site frequency spectrum (uSFS). Such methods perform well when beneficial mutations are mildly selected and frequent. However, when beneficial mutations are strongly selected and rare, they may make little contribution to standing variation and will thus be difficult to detect from the uSFS. In this study, I analyze uSFS data from simulated populations subject to advantageous mutations with effects on fitness ranging from mildly to strongly beneficial. As expected, frequent, mildly beneficial mutations contribute substantially to standing genetic variation and parameters are accurately recovered from the uSFS. However, when advantageous mutations are strongly selected and rare, there are very few segregating in populations at any one time. Fitting the uSFS in such cases leads to underestimates of the strength of positive selection and may lead researchers to false conclusions regarding the relative contribution adaptive mutations make to molecular evolution. Fortunately, the parameters for the distribution of fitness effects for harmful mutations are estimated with high accuracy and precision. The results from this study suggest that the parameters of positively selected mutations obtained by analysis of the uSFS should be treated with caution and that variability at linked sites should be used in conjunction with standing variability to estimate parameters of the distribution of fitness effects in the future.


Harmful dinoflagellate Ostreopsis cf. ovata Fukuyo: detection of ovatoxins in field samples and cell immunolocalization using antipalytoxin antibodies.
Ostreopsis cf. ovata, a benthic dinoflagellate often blooming along the Mediterranean coasts, has been associated with toxic events ranging from dyspnea to mild dermatitis. In late September 2009, an Ostreopsis cf. ovata bloom occurred in the Gulf of Trieste (Northern Adriatic Sea; Italy), causing pruritus and mild dermatitis in beachgoers. An integrated study was initiated to characterize Ostreopsis cells by light and confocal microscopy, PCR techniques, immunocytochemistry, and high resolution liquid chromatography-mass spectrometry (HR LC-MS). The presence of Ostreopsis cf. ovata of the Atlantic/Mediterranean clade was unambiguously established by morphological and genetic analyses in field samples. Several palytoxin-like compounds (ovatoxin-a,-b,-c,-d,-e) were identified by HR LC-MS, ovatoxin-a being the most abundant (45-64 pg/cell). Surprisingly, no palytoxin was detected. For the first time, monoclonal and polyclonal antipalytoxin antibodies revealed the intracellular cytoplasmic localization of ovatoxins, suggesting their cross-reactivity with these antibodies. Since harmful dinoflagellates do not always produce toxins, the immunocytochemical localization of ovatoxins, although qualitative, can provide an early warning for toxic Ostreopsis cells before their massive diffusion and/or concentration in seafood.


Some ethical dimensions of cochlear implantation for deaf children and their families.
A major source of controversy between Deaf people and those who support a "social/cultural" view of Deafness as "a life to be lived" and those who see deafness within a "medical model" as a "condition to be cured" has been over the cochlear implantation of young deaf children. Recent research has shown that there are noticeable inequities in access to such procedures in western countries; inequities that give rise to the need for informed public policy discussions. It has also found that parents of newly diagnosed deaf children are not provided with access to all the possibilities for their children-including that of a "Deaf life." How this information can be provided to parents and the public via widespread discussions in the media and elsewhere and involving Deaf people in the implantation counseling process is an issue that needs to be addressed by those responsible for implantation programs.


High prevalence of mucosa-associated E. coli producing cyclomodulin and genotoxin in colon cancer.
Some Escherichia coli strains produce toxins designated cyclomodulins (CMs) which interfere with the eukaryotic cell cycle of host cells, suggesting a possible link between these bacteria and cancers. There are relatively few data available concerning the colonization of colon tumors by cyclomodulin- and genotoxic-producing E. coli. We did a qualitative and phylogenetic analysis of mucosa-associated E. coli harboring cyclomodulin-encoding genes from 38 patients with colorectal cancer (CRC) and 31 with diverticulosis. The functionality of these genes was investigated on cell cultures and the genotoxic activity of strains devoid of known CM-encoding gene was investigated. Results showed a higher prevalence of B2 phylogroup E. coli harboring the colibatin-producing genes in biopsies of patients with CRC (55.3%) than in those of patients with diverticulosis (19.3%), (p<0.01). Likewise, a higher prevalence of B2 E. coli harboring the CNF1-encoding genes in biopsies of patients with CRC (39.5%) than in those of patients with diverticulosis (12.9%), (p = 0.01). Functional analysis revealed that the majority of these genes were functional. Analysis of the ability of E. coli to adhere to intestinal epithelial cells Int-407 indicated that highly adherent E. coli strains mostly belonged to A and D phylogroups, whatever the origin of the strains (CRC or diverticulosis), and that most E. coli strains belonging to B2 phylogroup displayed very low levels of adhesion. In addition, 27.6% (n = 21/76) E. coli strains devoid of known cyclomodulin-encoding genes induced DNA damage in vitro, as assessed by the comet assay. In contrast to cyclomodulin-producing E. coli, these strains mainly belonged to A or D E. coli phylogroups, and exhibited a non significant difference in the distribution of CRC and diverticulosis specimens (22% versus 32.5%, p = 0.91). In conclusion, cyclomodulin-producing E. coli belonging mostly to B2 phylogroup colonize the colonic mucosa of patients with CRC.


Effects of 4-MA, a potent inhibitor of 5 alpha-reductase, on 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase activity in guinea pig adrenals.
Previous reports have shown that 17 beta-N,N-Diethylcarbamoyl-4-methyl-4-aza- 5 alpha-androstan-3-one (4-MA), a synthetic inhibitor of 5 alpha-reductase, exerts an inhibitory effect on 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta-HSD) activity. To characterize further the effects of 4-MA on steroidogenesis, guinea pig fasciculata-glomerulosa cells in primary culture were treated for 24 h with 4-MA. Our data indicate that 4-MA reduced 3 beta-HSD activity in cultured adrenal cells but had no effect on the activities of 11-hydroxylase, 21-hydroxylase, 17-hydroxylase, and 17,20-lyase. Be decreasing the conversion of pregnenolone into progesterone or 17-hydroxypregnenolone into 17-hydroxyprogesterone, 4-MA caused the steroidogenic pathway to shift toward the production of dehydroepiandrosterone. Despite the presence of 4-MA, androstenedione and 11 beta-hydroxyandrostenedione were produced at levels exceeding the control levels. In the presence of ACTH and 4-MA, cortisol production was inhibited by 90% whereas androstenedione and 11 beta-hydroxyandrostenedione were reduced by only 40%. The effect of the compound was reversed by washing the adrenal cells with medium, thus suggesting a direct action of 4-MA on the enzyme itself. In summary, our data indicate that 4-MA markedly reduces the production of cortisol in the adrenals and partially alters the formation of C-19 steroids. It is important to consider this finding in the use of 4-azasteroids in the treatment of prostate cancer, which was previously found to be sensitive to secretion of adrenal C-19 steroids.


Embryonic fissure and photoreceptor differentiation in the eye of adult Garra rufa Heckel 1843 (Cyprinidae, Teleostei).
The retina of the adult teleost Garra rufa retains a curved, open embryonic fissure indicating an asymmetrical postembryonic retinal growth. Undifferentiated, oval photoreceptors are observed on both sides of the middle of the fissure with their larger diameter running parallel to the fissure to which they may attach by desmosomes. They detach from the fissure, rotate to become perpendicular to it and begin an active process of differentiation as they slide along the temporal side of the outer half of the fissure. This process is divided into stages for simplicity. The photoreceptors develop stumpy inner segments extending into a ventricular space that appears between the retinal pigment epithelium and the photoreceptors. Calycal processes arise from the inner segments and the distal centriole of each photoreceptor forms a connecting cilium. The proximal centriole is retained for some time after the outer segment develops. The formation of rod spherules and cone pedicles takes place almost concomitantly with the outer segments. Double cones appear first as single cones before pairing. One or more of the principal cone mitochondria accumulate electron-dense material and merge to form the ellipsosome. The retinal pigment epithelium undergoes a parallel differentiation. The developmental events described in the present work conform those recorded in embryonic teleostean retinas.