Application of different molecular techniques for characterization of catalase-positive cocci isolated from sucuk.
This study was carried out for the characterization and discrimination of the indigenous Gram positive, catalase-positive cocci (GCC) population in sucuk, a traditional Turkish dry-fermented sausage. Sucuk samples, produced by the traditional method without starter culture were collected from 8 local producers in Kayseri/Turkey and a total of 116 GCC isolates were identified by using different molecular techniques. Two different molecular fingerprinting methods; namely, randomly amplified polymorphic DNA-PCR (RAPD-PCR) and repetitive extragenic palindrome-PCR (rep-PCR), were used for the clustering of isolates and identification at species level was carried out by full length sequencing of 16S rDNA. Combining the results obtained from molecular fingerprinting and 16S rDNA sequencing showed that the dominant GCC species isolated from the sucuk samples was Staphylococcus saprophyticus followed by Staphylococcus succinus and Staphylococcus equorum belonging to the Staphylococcus genus. Real-time PCR DNA melting curve analysis and high-resolution melting (HRM) analysis targeting the V1 + V3 regions of 16S rDNA were also applied for the discrimination of isolates belonging to different species. It was observed statistically different Tm values and species-specific HRM profiles for all except 2 species (S. saprophyticus and Staphylococcus xylosus) that have high 16S rDNA sequence similarity. The combination of rep-PCR and/or PCR-RAPD with 16S rRNA gene sequencing was an efficient approach for the characterization and identification of the GCC population in spontaneously fermented sucuk. On the other hand, intercalating dye assays were found to be a simple and very promising technique for the differentiation of the GCC population at species level.


Kinetic analysis of interaction of BRCA1 tandem breast cancer c-terminal domains with phosphorylated peptides reveals two binding conformations.
Tandem breast cancer C-terminal (BRCT) domains, present in many DNA repair and cell cycle checkpoint signaling proteins, are phosphoprotein binding modules. The best-characterized tandem BRCT domains to date are from the protein BRCA1 (BRCA1-BRCT), an E3 ubiquitin ligase that has been linked to breast and ovarian cancer. While X-ray crystallography and NMR spectroscopy studies have uncovered the structural determinants of specificity of BRCA1-BRCT for phosphorylated peptides, a detailed kinetic and thermodynamic characterization of the interaction is also required to understand how structure and dynamics are connected and therefore better probe the mechanism of phosphopeptide recognition by BRCT domains. Through a global analysis of binding kinetics data obtained from surface plasmon resonance (SPR) and stopped-flow fluorescence spectroscopy, we show that the recognition mechanism is complex and best modeled by two equilibrium conformations of BRCA1-BRCT in the free state that both interact with a phosphopeptide, with dissociation constants ( K d) in the micromolar range. We show that the apparent global dissociation constant derived from this kinetic analysis is similar to the K d values measured using steady-state SPR, isothermal titration calorimetry, and fluorescence anisotropy. The dynamic nature of BRCA1-BRCT may facilitate the binding of BRCA1 to different phosphorylated protein targets.


A behavioral analysis of dentate gyrus function.
Computational models of the dentate gyrus (DG) have suggested based on anatomical, electrophysiological, and computer simulation data that the DG plays an important role in learning and memory by processing and representing spatial information on the basis of conjunctive encoding, pattern separation, and encoding of spatial information in conjunction with the CA3. Behavioral evidence supports a role for the DG in mnemonic processing of spatial information based on the operation of conjunctive encoding of multiple sensory inputs, pattern separation of spatial (especially metric) information, and subsequent encoding in cooperation with CA3. A potential role of the DG in mediating processes, such as recall of sequential information and short-term memory as well as temporal order for remote memory, are also discussed.


Mustard seeds (Sinapis Alba Linn) attenuate azoxymethane-induced colon carcinogenesis.
Mustard seeds (MS), which are consumed in considerable amounts by the Japanese people that, interestingly, have the longest life expectancy in the world, are known to contain a number of yet not fully defined but quite powerful anti-oxidants. A suspension of extracted MS was found to suppress oxidized-LDL-induced macrophage respiratory burst in vitro, to prevent growth, and to induce apoptotic death of SW480 cells (a human colon cancer cell line), while no such effects were found for normal 3T3 cells. A diet enriched with MS decreased plasma levels of the lipid peroxidation product malonaldehyde in mice exposed to the colon cancer-inducer azoxymethane (AOM). Such a diet also dose-dependently enhanced the activity of several anti-oxidant enzymes, such as superoxide dismutase (SOD), catalase, and GSH-peroxidase and, moreover, reduced AOM-mediated formation of colon adenomas by about 50%. Further studies are required to detail and explore the beneficial effects of MS and their rich content of powerful anti-oxidants.


Extensive NMRD studies of Ni(II) salt solutions in water and water-glycerol mixtures.
Aqueous solutions of simple nickel(II) salts are a classical test case for theories of the paramagnetic relaxation enhancement (PRE) and its dependence on the magnetic field (nuclear magnetic relaxation dispersion, NMRD), going back to late fifties. We present here new experimental data, extending the NMRD range up to 21T (900 MHz). In addition to salt solutions in (acidified) water, we have also measured on solutions containing glycerol. The aqueous solution data do not show any significant changes compared to the earlier experiments. The interpretation, based on the general ("slow-motion") theory is also similar to the earlier work from our laboratory. The NMRD-data in mixed solvents are qualitatively different, indicating that the glycerol not only changes the solution viscosity, but may also enter the first coordination sphere of the metal ion, resulting in lower symmetry complexes, characterized by non-vanishing averaged zero-field splitting. This hypothesis is corroborated by molecular dynamics simulations. A strategy appropriate for interpreting the NMRD-data for the chemically complicated systems of this type is proposed.


The history of urologic pathology: an overview.
This article begins with the testis and a legendary figure, Sir Astley Cooper, who wrote an early text on the organ. The early 20th century saw the first major development, the description of the seminoma by the French investigator Maurice Chevassu, but the pace of knowledge did not accelerate until after World War II with a major article from the Armed Forces Institute of Pathology (AFIP) by Nathan B. Friedman and Robert A. Moore, soon followed by the first series testis fascicle by Frank J. Dixon and Moore. Other noteworthy contributions were made by two masters of gonadal pathology, Gunnar Teilum and Robert E. Scully. In the 1970s, Niels E. Skakkebaek played a seminal role in elaborating in-situ neoplasia of the testis. The school of British testicular tumour authored, in the mid-1970s, under the editorship of Roger C. B. Pugh, one of the best texts on testicular pathology. Advances in more recent years have been largely spearheaded by Thomas M. Ulbright of the Indiana University School of Medicine. Observations on the prostate gland date back to Andreas Vesalius and William Cheselden, the latter appearing to have introduced the word for the gland. Note is made of contributions on the anatomy and histology of the gland by Oswald Lowsley, L. M. Franks, and John McNeal. Diagnosing carcinoma of the prostate was brought into the modern age in a landmark 1953 article by Robert S. Totten et al. In the 1960s, Donald F. Gleason introduced a grading system that is now in use worldwide. The topic of premalignant lesions has been well established only for approximately three decades, based initially on the work of Dr McNeal and David G. Bostwick. One of the first to write a book on the bladder was the remarkable British surgeon-pathologist Sir Henry Thompson. Workers at the AFIP, including Colonel James E. Ash and Fatallah K. Mostofi, wrote many outstanding articles on bladder pathology. The roles of other institutions, such as Johns Hopkins University, the Mayo Clinic, and St Peter's Hospital Institute of Urology, London, and those who worked there are noted. Knowledge of the pathology of the urachus dates largely back to the remarkable book on the topic in 1916 by the Hopkins investigator Thomas S. Cullen. Information on renal tumours dates largely to the work of Paul Grawitz, but awareness of the many variants of renal cell carcinoma in general was slow to evolve, and has only accelerated in recent years. The AFIP group of Dr Mostofi, ably assisted by Colonel Charles J. Davis and Isabell A. Sesterhenn, has contributed to knowledge of renal neoplasia with articles of note on oncocytoma, metanephric adenoma, and medullary carcinoma. In the mid-1980s, the German workers Wolfgang Thoenes and Stephan Störkel recognised the distinctive tumour known as chromophobe renal cell carcinoma. Work on renal tumours in the young owes much to J. Bruce Beckwith. The observational talents of numerous investigators have, in just over a century, advanced our knowledge of diseases of the urinary tract and testis remarkably.


[Solid liver mass. Studies with nuclear medicine].
We present the case of a 23 year old female with two incidentally detected hepatic mass that have not clinically o radiologically specific findings. Nuclear medicine tracers, including colloids and hepatobiliary agents showing the characteristic findings of focal nodular hyperplasia: Hypervascularization, normal uptake of colloids, accumulation of hepatobiliary tracer and hot spots due to the retention of this tracer during the clearance fhase. The patient was underwent hepatectomy. The examination of surgical specimen revealed focal nodular hyperplasia. The scintigraphic studies could be an useful tool in the noninvasive diagnosis of liver masses.


[Distant metastases on acinic cell carcinoma of the parotid gland after 12 years symptom-free interval].
Acinic cell carcinoma of parotid gland as cause of distant metastases are rare. The patient was a 60-year-old woman who had in 1993 a acinic cell carcinoma of right parotid gland. Tumour can be resected through total parotidectomy with facial nerve anastomosis and modified radical neck dissection (T (3) N (2b) M (0)). Since the operation the patient has remained symptom-free without any sign of tumour recurrence. After 12 years the patient noted swelling in the region of sternum and biopsy was necessary. Histologically and immunohistochemically the diagnosis of distant metastase on acinic carcinoma of the parotid gland was confirmed.


Effective deployment of technology-supported management of chronic respiratory conditions: a call for stakeholder engagement.
Healthcare systems are under increasing strain, predominantly due to chronic non-communicable diseases. Connected healthcare technologies are becoming ever more capable and their components cheaper. These innovations could facilitate both self-management and more efficient use of healthcare resources for common respiratory diseases such as asthma and chronic obstructive pulmonary disease. However, newer technologies can only facilitate major changes in practice, and cannot accomplish them in isolation. There are now large numbers of devices and software offerings available. However, the potential of such technologies is not being realised due to limited engagement with the public, clinicians and providers, and a relative paucity of evidence describing elements of best practice in this complex and evolving environment. Indeed, there are clear examples of wasted resources and potential harm. We therefore call on interested parties to work collaboratively to begin to realize the potential benefits and reduce the risks of connected technologies through change in practice. We highlight key areas where such partnership can facilitate the effective and safe use of technology in chronic respiratory care: developing data standards and fostering inter-operability, making collaborative testing facilities available at scale for small to medium enterprises, developing and promoting new adaptive trial designs, developing robust health economic models, agreeing expedited approval pathways, and detailed planning of dissemination to use. The increasing capability and availability of connected technologies in respiratory care offers great opportunities and significant risks. A co-ordinated collaborative approach is needed to realize these benefits at scale. Using newer technologies to revolutionize practice relies on widespread engagement and cannot be delivered by a minority of interested specialists. Failure to engage risks a costly and inefficient chapter in respiratory care.


[How can I keep up with my literature?].
To keep up with the literature, the author and his wife have personal home subscriptions to the 'big five' general medical journals, two top general science journals, and close to a dozen specialist journals. Two of the general medical journals are regularly read in more depth. Since computerized search systems permit almost immediate retrieval on whatever subject at whatever work location, the author keeps no filing system of the items of interest. For three (slowly growing) subjects that transgress the boundaries of the usual keywords of search systems, copies of papers are collected in piles: (a) important methodological papers (which may have been published anywhere, on any subject), (b) papers about concepts of the origin and evolution of diseases, and (c) papers of strong general interest. The disadvantage of not keeping track of the literature by a personal filing system is that the author regularly forgets about papers and books he already has; the advantage is that no time is spent keeping a personal filing system.


Vasopressors: Do they have any role in hemorrhagic shock?
The priority in the management of patients with traumatic hemorrhagic shock is to control the bleeding with simultaneous volume resuscitation to maintain adequate tissue perfusion. Fluid replacement remains the mainstay of initial resuscitation in hemorrhagic shock. Traditionally, vasopressors are contraindicated in the early management of hemorrhagic shock due to their deleterious consequences, although vasopressors may have a role in resuscitation when vasoplegic shock ensues and blood pressure cannot be maintained by fluids alone. Use of vasopressors is not recommended according to the Advanced Trauma Life SupportR management principles. The role of vasopressors remains controversial with no clear guidelines on the timing, type, and dose of these drugs in hemorrhagic shock. Among vasopressors, norepinephrine and vasopressin have been used in the majority of the trials, although not many studies compare the effect of these two on long-term survival in trauma patients. This article reviews the pathophysiology of hemorrhagic shock, adverse effects of fluid resuscitation, and the various experimental and clinical studies on the use of vasopressors in the early phase of resuscitation in hemorrhagic shock.


Preoperative heparin therapy causes immune-mediated hypotension upon heparin administration for cardiac surgery.
To evaluate whether patients with positive or negative heparin antibodies who received heparin preoperatively by continuous infusion developed cardiovascular changes upon heparin administration prior to cardiopulmonary bypass. Clinical trial. Single institution, academic hospital. Eighty (80) patients with good ventricular function on low-dose heparin infusion prior to surgery. Patients were divided into 2 equal groups: group A had negative heparin antibodies (% ratio < 0.26), group B had positive heparin antibodies (% ratio > 1.2). All patients received heparin, 400 units/kg, prior to institution of cardiopulmonary bypass. Cardiovascular changes, activated coagulation time (ACT), and histamine levels were measured before and 5 minutes after administration of heparin. Platelets also were counted before and 6 hours after surgery. Significant hypotension and decreased cardiac index occurred in patients with positive heparin antibodies who received heparin prior to cardiac surgery. Histamine levels increased significantly 5 minutes after heparin administration. Significant thrombocytopenia occurred 6 hours after surgery in group B patients. There was a good correlation between heparin antibodies, histamine levels, thrombocytopenia and cardiovascular changes. Group B patients also had heparin resistance as manifested by a lower ACT after the loading doses of heparin. Patients with positive heparin antibodies pretreated with heparin prior to surgery developed a type of immune-mediated cardiovascular changes and postoperative thrombocytopenia.


Selective reduction of post-selection CD8 thymocyte proliferation in IL-15Rα deficient mice.
Peripheral CD8(+) T cells are defective in both IL-15 and IL-15Rα knock-out (KO) mice; however, whether IL-15/IL-15Rα deficiency has a similar effect on CD8 single-positive (SP) thymocytes remains unclear. In this study, we investigated whether the absence of IL-15 transpresentation in IL-15Rα KO mice results in a defect in thymic CD8 single positive (SP) TCR(hi) thymocytes. Comparison of CD8SP TCR(hi) thymocytes from IL-15Rα KO mice with their wild type (WT) counterparts by flow cytometry showed a significant reduction in the percentage of CD69(-) CD8SP TCR(hi) thymocytes, which represent thymic premigrants. In addition, analysis of in vivo 5-bromo-2-deoxyuridine (BrdU) incorporation demonstrated that premigrant expansion of CD8SP TCR(hi) thymocytes was reduced in IL-15Rα KO mice. The presence of IL-15 transpresentation-dependent expansion in CD8SP TCR(hi) thymocytes was assessed by culturing total thymocytes in IL-15Rα-Fc fusion protein-pre-bound plates that were pre-incubated with IL-15 to mimic IL-15 transpresentation in vitro. The results demonstrated that CD8SP thymocytes selectively outgrew other thymic subsets. The contribution of the newly divided CD8SP thymocytes to the peripheral CD8(+) T cell pool was examined using double labeling with intrathymically injected FITC and intravenously injected BrdU. A marked decrease in FITC(+) BrdU(+) CD8(+) T cells was observed in the IL-15Rα KO lymph nodes. Through these experiments, we identified an IL-15 transpresentation-dependent proliferation process selective for the mature CD8SP premigrant subpopulation. Importantly, this process may contribute to the maintenance of the normal peripheral CD8(+) T cell pool.


Transforming growth factor-beta(1), -beta(2), -beta(3) and their type I and II receptors in human term placenta.
Expression of transforming growth factor (TGF)-beta(3) as well as cellular localization of TGF-beta receptors has not been demonstrated in placenta. TGF-beta receptor type I (RI) and type II (RII) are required to transmit TGF-beta signals, therefore the determination of cells expressing both receptors in concert is necessary to identify target cells for TGF-beta. We investigated presence and localization of TGF-betas and their receptors (RI, RII) in human term placenta using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. RT-PCR showed that messenger RNA for TGF-beta(1), -beta(2), -beta(3), and RI and RII is present in human term placenta. We found intense staining for all TGF-beta isoform and receptor proteins in the syncytiotrophoblastic layer, chorionic plate, and in cells of the extravillous trophoblasts using immunohistochemistry. The simultaneous expression of ligands and their receptors support the hypothesis that TGF-beta may play an important role in regulating growth, differentiation, and function of the human placenta.


BDNF and the Antidepressant Effects of Ketamine and Propofol in Electroconvulsive Therapy: A Preliminary Study.
Ketamine and propofol have become increasingly popular in electroconvulsive therapy (ECT) anaesthesia. This study was conducted to examine whether changes in serum levels of brain-derived neurotrophic factor (BDNF) are associated with the antidepressant effects of ketofol, a combination of ketamine and propofol, in ECT for patients with treatment-resistant depression (TRD). Thirty patients with TRD (18-65 years) were enrolled and underwent eight ECT sessions with ketamine (0.5 mg/kg) plus propofol (0.5 mg/kg) (ketofol). Symptom severity was monitored using the 17-item Hamilton Depression Rating Scale (HAMD-17) and the Brief Psychiatric Rating Scale (BPRS), and serum levels of BDNF were examined by enzyme-linked immunosorbent assay (ELISA) at baseline and after 2, 4, and 8 ECT treatments. Serum levels of BDNF were also collected from thirty healthy controls. At baseline, there were no significant differences in serum levels of BDNF between patients with TRD and healthy controls. The response and remission rates in patients with TRD were 100% (30/30) and 53.3% (16/30) after ECT treatment, respectively. Despite a significant reduction in HAMD-17 and BPRS scores after ECT, no changes in serum levels of BDNF were observed after ECT treatment when compared to baseline. No association was found between serum levels of BDNF and changes in illness severity. Serum levels of BDNF did not represent a suitable candidate biomarker for determining the antidepressant effects of ketofol during ECT for patients with TRD.


Thermostability of mammalian brain ribosomes and the effects of nucleoside triphosphates on their heat-sensitivity.
Mammalian brain ribosomes were found to be heat-labile. On preincubation of the ribosomes at 37 degrees C, their ability to participate in polypeptide-synthesis reactions was substantially diminished. Despite the sensitivity of ribosomal protein synthesis to heat-inactivation, preincubation resulted in no significant alterations in ribosomal sedimentation profiles or changes in the integrity of the ribosomal RNA. The thermolability of brain ribosomes was shown to be associated with their inability to bind both template RNA and aminoacyl-tRNA. Similar experiments with brain ribosomal subunits demonstrated that the small (40S) subunit was more sensitive to heat-inactivation than the large (60S) subunit. The presence of ATP (1mm) protected ribosomes from thermal inactivation, although this protection was shown to be temporary. The protection appeared to be specific to nucleoside triphosphates, since GTP and UTP also stabilized ribosomes to thermal denaturation whereas nucleoside diphosphates (ADP) and nucleoside monophosphates (AMP and cyclic AMP) did not alter ribosomal sensitivity to heat. Although 1mm concentrations of nucleoside triphosphates protected ribosomes from heat-inactivation, the presence of higher concentrations resulted in complete inactivation of ribosomal activity.


Capitalizing on the recession's effect on hospital RN shortages.
The recent economic recession and slow-paced recovery have contributed to dampening out the shortage of hospital-employed registered nurses (RNs), a shortage that has persisted since the mid-1980s. While national unemployment rates remain relatively high and continue to exert economic pressure on RNs to maintain high levels of employment activity, hospital and nurse executives now have an opportunity to make strategic investments in the organizational infrastructure supporting nursing because once the economy rebounds, RN shortages could easily resurface.


Palaeoclimate change drove diversification among isolated mountain refugia in the Australian arid zone.
Refugia featured prominently in shaping evolutionary trajectories during repeated cycles of glaciation in the Quaternary, particularly in the Northern Hemisphere. The Southern Hemisphere instead experienced cycles of severe aridification but little is known about the temporal presence and role of refugia for arid-adapted biota. Isolated mountain ranges located in the Australian arid zone likely provided refugia for many species following Mio/Pliocene (<15 Ma) aridification; however, the evolutionary consequences of the recent development of widespread sand deserts is largely unknown. To test alternative hypotheses of ancient vs. recent isolation, we generated a 10 gene data set to assess divergence history among saxicolous geckos in the genus Heteronotia that have distributions confined to major rocky ranges in the arid zone. Phylogenetic analyses show that each rocky range harbours a divergent lineage, and substantial intraspecific diversity is likely due to topographic complexity in these areas. Old divergences (~4 Ma) among lineages pre-date the formation of the geologically young sand deserts (<1 Ma), suggesting that Pliocene climate shifts fractured the distributions of biota long before the spread of the deserts.


Sustained delivery of anti-VEGFs from thermogel depots inhibits angiogenesis without the need for multiple injections.
Anti-vascular endothelial growth factor (anti-VEGF) proteins are the gold-standard treatment for posterior eye segment proliferative vascular diseases such as Age-Related Macular Degeneration (AMD) and Diabetic Retinopathy (DR). However, the standard of care requires inconvenient monthly intravitreal injections. This underlies an unmet clinical need to develop alternative solutions for sustained delivery of biologics. In this paper, we demonstrated that anti-VEGFs can be encapsulated by a simple mild process into our polyurethane thermogel depots. By changing the hydrophilic-hydrophobic balance in the copolymer, anti-VEGF release rates can be modulated. The antibody in the thermogel partitions into protein domains which vary in size corresponding to the hydrophilicity balance of the polymer. Anti-VEGFs can be released in a relatively linear manner from the thermogel for up to 40 days in vitro. The encapsulated anti-VEGFs demonstrate anti-angiogenic bioactivity by inhibiting vessel outgrowth in rat ex vivo choroidal explants, and reducing vascular leakage in a VEGF-driven neovascularization rabbit model. In conclusion, we show that these thermogels can be tuned in terms of hydrophilicity and used for sustained delivery of bioactive anti-VEGFs. Physically cross-linked polyurethane thermoresponsive hydrogels could be a promising platform for sustained delivery of biologically active therapeutic proteins.


Textractor: a hybrid system for medications and reason for their prescription extraction from clinical text documents.
OBJECTIVE To describe a new medication information extraction system-Textractor-developed for the 'i2b2 medication extraction challenge'. The development, functionalities, and official evaluation of the system are detailed. Textractor is based on the Apache Unstructured Information Management Architecture (UMIA) framework, and uses methods that are a hybrid between machine learning and pattern matching. Two modules in the system are based on machine learning algorithms, while other modules use regular expressions, rules, and dictionaries, and one module embeds MetaMap Transfer. The official evaluation was based on a reference standard of 251 discharge summaries annotated by all teams participating in the challenge. The metrics used were recall, precision, and the F(1)-measure. They were calculated with exact and inexact matches, and were averaged at the level of systems and documents. The reference metric for this challenge, the system-level overall F(1)-measure, reached about 77% for exact matches, with a recall of 72% and a precision of 83%. Performance was the best with route information (F(1)-measure about 86%), and was good for dosage and frequency information, with F(1)-measures of about 82-85%. Results were not as good for durations, with F(1)-measures of 36-39%, and for reasons, with F(1)-measures of 24-27%. The official evaluation of Textractor for the i2b2 medication extraction challenge demonstrated satisfactory performance. This system was among the 10 best performing systems in this challenge.


Synthesis of Responsive Two-Dimensional Polymers via Self-Assembled DNA Networks.
Despite a growing interest in two-dimensional polymers, their rational synthesis remains a challenge. The solution-phase synthesis of a two-dimensional polymer is reported. A DNA-based monomer self-assembles into a supramolecular network, which is further converted into the covalently linked two-dimensional polymer by anthracene dimerization. The polymers appear as uniform monolayers, as shown by AFM and TEM imaging. Furthermore, they exhibit a pronounced solvent responsivity. The results demonstrate the value of DNA-controlled self-assembly for the formation of two-dimensional polymers in solution.


Enteral-tube-feeding diarrhoea: manipulating the colonic microbiota with probiotics and prebiotics.
Diarrhoea is a common and serious complication of enteral tube feeding. Its pathogenesis involves antibiotic prescription, enteropathogenic colonization and abnormal colonic responses, all of which involve an interaction with the colonic microbiota. Alterations in the colonic microbiota have been identified in patients receiving enteral tube feeding and these changes may be associated with the incidence of diarrhoea. Preventing negative alterations in the colonic microbiota has therefore been investigated as a method of reducing the incidence of diarrhoea. Probiotics and prebiotics may be effective because of their suppression of enteropathogenic colonization, stimulation of immune function and modulation of colonic metabolism. Randomized controlled trials of probiotics have produced contrasting results, although Saccharomyces boulardii has been shown to reduce the incidence of diarrhoea in patients in the intensive care unit receiving enteral tube feeding. Prebiotic fructo-oligosaccharides have been shown to increase the concentration of faecal bifidobacteria in healthy subjects consuming enteral formula, although this finding has not yet been confirmed in patients receiving enteral tube feeding. Furthermore, there are no clinical trials investigating the effect of a prebiotic alone on the incidence of diarrhoea. Further trials of the efficacy of probiotics and prebiotics, alone and in combination, in preventing diarrhoea in this patient group are warranted.


Differences in genomic DNA sequences between pathogenic and nonpathogenic isolates of Entamoeba histolytica identified by polymerase chain reaction.
A lambda gt11 cDNA library was constructed from the poly(A)+ RNA of trophozoites of Entamoeba histolytica HM-1:IMSS strain. The library was immunologically screened with monoclonal antibody 4G6, which is specific for the 30,000-Mr antigen of pathogenic isolates. A 0.7-kb clone was isolated, and its nucleotide sequence was determined. To examine whether this gene was specific for pathogenic isolates, a polymerase chain reaction was performed by using four sets of primers and the genomic DNA of pathogenic and nonpathogenic isolates as templates. Amplified DNAs were detected not only in pathogenic isolates but also in nonpathogenic isolates. However, when sequences of amplified DNA of these isolates were compared, minor differences were observed. By considering the presence or absence of recognition sites of some endonucleases, it was possible to distinguish between the pathogenic and nonpathogenic isolates. When various isolates with different zymodemes were examined by polymerase chain reaction and enzyme digestion, the results of typing were entirely in accord with those of zymodeme analysis. These results indicate that there is dimorphism in the genomic DNA coding the 30,000-Mr antigen of E. histolytica and that the combined use of the polymerase chain reaction and enzyme digestion is a useful strategy for identification of species and determination of pathogenicity.


Response of Antarctic cryoconite microbial communities to light.
Microbial communities on polar glacier surfaces are found dispersed on the ice surface, or concentrated in cryoconite holes and cryolakes, which are accumulations of debris covered by a layer of ice for some or all of the year. The ice lid limits the penetration of photosynthetically available radiation (PAR) to the sediment layer, since the ice attenuates up to 99% of incoming radiation. This suite of field and laboratory experiments demonstrates that PAR is an important control on primary production in cryoconite and cryolake ecosystems. Increased light intensity increased efficiency of primary production in controlled laboratory incubations of debris from the surface of Joyce Glacier, McMurdo Dry Valleys, Antarctica. However, when light intensity was increased to levels near that received on the ice surface, without the protection of an ice lid, efficiency decreased and measurements of photophysiology showed that the communities suffered light stress. The communities are therefore well adapted to low light levels. Comparison with Arctic cryoconite communities, which are typically not covered by an ice lid for the majority of the ablation season, showed that these organisms were also stressed by high light, so they must employ strategies to protect against photodamage.


[Wernicke encephalopathy and Korsakoff's psychosis: clinical-pathophysiological correlation, diagnostics and treatment].
Wernicke's encephalopathy and Korsakoff's psychosis are severe unfavorable forms of alcoholic brain damage with poor prognosis. Thiamine deficiency represents a common cause of both diseases. In many cases, Korsakoff's psychosis develops in the outcome of Wernicke's encephalopathy, which, along with the general etiology, lets talk about a single disease - Wernicke-Korsakoff syndrome, acute (usually reversible) stage of which is Wernicke's encephalopathy and a chronic one (often irreversible) is Korsakoff psychosis. The dramatic paradox of Wernicke's encephalopathy is that in most cases it is difficult to detect, but early diagnosed cases are quite easy to cure. Unrecognized and therefore go untreated Wernicke's encephalopathy is a serious threat to the health and lives of patients, worsens the processes of brain aging and increases the risk of Alzheimer's disease in later life. The basic approach to the treatment of Wernicke-Korsakoff syndrome is long-term parenteral administration of thiamine, often in high doses. As an adjuvant means of therapy memantine is considered.


Injuries to children related to shopping carts.
To describe the epidemiology of shopping cart-related injuries among children and to consider targeted prevention strategies based on these epidemiologic findings. A consecutive series of patients. The emergency department of a large, academic children's hospital. Sixty-two children treated for shopping cart-related injuries during a 15-month period. Children ranged in age from 4 months to 10 years (mean, 2.8 years). Thirty-three children (53%) were boys. Twelve patients (19%) arrived via ambulance. Forty-nine children (79%) had injuries to the head, including one child admitted to the hospital. Eleven children (18%) had fractures, including 5 (8%) with skull fractures. Nine patients (14%) had lacerations, and 30 patients (48%) had superficial injuries (ecchymoses or abrasions). The most common mechanism of injury was falling out of the carts (58% of children), followed by cart tip-overs (26% of children). Injuries caused by falls from the carts occurred across the entire age range, whereas injuries caused by cart tip-overs were most frequent among children 1 year of age or younger. The sitting position was associated with tip-over injuries, and standing in the cart basket was associated with falling from the cart. Shopping cart-related injuries cause serious pediatric morbidity, especially among children younger than 5 years of age, and are potentially fatal. Based on identified age-specific mechanisms of injury, currently used prevention strategies are not sufficient. The use of infant seats and restraining belts is an inadequate strategy for prevention of shopping cart-related injuries among children 1 year of age or younger, because cart tip-over is an important mechanism of injury in this age group. Shopping carts should be redesigned to decrease the tip-over hazard. Transportation of children in shopping carts of current design should be prohibited.


Different effects of thymidine and 5-fluorouracil 2'-deoxyriboside on biosynthetic events in cultured P815Y mast cells.
1. Addition of 2mm-thymidine, although resulting in the cessation of cell division, allows the continuation of phospholipid and protein synthesis and results in an increase in mean cell volume for at least 15h. 2. 5-Fluorouracil 2'-deoxyriboside inhibits cell division but differs from thymidine by inhibiting the synthesis of phospholipid and protein in a more marked manner. 3. The relation between these results and the P815Y cell cycle is discussed.


A meta-analysis on alcohol drinking and gastric cancer risk.
Whether an association between alcohol drinking and gastric cancer risk exists is an open question. In order to provide a definite quantification of the association between alcohol drinking and gastric cancer risk, we conducted a meta-analysis of available data. We carried out a PubMed search of articles published up to June 2010 and identified 44 case-control and 15 cohort studies, including a total of 34 557 gastric cancer cases. We derived meta-analytic estimates using random-effects models, taking into account correlation between estimates. We carried out a dose-risk analysis using nonlinear random-effects meta-regression models. Compared with nondrinkers, the pooled relative risk (RR) was 1.07 [95% confidence interval (CI) 1.01-1.13] for alcohol drinkers and 1.20 (95% CI 1.01-1.44) for heavy alcohol drinkers (≥4 drinks per day). The pooled estimates were apparently higher for gastric noncardia (RR for heavy drinkers=1.17, 95% CI 0.78-1.75) than for gastric cardia (RR=0.99, 95% CI 0.67-1.47) adenocarcinoma. The dose-risk model estimated a RR of 0.95 (95% CI 0.91-0.99) for 10 g/day and 1.14 (95% CI 1.08-1.21) for 50 g/day. This meta-analysis provides definite evidence of a lack of association between moderate alcohol drinking and gastric cancer risk. There was, however, a positive association with heavy alcohol drinking.


Cathelicidin Antimicrobial Peptides with Reduced Activation of Toll-Like Receptor Signaling Have Potent Bactericidal Activity against Colistin-Resistant Bacteria.
The world is at the precipice of a postantibiotic era in which medical procedures and minor injuries can result in bacterial infections that are no longer effectively treated by antibiotics. Cathelicidins are peptides produced by animals to combat bacterial infections and to regulate innate immune responses. However, cathelicidins are potent activators of the inflammatory response. Cathelicidins with reduced proinflammatory activity and potent bactericidal activity in the low micromolar range against Gram-negative bacteria have been identified. Motifs in cathelicidins that impact bactericidal activity and cytotoxicity to human cells have been elucidated and used to generate peptides that have reduced activation of proinflammatory cytokine production and reduced cytotoxicity to human cells. The resultant peptides have bactericidal activities comparable to that of colistin and can kill colistin-resistant bacteria. Cathelicidins are antimicrobial peptides that can also increase inflammatory responses. This combination of activities can cause complications in the treatment of bacterial infections despite the pressing need for new antimicrobials. We have identified cathelicidins with decreased activation of inflammatory responses. The peptides kill Gram-negative bacteria at low micromolar concentrations by binding to and perturbing the integrity of the bacterial membrane. The peptides were also engineered to further decrease lysis of human red blood cells. The peptides have activities comparable to those of the polymyxins, a class of antibiotics to which plasmid-borne resistance is rapidly spreading and can kill colistin-resistant bacteria. These peptides are promising candidates for the development of novel antibacterial agents.


Is there a "net generation" in veterinary medicine? A comparative study on the use of the Internet and Web 2.0 by students and the veterinary profession.
Informal and formal lifelong learning is essential at university and in the workplace. Apart from classical learning techniques, Web 2.0 tools can be used. It is controversial whether there is a so-called net generation amongst people under 30. To test the hypothesis that a net generation among students and young veterinarians exists. An online survey of students and veterinarians was conducted in the German-speaking countries which was advertised via online media and traditional print media. 1780 people took part in the survey. Students and veterinarians have different usage patterns regarding social networks (91.9% vs. 69%) and IM (55.9% vs. 24.5%). All tools were predominantly used passively and in private, to a lesser extent also professionally and for studying. The use of Web 2.0 tools is useful, however, teaching information and media skills, preparing codes of conduct for the internet and verification of user generated content is essential.


Modeling the Cost-Effectiveness of Interventions to Reduce Suicide Risk Among Hospital Emergency Department Patients.
This study estimated the expected cost-effectiveness and population impact of outpatient interventions to reduce suicide risk among patients presenting to general hospital emergency departments (EDs), compared with usual care. Several such interventions have been found efficacious, but none is yet widespread, and the cost-effectiveness of population-based implementation is unknown. Modeled cost-effectiveness analysis compared three ED-initiated suicide prevention interventions previously found to be efficacious-follow-up via postcards or caring letters, follow-up via telephone outreach, and suicide-focused cognitive-behavioral therapy (CBT)-with usual care. Primary outcomes were treatment costs, suicides, and life-years saved, evaluated over the year after the index ED visit. Compared with usual care, adding postcards improved outcomes and reduced costs. Adding telephone outreach and suicide-focused CBT, respectively, improved outcomes at a mean incremental cost of $4,300 and $18,800 per life-year saved, respectively. Monte Carlo simulation (1,000 repetitions) revealed the chance of incremental cost-effectiveness to be a certainty for all three interventions, assuming societal willingness to pay ≥$50,000 per life-year. These main findings were robust to various sensitivity analyses, including conservative assumptions about effect size and incremental costs. Population impact was limited by low sensitivity of detecting ED patients' suicide risk, and health care delivery inefficiencies. The highly favorable cost-effectiveness found for each outpatient intervention provides a strong basis for widespread implementation of any or all of the interventions. The estimated population benefits of doing so would be enhanced by increasing the sensitivity of suicide risk detection among individuals presenting to general hospital EDs.


Binding of correolide to K(v)1 family potassium channels. Mapping the domains of high affinity interaction.
Correolide, a novel nortriterpene natural product, potently inhibits the voltage-gated potassium channel, K(v)1.3, and [(3)H]dihydrocorreolide (diTC) binds with high affinity (K(d) approximately 10 nM) to membranes from Chinese hamster ovary cells that express K(v)1.3 (Felix, J. P., Bugianesi, R. M., Schmalhofer, W. A., Borris, R., Goetz, M. A., Hensens, O. D., Bao, J.-M., Kayser, F. , Parsons, W. H., Rupprecht, K., Garcia, M. L., Kaczorowski, G. J., and Slaughter, R. S. (1999) Biochemistry 38, 4922-4930). Mutagenesis studies were used to localize the diTC binding site and to design a high affinity receptor in the diTC-insensitive channel, K(v)3.2. Transferring the pore from K(v)1.3 to K(v)3.2 produces a chimera that binds peptidyl inhibitors of K(v)1.3 with high affinity, but not diTC. Transfer of the S(5) region of K(v)1.3 to K(v)3.2 reconstitutes diTC binding at 4-fold lower affinity as compared with K(v)1.3, whereas transfer of the entire S(5)-S(6) domain results in a normal K(v)1.3 phenotype. Substitutions in S(5)-S(6) of K(v)1.3 with nonconserved residues from K(v)3.2 has identified two positions in S(5) and one in S(6) that cause significant alterations in diTC binding. High affinity diTC binding can be conferred to K(v)3.2 after substitution of these three residues with the corresponding amino acids found in K(v)1.3. These results suggest that lack of sensitivity of K(v)3.2 to diTC is a consequence of the presence of Phe(382) and Ile(387) in S(5), and Met(458) in S(6). Inspection of K(v)1.1-1.6 channels indicates that they all possess identical S(5) and S(6) domains. As expected, diTC binds with high affinity (K(d) values 7-21 nM) to each of these homotetrameric channels. However, the kinetics of binding are fastest with K(v)1.3 and K(v)1.4, suggesting that conformations associated with C-type inactivation will facilitate entry and exit of diTC at its binding site. Taken together, these findings identify K(v)1 channel regions necessary for high affinity diTC binding, as well as, reveal a channel conformation that markedly influences the rate of binding of this ligand.


Analysis of organo-silica interactions during valve formation in synchronously growing cells of the diatom Navicula pelliculosa.
Biologically formed silica is produced at ambient conditions under the control of molecular and physicochemical processes that are apparently integrated in biosilica morphogenesis, but the mechanisms are not yet fully understood. With the recent identification of small polypeptides and proteins that are encapsulated inside the biosilica and functional in silica polymerization in vitro, it is of importance to determine whether interactions between inorganic silica species and these organic compounds occur in vivo. A time-resolved analysis of valve formation in synchronously growing cells of the diatom species Navicula pelliculosa enabled us to characterize the relevant chemical bonds by attenuated total reflectance Fourier-transformed infrared (ATR-FTIR) spectroscopy. Typically, inorganic bonds of Si-O-Si (bands at 1058, 843 cm(-1)), Si-OH (3689 cm(-1)), and P=O (1239 cm(-1)) and organic bonds of proteinaceous matter (with the amide I and II bands at 1642 and 1543 cm(-1), respectively) were positively identified during one cycle of valve formation. The observed variations in FTIR band intensity and location represented specific interactions between organic and inorganic molecules during the major silicification event, during which stretching of the Si-O bonds was predominantly noticed. The experimentally obtained frequencies (nu) of the major bonds corresponded to those that were obtained by MM+ and PM3 FTIR simulations for organo-silica interactions based on biomolecules that are proposed to be involved in biosilica formation. The results indicated that hydrogen bonds originated from interactions, albeit weak, between organic phosphate or amine groups to the inorganic hydroxyl groups or oxygen atoms from the silicic acid and/or silica. The existence of covalent P-O-Si bonds and electrostatic interactions could not be excluded. These interactions clearly suggest that biomolecules actively contribute to the silica polymerization process during valve formation in N. pelliculosa, and also might act comparably in other diatoms species in which similar biomolecules have been identified.


Sex and age differences in attention-deficit/hyperactivity disorder symptoms and diagnoses: implications for DSM-V and ICD-11.
To examine gender and age differences in attention-deficit/hyperactivity disorder (ADHD) symptom endorsement in a large community-based sample. Families with four or more full siblings ascertained from Missouri birth records completed telephone interviews regarding lifetime DSM-IV ADHD symptoms and the Strengths and Weaknesses of ADHD-Symptoms and Normal-behavior (SWAN) questionnaire for current ADHD symptoms. Complete data were available for 9,380 subjects aged 7 through 29 years. Lifetime and current DSM-IV-like ADHD diagnoses were assigned by the DSM-IV symptom criteria. Linear regression was used to examine sex and age effects on SWAN ADHD symptom scores. Logistic regression was used to examine sex and age effects on specific ADHD diagnoses. Fractional polynomial graphs were used to examine ADHD symptom count variations across age. Overall prevalence of current DSM-IV-like ADHD was 9.2% with a male:female ratio of 2.28:1. The prevalence of DSM-IV-like ADHD was highest in children. Gender differences in DSM-IV-like ADHD subtype prevalences were highest in adolescents. On average, individuals with lifetime DSM-IV-like ADHD diagnoses had elevated current ADHD symptoms even as adolescents or adults. Lower male:female ratios than reported in some clinic-based studies suggest that females are underdiagnosed in the community. Although they may no longer meet the full symptom criteria, young adults with a history of lifetime DSM-IV-like ADHD maintain higher levels of ADHD symptoms compared with the general population. The use of age-specific diagnostic criteria should be considered for DSM-V and ICD-11.


The 2004 Eleanor Clarke Slagle Lecture--Time, space, and the kaleidoscopes of occupation.
Our daily round of occupations occurs within time and space. Our understanding of occupation has traditionally viewed time and space as part of the external environment. Patterns of the "when?" and "where?" of occupations can be described. But relating time and space to the internal experience of occupation reflects more of the meaning to individuals and more complex patterns arise. Like the varied bits of glass in the object case of an art kaleidoscope, the multiple elements of occupation interact. Reflected in mirrors of our choice, we find always-changing, complex patterns of daily occupation. The elements of occupation most important for maintaining or regaining health and the mirrors and lens through which we view occupation historically have formed shifting patterns in occupational therapy, patterns of how we view ourselves, our practice, and those to whom we provide our services. We must combine the science and the art of occupational therapy as the metaphor of the kaleidoscope combines both, producing awe and wonder at the result.


[The problem of posttraumatic fat embolism].
The article is devoted to the problem of fat embolism. In 63 patients with combined and isolated traumas the maximum frequency of fat embolism was diagnosed in the investigation of blood serum from the femoral artery. It can be supposed that no isolated injury of the lesser circulation circuit can occur in fat embolism. An investigation of the venous-arterial difference of fat globulemia has shown the main role of the so-called theory of shunting for fat embolism to be doubtful.


Factors involved in access and utilization of adult hearing healthcare: A systematic review.
Hearing loss is a public health concern, yet hearing healthcare disparities exist and influence utilization of rehabilitation services. The objective of this review was to systematically analyze the published literature on motivators, barriers, and compliance factors affecting adult patient access and utilization of hearing rehabilitation healthcare. Pubmed, PsychINFO, CINAHL, and Web of Science were searched for relevant articles. Eligible studies were those containing original, peer-reviewed research in English pertaining to factors affecting adult hearing healthcare access and utilization of hearing aids and cochlear implantation. The search encompassed 1990 to 2015. Two investigators independently reviewed all articles and extracted data. Specific variables regarding access to care and compliance to recommended care were extracted from each study. Thirty articles were reviewed. The factors affecting access and utilization of hearing rehabilitation could be classified into motivators, barriers, and compliance in treatment or device use. The key motivators to seek care include degree of hearing loss, self-efficacy, family support, and self-recognition of hearing loss. The primary barriers to care were financial limitations, stigma of hearing devices, inconvenience, competing chronic health problems, and unrealistic expectations. Compliance is most affected by self-efficacy, education level, and engagement in the rehabilitation process. Accessing hearing healthcare is complicated by multiple factors. Considering the current climate in healthcare policy and legislation toward improved access of care, a deeper understanding of motivators, barriers, and compliance factors can aid in delivery of effective and efficient hearing healthcare. Laryngoscope, 127:1187-1194, 2017.


Did national folic acid fortification reduce socioeconomic and racial disparities in folate status in the US?
The purpose of this study is to determine the impact of the 1998 US Food and Drug Administration folic acid fortification policy on disparities in folate status in the United States. We use repeated cross-sectional data from the U.S. National Health and Nutrition Examination Surveys (NHANES), a nationally representative sample of over 14 000 participants ages 25 and older. We calculate pre-fortification (1991-94) and post-fortification (1999-2002) absolute differences and relative prevalence ratios of low red blood cell (RBC) folate status (<362.6 nmol), by race/ethnicity and income quartile. We also estimate kernel density plots and relative and absolute concentration curves pre- and post-fortification. The excess prevalence of low RBC folate status associated with the lowest income quartile and black race declined by 67% and 48%, respectively, following fortification. Despite these absolute gains, the relative ratio of low folate status increased after fortification for the lowest compared with the highest income groups (from 1.27 to 2.08) and among whites compared with blacks (from 1.64 to 3.75). The effects of the fortification policy highlight the importance of distinguishing absolute from relative differences when evaluating interventions to reduce health disparities. Targeting of high risk populations is likely needed to eliminate remaining folate disparities.


Intradural disc herniation at L5-S1 mimicking an intradural extramedullary spinal tumor: a case report.
Intradural lumbar disc herniation is a rare pathological entity. The pathogenesis of intradural lumbar disc herniation is not known clearly. Intradural disc herniations usually occurred at the L4-L5 levels but have also been reported at other levels. However, intradural disc herniation at L5-S1 is quite rare. There are approximately nine reports in the English literature of intraradicular disc herniation at L5-S1. We described a 61-yr-old man with suspected intradural mass at the level of L5-S1 space. The patient presented with pain in the lower back and both lower legs for 4 months and a sudden exacerbation of the symptoms for 3 days. Gadolinium-enhanced magnetic resonance imaging (MRI) demonstrated a large disc herniation at the L5-S1 level with an intradural component. L5 and S1 laminectomy was performed, and dura was swollen and immobile. Subsequent durotomy was performed and an intradural disc fragment was removed. The patient had full recovery in 3 months. Intradural lumbar disc herniation must be considered in the differential diagnosis of mass lesions in the spinal canal. Contrast-enhanced MRI scans are useful to differentiate a herniated disc from a disc space infection or tumor.


Fast Quasi-Newton Algorithms for Penalized Reconstruction in Emission Tomography and Further Improvements via Preconditioning.
This paper reports on the feasibility of using a quasi-Newton optimization algorithm, limited-memory Broyden-Fletcher-Goldfarb-Shanno with boundary constraints (L-BFGS-B), for penalized image reconstruction problems in emission tomography (ET). For further acceleration, an additional preconditioning technique based on a diagonal approximation of the Hessian was introduced. The convergence rate of L-BFGS-B and the proposed preconditioned algorithm (L-BFGS-B-PC) was evaluated with simulated data with various factors, such as the noise level, penalty type, penalty strength and background level. Data of three 18F-FDG patient acquisitions were also reconstructed. Results showed that the proposed L-BFGS-B-PC outperforms L-BFGS-B in convergence rate for all simulated conditions and the patient data. Based on these results, L-BFGS-B-PC shows promise for clinical application.


Drug abuse and HIV-related pulmonary hypertension: double hit injury.
: Improved survival among HIV-1-infected individuals with the advent of antiretroviral therapy has clearly led to a greater prevalence of noninfectious complications. One of the most devastating sequelae in these individuals is the development of pulmonary arterial hypertension (PAH). Various epidemiological studies suggest worse survival of HIV-PAH patients when compared with other forms of PAH. Given that only a subset and not all HIV-infected individuals develop HIV-PAH, it is suggested that an additional second-hit of genetic or environmental trigger is needed for the development of PAH. In this context, it has been well documented that HIV patients who abuse illicit drugs such as stimulants, opioids, and the like, are more susceptible to develop PAH. In this review, we highlight the studies that support the significance of a double hit of HIV and drug abuse in the incidence of PAH and focus on the research that has been undertaken to unravel the pathobiology and vascular remodeling mechanisms underlying the deleterious synergy between HIV infection and drugs of abuse in orchestrating the development of PAH.


Methodology of a reevaluation of cardiovascular outcomes in the RECORD trial: study design and conduct.
In 2010, after regulatory review of rosiglitazone licensing, the US Food and Drug Administration (FDA) requested a reevaluation of cardiovascular end points in the RECORD trial. Automated screening of the original clinical trial database and manual case report form review were performed to identify all potential cardiovascular and noncardiovascular deaths, and nonfatal myocardial infarction (MI) and stroke events. Search techniques were used to find participants lost to follow-up, and sites were queried for additional source documents. Suspected events underwent blinded adjudication using both original RECORD end point definitions and new FDA end point definitions, before analysis by the Duke Clinical Research Institute. The reevaluation effort included an additional 328 person-years of follow-up. Automated screening identified 396 suspected deaths, 2,052 suspected MIs, and 468 suspected strokes. Manual review of documents by Duke Clinical Research Institute clinical events classification (CEC) coordinators identified an additional 31 suspected deaths, 49 suspected MIs, and 28 suspected strokes. There were 127 CEC queries issued requesting additional information on suspected deaths; 43 were closed with no site response, 61 were closed with a response that no additional data were available, and additional data were received for 23. Seventy CEC queries were issued requesting additional information for suspected MI and stroke events; 31 were closed with no site response, 20 were closed with a response that no additional data were available, and 19 resulted in additional data. Comprehensive procedures were used for rigorous event reascertainment and readjudication in a previously completed open-label, global clinical trial. These procedures used in this unique situation were consistent with other common approaches in the field, were enhanced to address the FDA concerns about the original RECORD trial results, and could be considered by clinical trialists designing event readjudication protocols for drug development programs that have been completed.


Spirulina Supplementation with High-Intensity Interval Training Decreases Adipokines Levels and Cardiovascular Risk Factors in Men with Obesity.
Adiposity, a state characterized by excessive accumulation of body fat, is closely linked to metabolic complications and the secretion of specific adipokines. This study explores the potential of exercise and Spirulina supplementation to mitigate these complications and modulate adipokine release associated with obesity. The primary objective of this investigation was to examine the impact of a 12-week regimen of high-intensity training combined with Spirulina supplementation on adipokine concentrations and lipid profiles in male individuals with obesity (N = 44). The participants were randomly distributed into four groups, each consisting of 11 participants: a control group (CG), a supplement group (SG), a training group (TG), and a training plus supplement group (TSG). The intervention comprised a 12-week treatment involving Spirulina supplementation (6 g capsule daily), a 12-week high-intensity interval training (HIIT) protocol with three sessions per week, or a combined approach. Following the interventions, metabolic parameters, anthropometric measurements, cardiorespiratory indices, and circulating adipokines [CRP, Sema3C, TNF-α, IL-6, MCP1, IL-8] were assessed within 48 h of the before and final training session. Statistical analyses revealed significant differences across all measures among the groups (p < 0.05). Notably, post hoc analyses indicated substantial disparities between the CG and the three interventional groups regarding body weight (p < 0.05). The combined training and supplementation approach led to noteworthy reductions in low-density lipoprotein (LDL), total cholesterol (TC), and triglyceride (TGL) levels (all p < 0.0001), coupled with an elevation in high-density lipoprotein-cholesterol (HDL-C) levels (p = 0.0001). Furthermore, adipokine levels significantly declined in the three intervention groups relative to the CG (p < 0.05). The findings from this 12-week study demonstrate that Spirulina supplementation in conjunction with high-intensity interval training reduced adipokine levels, improved body weight and BMI, and enhanced lipid profiles. This investigation underscores the potential of Spirulina supplementation and high-intensity interval training as a synergistic strategy to ameliorate obesity-related complications and enhance overall cardiometabolic well-being in obese males.


Germline PALB2 Variants and PARP Inhibitors in Endometrial Cancer.
PARP inhibitors are orally administered antineoplastic agents that affect the homologous recombination (HR) repair pathway, and are approved by the FDA for the treatment of ovarian, breast, pancreatic, and prostate cancers. This report presents a case of recurrent endometrial carcinoma occurring in a woman with a germline pathogenic PALB2 whole-exon deletion. This uncommon finding in a patient with endometrial carcinoma provided the opportunity to use a management strategy of PARP inhibition with olaparib, resulting in a prolonged response to treatment; however, disease progression eventually occurred. Further studies are required to elucidate the mechanisms underlying resistance to PARP inhibition, and the potential future treatment options in this setting. Current recommendations for risk management of female carriers of PALB2 variants focus on breast and ovarian cancer risk. This case raises the additional question of a potential role for risk-reducing hysterectomy in female carriers of PALB2 variants.


Haemodynamic-guided fluid administration for the prevention of contrast-induced acute kidney injury: the POSEIDON randomised controlled trial.
The administration of intravenous fluid remains the cornerstone treatment for the prevention of contrast-induced acute kidney injury. However, no well-defined protocols exist to guide fluid administration in this treatment. We aimed to establish the efficacy of a new fluid protocol to prevent contrast-induced acute kidney injury. In this randomised, parallel-group, comparator-controlled, single-blind phase 3 trial, we assessed the efficacy of a new fluid protocol based on the left ventricular end-diastolic pressure for the prevention of contrast-induced acute kidney injury in patients undergoing cardiac catheterisation. The primary outcome was the occurrence of contrast-induced acute kidney injury, which was defined as a greater than 25% or greater than 0·5 mg/dL increase in serum creatinine concentration. Between Oct 10, 2010, and July 17, 2012, 396 patients aged 18 years or older undergoing cardiac catheterisation with an estimated glomerular filtration rate of 60 mL/min per 1·73 m(2) or less and one or more of several risk factors (diabetes mellitus, history of congestive heart failure, hypertension, or age older than 75 years) were randomly allocated in a 1:1 ratio to left ventricular end-diastolic pressure-guided volume expansion (n=196) or the control group (n=200) who received a standard fluid administration protocol. Four computer-generated concealed randomisation schedules, each with permuted block sizes of 4, were used for randomisation, and participants were allocated to the next sequential randomisation number by sealed opaque envelopes. Patients and laboratory personnel were masked to treatment assignment, but the physicians who did the procedures were not masked. Both groups received intravenous 0·9% sodium chloride at 3 mL/kg for 1 h before cardiac catheterisation. Analyses were by intention to treat. Adverse events were assessed at 30 days and 6 months and all such events were classified by staff who were masked to treatment assignment. This trial is registered with ClinicalTrials.gov, number NCT01218828. Contrast-induced acute kidney injury occurred less frequently in patients in the left ventricular end-diastolic pressure-guided group (6·7% [12/178]) than in the control group (16·3% [28/172]; relative risk 0·41, 95% CI 0·22-0·79; p=0·005). Hydration treatment was terminated prematurely because of shortness of breath in three patients in each group. Left ventricular end-diastolic pressure-guided fluid administration seems to be safe and effective in preventing contrast-induced acute kidney injury in patients undergoing cardiac catheterisation. Kaiser Permanente Southern California regional research committee grant.


X-Ray-Induced Growth Dynamics of Luminescent Silver Clusters in Zeolites.
Herein, AlKα X-rays are used to drive the growth of luminescent silver clusters in zeolites. The growth of the silver species is tracked using Auger spectroscopy and fluorescence microscopy, by monitoring the evolution from their ions to luminescent clusters and then metallic, dark nanoparticles. It is shown that the growth rate in different zeolites is determined by the mobility of the silver ions in the framework and that the growth dynamics in calcined samples obeys the Hill-Langmuir equation for noncooperative binding. Comparison of the optical properties of X-ray-grown silver clusters with silver clusters formed by standard heat treatment indicates that the latter have a higher specificity toward the formation of luminescent clusters of a specific (small) nuclearity, whereas the former produce a wide distribution of cluster species as well as larger nanoparticles.


Ultra-High Temperature Treatment of Liquid Infant Formula, Systemic Immunity, and Kidney Development in Preterm Neonates.
Ready-to-feed liquid infant formulas (IFs) are increasingly being used for newborn preterm infants when human milk is unavailable. However, sterilization of liquid IFs by ultra-high temperature (UHT) introduces Maillard reaction products (MRPs) that may negatively affect systemic immune and kidney development. UHT-treated IF without and with prolonged storage (SUHT) are tested against pasteurized IF (PAST) in newborn preterm pigs as a model for preterm infants. After 5 days, blood leukocytes, markers of systemic immunity and inflammation, kidney structure and function are evaluated. No consistent differences between UHT and PAST pigs are observed. However, SUHT increases plasma TNFα and IL-6 and reduces neutrophils and in vitro response to LPS. In SUHT pigs, the immature kidneys show minor upregulation of gene expressions related to inflammation (RAGE, MPO, MMP9) and oxidative stress (CAT, GLO1), together with glomerular mesangial expansion and cell injury. The increased inflammatory status in SUHT pigs appears unrelated to systemic levels of MRPs. SUHT feeding may impair systemic immunity and affect kidney development in preterm newborns. The systemic effects may be induced by local gut inflammatory effects of MRPs. Optimal processing and length of storage are critical for UHT-treated liquid IFs for preterm infants.


Lysophosphatidic acid is a potential mediator of cholestatic pruritus.
Pruritus is a common and disabling symptom in cholestatic disorders. However, its causes remain unknown. We hypothesized that potential pruritogens accumulate in the circulation of cholestatic patients and activate sensory neurons. Cytosolic free calcium ([Ca(2+)](i)) was measured in neuronal cell lines by ratiometric fluorometry upon exposure to serum samples from pruritic patients with intrahepatic cholestasis of pregnancy (ICP), primary biliary cirrhosis (PBC), other cholestatic disorders, and pregnant, healthy, and nonpruritic disease controls. Putative [Ca(2+)](i)-inducing factors in pruritic serum were explored by analytical techniques, including quantification by high-performance liquid chromatography/mass spectroscopy. In mice, scratch activity after intradermal pruritogen injection was quantified using a magnetic device. Transient increases in neuronal [Ca(2+)](i) induced by pruritic PBC and ICP sera were higher than corresponding controls. Lysophosphatidic acid (LPA) could be identified as a major [Ca(2+)](i) agonist in pruritic sera, and LPA concentrations were increased in cholestatic patients with pruritus. LPA injected intradermally into mice induced scratch responses. Autotaxin, the serum enzyme converting lysophosphatidylcholine into LPA, was markedly increased in patients with ICP versus pregnant controls (P < .0001) and cholestatic patients with versus without pruritus (P < .0001). Autotaxin activity correlated with intensity of pruritus (P < .0001), which was not the case for serum bile salts, histamine, tryptase, substance P, or mu-opioids. In patients with PBC who underwent temporary nasobiliary drainage, both itch intensity and autotaxin activity markedly decreased during drainage and returned to preexistent levels after drain removal. We suggest that LPA and autotaxin play a critical role in cholestatic pruritus and may serve as potential targets for future therapeutic interventions.


Participatory science for coastal water quality: freshwater plume mapping and volunteer retention in a randomized informational intervention.
Among the biggest threats to coastal water quality are freshwater discharges. It is difficult to predict the spatial extent of freshwater plumes at marine beaches because processes governing mass transport in the surf zone are complex. Participatory science approaches could facilitate collecting shoreline data, although volunteer sampling campaigns can be challenged by data quality and volunteer retention. The goals of this study were to (1) work with volunteers to estimate safe swimming distances at beaches that receive polluted discharges, and (2) test whether informational feedback to volunteers increased retention. Forty-six volunteers participated over 12 weeks in 2019 by collecting 1452 salinity measurements at beaches near the mouths of two Central California freshwater discharges and completing participation surveys. These measurements resulted in 145 distinct estimates of safe swimming distances (D90), spanning a range of environmental conditions during rainy and dry periods. Median D90s were 150 and 100 m at San Pedro Creek south and north, and 490 and 330 m at San Lorenzo River west and east, respectively. D90 was significantly associated with adjacent freshwater discharge rate at both discharges and tide level at one discharge. On average, the odds of volunteers conducting sampling decreased by 4% (95% CI: 1%, 7%) with each successive week. A randomized intervention providing repeated data feedback via email to volunteers did not affect their retention in the study.


[Unilateral hypochromic verrucous epidermal nevus (author's transl)].
A case of unilateral hypochromic epidermal nevus is presented. A literature search and review of our own autopsy and ultrastructural studies did not permit the classification of this case to previously described genodermatoses. The possible origin of this congenital cutaneous malformation is discussed.


Extreme altitudes during diurnal flights in a nocturnal songbird migrant.
Billions of nocturnally migrating songbirds fly across oceans and deserts on their annual journeys. Using multisensor data loggers, we show that great reed warblers (Acrocephalus arundinaceus) regularly prolong their otherwise strictly nocturnal flights into daytime when crossing the Mediterranean Sea and the Sahara Desert. Unexpectedly, when prolonging their flights, they climbed steeply at dawn, from a mean of 2394 meters above sea level to reach extreme cruising altitudes (mean 5367 and maximum 6267 meters above sea level) during daytime flights. This previously unknown behavior of using exceedingly high flight altitudes when migrating during daytime could be caused by diel variation in ambient temperature, winds, predation, vision range, and solar radiation. Our finding of this notable behavior provides new perspectives on constraints in bird flight and might help to explain the evolution of nocturnal migration.


[Structural characteristics of a population of antigenic variants of the influenza A(H3N2) virus].
The data have been obtained indicating that clone distribution by the antigen avidity in the population of influenza A (H3N2) virus corresponds to normal distribution. The degree of avidity of individual strains is determined by the predominant content of clones with high or low avidity. Virus purification by ultracentrifugation in sucrose density gradient results in increasing the avidity of the preparation as compared with the original allantoic cultures. Defective virions may differ in avidity from intact viral particles of the same strain and in this way affect the "total" avidity of a virus preparation.


Trough ganciclovir concentration as predictor of leukopenia in lung transplant recipients receiving valganciclovir prophylaxis.
Valganciclovir is the first-line agent for Cytomegalovirus prophylaxis after lung transplantation. However, its use is associated with a relatively high risk of hematological toxicity. This study aimed to investigate the relationship between trough ganciclovir concentration and hematologic toxicity in lung transplantation patients receiving valganciclovir prophylaxis, and identify factors that affect ganciclovir pharmacokinetics in this population. This prospective observational study included 24 lung transplant patients receiving valganciclovir prophylaxis. The cutoff value of trough ganciclovir concentration was estimated using receiver operating characteristic analysis in leukopenia grade 3 and higher. Population pharmacokinetic analysis was performed using a nonlinear mixed-effects modeling program. The trough ganciclovir concentration was significantly higher in the group with leukopenia grades 3 or higher than in the group with grades less than or equal to 2 (1605.7 ± 860.1 ng/mL [n = 3] vs. 380.5 ± 175.8 ng/mL (n = 21), p < .001). The cutoff value of trough ganciclovir concentration for predicting greater than or equal to grade 3 leukopenia was estimated as 872.0 ng/mL. Creatinine clearance and lung re-transplantation were found to have a significant impact on the total body clearance of valganciclovir. Ganciclovir clearance was decreased in patients with reduced creatine clearance or re-transplantation. These results suggest that higher ganciclovir trough concentrations are associated with an increased risk of leukopenia grade 3 or higher, and that creatinine clearance and lung re-transplantation affected the pharmacokinetics of ganciclovir.


A comprehensive overview of the Chloroflexota community in wastewater treatment plants worldwide.
Chloroflexota are often abundant members of the biomass in wastewater treatment plants (WWTPs) worldwide, typically with a filamentous morphology, forming the backbones of the activated sludge floc. However, their overgrowth can often cause operational issues connected to poor settling or foaming, impairing effluent quality and increasing operational costs. Despite their importance, few Chloroflexota genera have been characterized so far. Here, we present a comprehensive overview of Chloroflexota abundant in WWTPs worldwide and an in-depth characterization of their morphology, phylogeny, and ecophysiology, obtaining a broad understanding of their ecological role in activated sludge.


Glucose response of dissolved-core alginate microspheres: towards a continuous glucose biosensor.
Microparticle optical sensors hold potential as implantable smart materials for in vivo analysis. In this work, the reversible response of dissolved-core alginate microspheres containing a homogeneous fluorescence resonance energy transfer (FRET)-based competitive binding assay for glucose was evaluated. The layer-by-layer self assembly technique was used to deposit multilayered nanofilm coatings on the alginate microspheres containing the assay, thereby stabilizing the sensor system when the alginate was de-crosslinked. The response to glucose was then determined in DI water and simulated interstitial fluid (SIF) using a flow cell to establish controlled, dynamic flow conditions for demonstrating reversibility. The glucose sensitivity under dynamic conditions was estimated to be 0.52%/mM glucose in DI water and 0.6%/mM glucose in simulated interstitial fluid; in both cases, the analytical response range was 0-30 mM glucose, covering both physiological (normoglycemia) and pathophysiological range (hyperglycemia and hypoglycemia). The sensor demonstrated a repeatable and reproducible response when tested over a period of one month, under dynamic flow conditions. Finally, in vitro cytotoxicity assays performed with L929 mouse fibroblast cell lines suggested that the dissolved-core alginate microsphere sensor system with nanofilm coating has sufficient biocompatibility for use as implantable glucose biosensors.


Mono-aromatic ring-fused versus adjacently di-aromatic ring-fused tetraazaporphyrins: regioselective synthesis and their spectroscopic and electrochemical properties.
A method which preferentially produces adjacently di-aromatic ring-substituted tetraazaporphyrins (TAPs) has been developed, and their electrochemical and spectroscopic properties have been studied and compared with those of the corresponding series of mono-aromatic ring-fused TAPs. Mono-aromatic ring-fused TAPs show a split Q-band, and the splitting energy increases with increasing size of the aromatic ring. In addition, for the split Q-bands, the relative intensity of the band at longer wavelength decreases with increasing molecular size of the fused aromatics, compared with the shorter wavelength band. In the di-aromatic ring-fused TAPs, this kind of splitting is not seen, and only a shift of the band is observed. The intensity and band position of the split or unsplit Q-bands are quantitatively evaluated by simultaneous band deconvolution analysis, using both electronic absorption and magnetic circular dichroism spectra. The preparation of these TAP compounds has made it possible to adjust the Q-band position in a stepwise manner between ca. 600 and 750 nm. The first reduction and oxidation potentials of the TAP ring shift negatively with increasing number and size of the fused aromatics. The extent of the shift is found to be very small for the LUMOs but significant for the HOMOs. These spectroscopic and electrochemical properties are almost perfectly reproduced by molecular orbital calculations within the framework of the Pariser-Parr-Pople approximation. In particular, a small variation of the LUMO level and large destabilization of the HOMO level on ring expansion are rationalized from the extent of stretch of molecular orbitals: i.e., since the LUMOs are localized in the central TAP moiety irrespective of the molecular size, while the HOMOs have appreciable coefficients even over the fused aromatics, the HOMO level destabilizes while the LUMO level remains constant with increasing molecular size. In one CoTAP derivative, Co(III/II) and the first ligand oxidation couples occur experimentally at the same potential.


Disease evidence for IGFBP-2 as a key player in prostate cancer progression and development of osteosclerotic lesions.
Accumulating evidence indicates that alterations in the IGF axis contribute to the development of chemo- and radio-resistant, advanced-stage cancers. Additionally, they contribute to hormonal insensitivity in adenocarcinomas such as those derived from prostate and breast. The ligands, IGF-I and IGF-II, along with their receptors, IGF-IR and IGF-IIR, have been implicated in a wide range of disease. Activation and subsequent signal transduction through the receptors is attenuated, and/or potentiated, by the interactions of IGF axis ligands, IGF-I/II, with the high affinity IGF-binding proteins 1 to 6 (IGFBP1-6). New evidence indicates that the IGFBPs, irrespective of ligand interactions, correlate with the development and metastatic behavior of several cancers. Increased expression of insulin-like growth factor binding protein 2 (IGFBP-2) is found in advanced cancers of the ovary, breast, stomach, adrenal gland, bladder, CNS, and prostate. Further, IGFBP-2 seemingly has ligand-independent effects that participate in the development and dissemination of advanced cancer cells. As such, IGFBP-2 can assist in the development of the lethal phenotype for some cancers. While several reports have shown an important role for IGFBP-2 in the development of androgen insensitivity and the proliferation of AI PCa cells in vivo, these studies have not tested a role for IGFBP-2 in the metastatic spread of AI PCa cells. Additionally, the mechanism of IGFBP-2 action in these events has not been elucidated. The redundancy and abundance of the IGFBPs have precluded a clear understanding of the means by which IGFBP-2 signals. Components of these signaling pathways, particularly IGFBP-2, are being evaluated currently in clinical trials.


[NMR study of the structural transition in the DNA-bound water system in the interval of physiological temperatures].
Temperature dependence of spin-spin proton relaxation times of DNA and bound water and the content of bound water in the samples of DNA, saturated with water in the atmosphere with different relative humidities from 0 to 100% were studied by means of pulsed NMR. It is shown that the temperature transition in the system of DNA-bound water in the interval 18-35 degrees is observed only when the relative humidity is more than 70% and the double-stranded structure of DNA exists. The transition of DNA from one conformation into another passes through some intermediate state more labile and probably less ordered. This transition is accompanied by changes in the structure of the hydration shell. In the case when relative humidity is greater than 80%, the partial dehydration of DNA stimulated by the transition is observed. This dehydration increases with the increase of relative humidity.


Differences in anatomy and outcomes in patients treated with open mesenteric revascularization before and after the endovascular era.
To compare the clinical characteristics, anatomy, and outcomes of patients treated with open mesenteric revascularization (OR) for chronic mesenteric ischemia (CMI) before and after the preferential use of endovascular revascularization (ER). We reviewed a prospective database of 257 patients treated for CMI with OR or ER from 1998 to 2009. Treatment trends were analyzed to identify changes in practice paradigm. Prior to 2002, OR was used in 58 of 81 patients (72%). Since 2002, ER surpassed OR as the most common treatment option; OR was indicated in 58 of 176 patients (33%) who either failed ER or had unfavorable lesions for stent placement. We analyzed differences in clinical data, anatomical characteristics, and outcomes in 116 patients treated with OR before (Pre-Endo, n = 58) and after 2002 (Post-Endo, n = 58). Anatomical characteristics were determined by a blinded investigator using conventional angiography, magnetic resonance angiography, and computed tomography angiography with centerline of flow measurements. Both groups had similar demographics, risk factors, and clinical presentation, with the exception of higher (P < .05) rates of hypertension, hyperlipidemia, cardiac interventions, dysrhythmias, and higher comorbidity scores in the Post-Endo group. This group also had more extensive mesenteric artery disease, including higher incidence of three-vessel involvement (76% vs 57%; P = .048) and superior mesenteric artery (SMA) occlusion (67% vs 41%;P = .005). There were no differences (P > .05) in the number of vessels revascularized (1.8 ± 0.4 vs 1.7 ± 0.5) and in graft configuration (antegrade, 91% vs 78%; retrograde, 9% vs 22%; two-vessel, 69% vs 81%) in the Pre- and Post-Endo groups, respectively. There were no differences in operative mortality (1.7% vs 3.4%), morbidity (43% vs 53%), length of stay (12 ± 1 vs 12 ± 1 days), and immediate symptom improvement (88% vs 86%) in the Pre- and Post-Endo groups, respectively. Mean follow-up was 57 ± 6 months for patients treated before 2002 and 29 ± 6 months for those treated after 2002 (P = .0001). At 5 years, primary and secondary patency rates and recurrence-free survival were 82%, 86%, and 84% in the Pre-Endo and 81%, 82%, and 76% in the Post-Endo groups (P > .05). OR has been used in approximately one-third of patients treated for CMI since 2002. Despite more comorbidities and more extensive mesenteric artery disease in patients now treated with OR, outcomes have not changed compared with those operated prior to the preferential use of mesenteric stents before 2002.


Appropriateness of antimicrobial therapy: a multicentre prevalence survey in the Netherlands, 2008-2009.
A survey was carried out to determine the prevalence and appropriateness of antimicrobial therapy (AMT) in the Netherlands and to identify determinants for inappropriate AMT. Prevalence surveys of patients hospitalised in the Netherlands were performed three times in 2008 and 2009. Patients' demographic, infection-related and AMT-related data were collected from hospital wards. A total of 19 hospitals participated, consisting of a mix of university, teaching and general hospitals, which were distributed evenly across the country. The appropriateness of AMT was assessed using a standardised algorithm based on local AMT prescription guidelines. A total of 7,853 patients were included, of which 2,327 (29.6%) patients were on AMT (range: 20.8–39.5%). In 372 patients (16% of patients on AMT), treatment was considered inappropriate. In 265 (11.4%) patients on AMT, appropriateness of treatment was not judged because of insufficient information. The percentage of patients without a judgment varied considerably between the participating hospitals (range: 1.3–36.2%). Appropriate AMT use was significantly associated with a patient being in an intensive care unit, having a central venous catheter and being given beta-lactamase-sensitive penicillins. The use of fluoroquinolones was significantly associated with more frequent inappropriate use. There was considerable and significant variation between the participating hospitals in the amount of antimicrobials prescribed and the appropriateness of their use. To improve the completeness and reliability of such surveys, there is a need for intensive training of observers and medical staff in recording information.


Clinical engineering helps reduce equipment costs.
Clinical engineering involves, among other activities, managing equipment repair and maintenance activities, assessing technology needs and potential equipment acquisitions, training users, negotiating service contracts, and optimizing patient-care equipment utilization. Effective clinical engineering programs can help healthcare organizations reduce the high cost of acquiring and maintaining patient care equipment.


Expression of insulin-like growth factor system components in colorectal tissue and its relation with serum IGF levels.
The insulin-like growth factor (IGF)-system has been implicated in colorectal tumor carcinogenesis. Although both tumor expression levels and serum concentrations of IGF-system components are related to colorectal cancer risk, it is unknown whether IGF levels in tissue and serum are correlated. The objective of this study was to determine expression levels of various IGF-system components in different locations of the colorectum, and to investigate whether normal tissue IGF expression levels are correlated with serum IGF-I and IGF-II concentrations. Biopsies from macroscopically normal mucosa at four locations in the colorectum (ascending, transverse, sigmoid colon, and rectum) and a fasting serum sample were obtained from 48 asymptomatic patients at increased risk of colorectal cancer. Expression levels of IGF-I, IGF-II, IGF-IR, IGF-IIR, and IGFBP-3 messenger RNA (mRNA) in tissue were quantitatively evaluated using real-time RT-PCR. Expression of IGF-IR protein in the ascending colon and rectum tissue specimens was assessed semi-quantitatively by immunohistochemistry. Serum IGF-I and IGF-II concentrations were determined using immunometric assays. With the exception of IGF-IIR, mRNA levels of all the IGF-system components investigated, as well as IGF-IR protein expression, were significantly higher in the rectum compared with the ascending colon (p<or=0.001). Serum IGF-I and IGF-II concentrations did not correlate with any of the parameters studied in colorectal tissues. Our results indicate that in humans IGF-system components are differentially expressed in the colorectum. Moreover, our findings suggest that local and circulating components of the IGF-system are differentially regulated. However, due to large intra-individual variation in mRNA expression, we cannot formally exclude undetected but existing routes of co-regulation.


[The hemolytic-uremic syndrome].
The microangiopathic thrombotic syndromes--thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS)--are characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal dysfunction, fever and central nervous system abnormalities. Today they are considered as two extremes of a continuous spectrum named TTP--HUS. The syndrome is an uncommon disease with a high mortality rate, despite treatment. The authors describe a case of hemolytic uremic syndrome in a young adult patient. Initially the clinical course and the first biopsy suggested a favourable prognosis, but the early recurrence with severe hypertension was followed by a fatal outcome 6 months later. Concerning this clinical case, the authors present a review of the most recent aspects of the pathogenesis and treatment of this syndrome.


Top 10 research questions related to assessing physical activity and its contexts using systematic observation.
Numerous methods are available to assess physical activity (PA) but systematic observation (SO) excels in being able to provide contextually rich data on the setting in which the activity occurs. As SO is particularly useful for determining how activity is influenced by the immediate physical and social environments, its use is becoming more popular. Observation tools have the advantages of flexibility, high internal validity, low inference, and low participant burden, while their disadvantages include the need for careful observer training and recalibration, inaccessibility to certain environments, and potential participant reactivity. There is a need for both scientists and practitioners to have additional information on observation techniques and systems relative to making environmental and policy decisions about PA, and in this article, we describe concepts and identify questions related to using SO in researching PA behavior. We present 10 general questions in 3 sections, including those related to: (a) ensuring data accuracy through the selection of the most appropriate methodological protocols; (b) investigating PA in school settings, including physical education, recess, and other programs; and (c) investigating PA in community settings (e.g., parks, recreation centers, youth and adult sport programs) and homes.


Simultaneous expression of neuronal and glial properties by chick ciliary ganglion cells during development.
Autoradiographic methods were used to show that non-neuronal cells from dissociated chick ciliary ganglia grown in cell culture for 1 day exhibit high affinity uptake for norepinephrine (NE) and/or specific receptors for nerve growth factor (NGF). Using immunofluorescence procedures, it was demonstrated that these cells reacted neither with the neuron-specific marker tetanus toxin nor with antibodies to the fibroblast marker fibronectin. The cells were, however, positive for 04 antigen, which is present on Schwann cells and oligodendrocytes and is recognized by a monoclonal antibody (Schachner, M., S. K. KIm, and R. Zehnle (1981) Dev. Biol. 83: 328-338). At all stages studied between embryonic day 6 (E6) and embryonic day 14 (E14), about 80% of the non-neuronal cells were positive for 04 antigen, the other non-neuronal cells being identified as fibroblasts or fibroblast-like cells by staining with antibodies to fibronectin. The proportion of cells with NGF receptors and cells with NE uptake decreased during development between E6 and E14. The percentage of 04-positive cells which have NGF receptors decreased from about 95% at E6 to about 35% at E14. The proportion of 04-positive cells with NE uptake decreased from about 57% at E6 to about 15% at E14. Thus, a considerable proportion of the non-neuronal cell population in embryonic ciliary ganglia displays neuronal properties. We suggest that those cells exhibiting biochemical properties of both differentiated glial cells and neurons are precursor cells which have the potential to develop either into glial cells or neurons.


Axin utilizes distinct regions for competitive MEKK1 and MEKK4 binding and JNK activation.
Axin is a multidomain protein that plays a critical role in Wnt signaling, serving as a scaffold for down-regulation of beta-catenin. It also activates the JNK mitogen-activated protein kinase by binding to MEKK1. However, it is intriguing that Axin requires several additional elements for JNK activation, including a requirement for homodimerization, sumoylation at the extreme C-terminal sites, and a region in the protein phosphatase 2A-binding domain. In our present study, we have shown that another MEKK family member, MEKK4, also binds to Axin in vivo and mediates Axin-induced JNK activation. Surprisingly MEKK4 binds to a region distinct from the MEKK1-binding site. Dominant negative mutant of MEKK4 attenuates the JNK activation by Axin. Activation of JNK by Axin in MEKK1-/- mouse embryonic fibroblast cells supports the idea that another MEKK can mediate Axin-induced JNK activation. Expression of specific small interfering RNA against MEKK4 effectively attenuates JNK activation by the MEKK1 binding-defective Axin mutant in 293T cells and inhibits JNK activation by wild-type Axin in MEKK1-/- cells, confirming that MEKK4 is indeed another mitogen-activated protein kinase kinase kinase that is specifically involved in Axin-mediated JNK activation independently of MEKK1. We have also identified an additional domain between MEKK1- and MEKK4-binding sites as being required for JNK activation by Axin. MEKK1 and MEKK4 compete for Axin binding even though they bind to sites far apart, suggesting that Axin may selectively bind to MEKK1 or MEKK4 depending on distinct signals or cellular context. Our findings will provide new insights into how scaffold proteins mediate ultimate activation of different mitogen-activated protein kinase kinase kinases.


SUMOylation in Human Pathogenic Fungi: Role in Physiology and Virulence.
The small ubiquitin-related modifier (SUMO) protein is an important component of the post-translational protein modification systems in eukaryotic cells. It is known to modify hundreds of proteins involved in diverse cellular processes, ranging from nuclear pore dynamics to signal transduction pathways. Owing to its reversible nature, the SUMO-conjugation of proteins (SUMOylation) holds a prominent place among mechanisms that regulate the functions of a wide array of cellular proteins. The dysfunctional SUMOylation system has been associated with many human diseases, including neurodegenerative and autoimmune disorders. Furthermore, the non-pathogenic yeast Saccharomyces cerevisiae has served as an excellent model to advance our understanding of enzymes involved in SUMOylation and proteins modified by SUMOylation. Taking advantage of the tools and knowledge obtained from the S. cerevisiae SUMOylation system, research on fungal SUMOylation is beginning to gather pace, and new insights into the role of SUMOylation in the pathobiology of medically important fungi are emerging. Here, we summarize the known information on components of the SUMOylation machinery, and consequences of overexpression or deletion of these components in the human pathogenic fungi, with major focus on two prevalent Candida bloodstream pathogens, C. albicans and C. glabrata. Additionally, we have identified SUMOylation components, through in silico analysis, in four medically relevant fungi, and compared their sequence similarity with S. cerevisiae counterparts. SUMOylation modulates the virulence of C. albicans and C. glabrata, while it is required for conidia production in Aspergillus nidulans and A. flavus. In addition to highlighting these recent developments, we discuss how SUMOylation fine tunes the expression of virulence factors, and influences survival of fungal cells under diverse stresses in vitro and in the mammalian host.