Effects of cigarette smoking on Fas/Fas ligand expression of human lymphocytes.
Cigarette smoking has been shown to affect human immune responses. We have studied Fas/Fas ligand (FasL) expression, which is involved in the cytotoxic activity, immune privilege, and self-tolerance, and other apoptosis-associated molecule expression of peripheral blood lymphocytes (PBL) in healthy subjects with/without cigarette smoking. We found that expression of FasL protein was detected marginally in the fresh PBL and was induced upon mitogen activation in normal individuals without smoking. In contrast, fresh PBL from those with chronic cigarette smoking exhibited enhanced expression of FasL protein without in vitro mitogen stimulation. Moreover, mitogen stimulation failed to augment FasL protein expression of their lymphocytes, suggesting dysregulation of FasL expression of PBL in individuals with cigarette smoking. In contrast, Fas, Bcl-2, and p53 expression were not significantly different between normal individuals with chronic cigarette smoking and those without smoking. In addition, we found that in vitro brief treatment with nicotine induces and/or enhances FasL mRNA and protein expression of lymphocytes from normal donors without smoking. These results suggest that aberrant FasL expression of lymphocytes is, at least in part, involved in the immune impairment in individuals with chronic cigarette smoking.


Transfer of multidrug resistance among acute myeloid leukemia cells via extracellular vesicles and their microRNA cargo.
The treatment of acute leukemia is still challenging due in part to the development of resistance and relapse. This chemotherapeutics resistance is established by clonal selection of resistant variants of the cancer cells. Recently, a horizontal transfer of chemo-resistance among cancer cells via extracellular vesicles (EVs) has been suggested. The aim of this research was to investigate the role of EVs in chemo-resistance in acute myeloid leukemia. For this purpose, the sensitive strain of the promyelocytic leukemia HL60 cell line was studied along with its multi-resistant strain, HL60/AR that overexpresses the multidrug resistance protein 1 (MRP-1). A chemo-resistance transfer between the two strains was established by treating HL60 cells with EVs generated by HL60/AR. This study reveals that EVs from HL60/AR can interact with HL60 cells and transfer at least partially, their chemo-resistance. EVs-treated cells begin to express MRP-1 probably due to a direct transfer of MRP-1 and nucleic acids transported by EVs. In this context, two microRNAs were highlighted for their high differential expression in EVs related to sensitive or chemo-resistant cells: miR-19b and miR-20a. Because circulating microRNAs are found in all biological fluids, these results bring out their potential clinical use as chemo-resistance biomarkers in acute myeloid leukemia.


Exercise training does not correct abnormal cardiac glycogen accumulation in the db/db mouse model of type 2 diabetes.
Substrate imbalance is a well-recognized feature of diabetic cardiomyopathy. Insulin resistance effectively limits carbohydrate oxidation, resulting in abnormal cardiac glycogen accumulation. Aims of the present study were to 1) characterize the role of glycogen-associated proteins involved in excessive glycogen accumulation in type 2 diabetic hearts and 2) determine if exercise training can attenuate abnormal cardiac glycogen accumulation. Control (db(+)) and genetically diabetic (db/db) C57BL/KsJ-lepr(db)/lepr(db) mice were subjected to sedentary or treadmill exercise regimens. Exercise training consisted of high-intensity/short-duration (10 days) and low-intensity/long-duration (6 wk) protocols. Glycogen levels were elevated by 35-50% in db/db hearts. Exercise training further increased (2- to 3-fold) glycogen levels in db/db hearts. Analysis of soluble and insoluble glycogen pools revealed no differential accumulation of one glycogen subspecies. Phosphorylation (Ser(640)) of glycogen synthase, an indicator of enzymatic fractional activity, was greater in db/db mice subjected to sedentary and exercise regimens. Elevated glycogen levels were accompanied by decreased phosphorylation (Thr(172)) of 5'-AMP-activated kinase and phosphorylation (Ser(79)) of its downstream substrate acetyl-CoA carboxylase. Glycogen concentration was not associated with increases in other glycogen-associated proteins, including malin and laforin. Novel observations show that exercise training does not correct diabetes-induced elevations in cardiac glycogen but, rather, precipitates further accumulation.


Depression and preference for self-focusing stimuli after success and failure.
Depressed individuals, who tend to have large perceived-self/ideal-self discrepancies, have been shown to be particularly high in private self-consciousness. On the bases of this finding and of several converging theoretical perspectives, we hypothesized that depressives, unlike nondepressives, do not find self-focus more aversive after failure than after success, and thus either (a) show no differential preference for self-focusing stimuli after success versus after failure (weak hypothesis), or (b) prefer self-focusing stimuli after failure over self-focusing stimuli after success (strong hypothesis). Depressed and nondepressed college students succeeded or failed on a supposed test of verbal intelligence, and then worked on two sets of puzzles, one in the presence and one in the absence of a self-focusing stimulus (mirror). Whereas nondepressed subjects liked the mirror-associated puzzle more after success than after failure, depressed subjects did not; depressed subjects tended to like the mirror-associated puzzle more after failure than after success. Nondepressed subjects also exhibited a self-serving pattern of attributions, viewing the test as less valid and luck as more responsible for their performance after failure than after success; depressed subjects showed no such differences. In consistency with their failure to use defensive strategies, depressed subjects showed a decrease in self-esteem after failure; nondepressed subjects showed no such change.


High Degree of NCAM Diversity Generated by Alternative RNA Splicing in Brain and Muscle.
The neural cell adhesion molecules (NCAMs) are cell surface glycoproteins involved in vertebrate cell contact formation. Several NCAM mRNA types are generated from a single primary transcript by alternative splicing and differential polyadenylation. In this presentation we analyse sequence heterogeneities within NCAM transcripts detected in the junctions of exons 7/8, 12/13 and 13/14. The highest degree of evolutionary conservation is observed in the 30-nucleotide insertion (pi) between exons 7 and 8, coding for an identical peptide sequence in the mouse, rat and chicken. The most complex splicing pattern is found between exons 12 and 13, called splice site a. Three alternative exons of 15, 48 and 42 nucleotides can be inserted in various combinations, which may also contain the additional trinucleotide AAG. In mouse muscle cell lines, differential 'extra exon' and AAG usage in splice site a creates up to 16 NCAM diversity forms, some (if not all) of which are also expressed in mouse brain. Additional microdiversity is generated by the insertion of an alternative AAG trinucleotide in exon junction 13/14. If all combinations of splicing patterns identified so far were to occur and to be translated, there could be up to 192 different NCAM proteins.


The excretion and stability to metabolism of bretylium.
Bretylium (o-bromobenzylethyldimethylammonium) is a new type of hypotensive drug. It was estimated in extracts of human urine as the methyl orange complex. From 7 to 45% of a single oral dose was found in human urine within 9 hr. The [(14)C] labelled drug was used to investigate excretion by cats. A minor proportion of a subcutaneous dose was eliminated by cats in the faeces, probably after secretion into the bile. Most of the dose was excreted unchanged in the urine. No products of metabolism were found in either human or cat urine. The drug suffered negligible change when incubated with rat liver tissue in vitro.


Differential valine metabolism in adipose tissue of low and high fat-oxidizing obese subjects.
Differences in fat metabolism are of importance in relation to energy balance. Low fat-oxidizers (LFO) are thought to be more prone for developing obesity. We studied whether LFO have different fasting adipose tissue (AT) protein profiles than high fat-oxidizers (HFO). Six LFO and six HFO subjects were selected from an obese group (n = 99, body mass index>30 kg/m(2) ) taking part in a multi-center study (Nutrient-Gene interaction in human obesity) based on the postprandial fat oxidation capacity after a high fat load. AT protein profiles were studied by 2-DE. Differential proteins were clustered with MAPPfinder according to their function. Protein profiles of purified blood cells and adipocytes served to confine the comparison to adipocyte-specific proteins in AT profiles of LFO and HFO subjects. LFO had increased mitochondrial ROS scavengers possibly related to long-chain unsaturated fatty acid-induced increases in mitochondrial ROS-production. Carbohydrate oxidation seemed to be reduced since expression of several proteins from the glycolysis pathway was lower in LFO. Up-regulation of the valine catabolism at the level of methylmalonate-semialdehyde dehydrogenase appeared to be (part of) the compensatory mechanism. In conclusion, the fasting AT protein profile of LFO and HFO differ at the level of ROS scavenging, the glycolysis pathway and valine metabolism.


Mtsym6, a gene conditioning Sinorhizobium strain-specific nitrogen fixation in Medicago truncatula.
The availability of a wide range of independent lines for the annual medic Medicago truncatula led us to search for natural variants in the symbiotic association with Sinorhizobium meliloti. Two homozygous lines, Jemalong 6 and DZA315.16, originating from an Australian cultivar and a natural Algerian population, respectively, were inoculated with two wild-type strains of S. meliloti, RCR2011 and A145. Both plant lines formed nitrogen-fixing (effective) nodules with the RCR2011 strain. However, the A145 strain revealed a nitrogen fixation polymorphism, establishing an effective symbiosis (Nod(+)Fix(+)) with DZA315.16, whereas only small, white, non-nitrogen fixing nodules (Nod(+)Fix(-)) were elicited on Jemalong 6. Cytological studies demonstrated that these non-fixing nodules are encircled by an endodermis at late stages of development, with no visible meristem, and contain hypertrophied and autofluorescent infection threads, suggesting the induction of plant defense reactions. The non-fixing phenotype is independent of growth conditions and determined by a single recessive allele (Mtsym6), which is located on linkage group 8.


The Shared Ethical Responsibility of Medically and Non-medically Qualified Experts in Human Drug Development Teams.
The complexity of developing and applying increasingly sophisticated new medicinal products has led to the participation of many non-medically qualified scientists in multi-disciplinary non-clinical and clinical drug development teams world-wide. In this introductory paper to the "IFAPP International Ethics Framework for Pharmaceutical Physicians and Medicines Development Scientists" it is argued that all members of such multidisciplinary teams must share the scientific and ethical responsibilities since they all influence directly or indirectly both the outcome of the various phases of the medicines development projects and the safety of the research subjects involved. The participating medical practitioner retains the overriding responsibility and the final decision to stop a trial if the well-being of the research subjects is seriously endangered. All the team members should follow the main ethical principles governing human research, the respect for autonomy, justice, beneficence and non-maleficence. Nevertheless, the weighing of these principles might be different under various conditions according to the specialty of the members.


Patient preferences, knowledge and beliefs about kidney allocation: qualitative findings from the UK-wide ATTOM programme.
To explore how patients who are wait-listed for or who have received a kidney transplant understand the current UK kidney allocation system, and their views on ways to allocate kidneys in the future. Qualitative study using semistructured interviews and thematic analysis based on a pragmatic approach. 10 deceased-donor kidney transplant recipients, 10 live-donor kidney transplant recipients, 12 participants currently wait-listed for a kidney transplant and 4 participants whose kidney transplant failed. Semistructured telephone interviews conducted with participants in their own homes across the UK. Three main themes were identified: uncertainty of knowledge of the allocation scheme; evaluation of the system and participant suggestions for future allocation schemes. Most participants identified human leucocyte anitgen matching as a factor in determining kidney allocation, but were often uncertain of the accuracy of their knowledge. In the absence of information that would allow a full assessment, the majority of participants consider that the current system is effective. A minority of participants were concerned about the perceived lack of transparency of the general decision-making processes within the scheme. Most participants felt that people who are younger and those better matched to the donor kidney should be prioritised for kidney allocation, but in contrast to the current scheme, less priority was considered appropriate for longer waiting patients. Some non-medical themes were also discussed, such as whether parents of dependent children should be prioritised for allocation, and whether patients with substance abuse problems be deprioritised. Our participants held differing views about the most important factors for kidney allocation, some of which were in contrast to the current scheme. Patient participation in reviewing future allocation policies will provide insight as to what is considered acceptable to patients and inform healthcare staff of the kinds of information patients would find most useful.


Microsomal cytochrome P450 and eicosanoid metabolism.
The demonstration of a role for microsomal P450 in the metabolism of endogenous pools of arachidonic acid established this enzyme system as a member of the arachidonic acid cascade and characterized a new an important metabolic function for this enzyme system. Studies from several laboratories documenting the powerful biological activities of the P450-derived eicosanoids have suggested important roles for the P450 arachidonic acid monooxygenase in renal and vascular physiology, and in the pathophysiology of experimental hypertension. These studies provide significant evidence to indicate that in addition to its recognized traditional toxicological and pharmacological roles, microsomal P450s also play important physiological roles in the control of tissue and body homeostasis.


Experimental results of a residential house fire test on tenability: temperature, smoke, and gas analyses.
A fire experiment conducted in a British 1950s-style house is described. Measurements of temperature, smoke, CO, CO2 , and O2 were taken in the Lounge, stairwell, and front and back bedrooms. The front bedroom door was wedged open, while the door to the back bedroom was wedged closed. Contrary to expectations and despite the relatively small fire load, analysis and hazard calculations show permeation of toxic fire gases throughout the property with lethal concentrations of effluent being measured at each sampling point. A generally poor state of repair and missing carpets in the upper story contributed to a high degree of gas and smoke permeation. The available egress time was calculated as the time before the main escape route became impassable. Given known human responses to fire, such an incident could have caused fatalities to sleeping or otherwise immobile occupants.


Aggressive chemotherapy for diffuse histiocytic lymphoma in the elderly: increased complications with advancing age.
Twenty patients greater than or equal to 70 years of age were included in a study of the treatment of diffuse histiocytic lymphoma utilizing cyclophosphamide, adriamycin, vincristine, and prednisone. These patients ranged in age from 70 to 94 years (median 75 years). There were also 55 younger patients (age range 33 to 69 years) in the treatment trial. There were no dose adjustments for age. The complete remission rate in the elderly patients (45 per cent) was not different from that in the younger patients (53 per cent). The overall survival in the elderly patients (median 13 months) was somewhat shorter than that in the younger patients (medians 22 months for patients 56-69 years of age and 41 months for patients 33-55 years of age), but not significantly different. Death during the first two treatment cycles from causes other than lymphoma occurred in 25 per cent of the patients greater than or equal to 70 years of age versus 2 per cent of younger patients (P less than 0.01). In addition, three other patients aged 79, 65, and 59 years died in the fourth or fifth cycles of treatment from causes other than lymphoma. Thus, 30 per cent of patients greater than or equal to 70 years of age died during therapy from causes other than lymphoma, versus 5 per cent of younger patients (P less than 0.01). Whether this altered ability to tolerate therapy in the older patients reflected decreased marrow function, altered drug metabolism, other effects of aging, or a combination of these factors is not clear. It might be appropriate to alter drug doses when treating elderly patients, and particular attention to supportive measures seems appropriate.


Spectrum of developmental disabilities: continuum of motor dysfunction.
The spectrum of developmental disabilities encompasses a diverse group of interrelated disorders with multiple manifestations resulting from brain dysfunction. Motor disability has been highlighted, as it is the most frequent presenting sign of developmental disability. Such disability may vary from severe to minimal. Severe disability, or cerebral palsy, is well appreciated, as are the additional associated disabilities that affect habilitation. Mild to minimal motor disability (mild to minimal cerebral palsy), because it is usually nonhandicapping, is less frequently appreciated as a marker of brain dysfunction. Further delineation of the child's nonmotor abilities is necessary in order to define the scope of the disability. Cognitive limitation is not an important cause for the gross motor delay seen in children with mental retardation. It is our postulation that the motor deficit is on the basis of mild to minimal cerebral palsy. This diagnosis is frequently obscured by the mental retardation.


Impact of MET expression on outcome in BRAF(V600E/K) advanced melanoma.
Preclinical data suggest that signalling through the HGF-MET pathway may confer resistance to BRAF inhibition in BRAF(V600E/K) melanoma. Therefore, blockade of HGF-MET signalling might be a valid therapeutic strategy, in combination with BRAF inhibition, in BRAF(V600E/K) melanoma. The aim of this study was to investigate the clinical relevance of these observations by evaluating the survival impact of MET expression in patients with BRAF(V600E/K) advanced melanoma treated with vemurafenib. Formalin-fixed tissue blocks were obtained of tumours from patients enrolled in the BRIM2 (n = 59) and BRIM3 (n = 150) trials of vemurafenib in advanced BRAF(V600E/K) melanoma. Immunohistochemistry for MET (SP44 rabbit monoclonal antibody) was performed with a highly validated assay and clinically validated scoring system. Pretreatment MET expression was frequent at the ≥1 + cutoff (BRIM3, 31%; BRIM2, 49%), but relatively infrequent at the ≥2 + cutoff (BRIM3, 9%; BRIM2, 19%). Retrospective subset analyses showed that, irrespective of the cutoff used or the treatment arm, MET expression did not show prognostic significance, in terms of objective response rate, progression-free survival, or overall survival. MET is expressed in a proportion of BRAF(V600E/K) advanced melanomas. Further analyses on appropriately powered subsets are needed to determine the prognostic and predictive significance of MET in vemurafenib-treated melanoma.


24(S)-hydroxycholesterol induces neuronal cell death through necroptosis, a form of programmed necrosis.
24(S)-Hydroxycholesterol (24S-OHC) produced by cholesterol 24-hydroxylase expressed mainly in neurons plays an important physiological role in the brain. Conversely, it has been reported that 24S-OHC possesses potent cytotoxicity. The molecular mechanisms of 24S-OHC-induced cell death have not yet been fully elucidated. In this study, using human neuroblastoma SH-SY5Y cells and primary cortical neuronal cells derived from rat embryo, we characterized the form of cell death induced by 24S-OHC. SH-SY5Y cells treated with 24S-OHC exhibited neither fragmentation of the nucleus nor caspase activation, which are the typical characteristics of apoptosis. 24S-OHC-treated cells showed necrosis-like morphological changes but did not induce ATP depletion, one of the features of necrosis. When cells were treated with necrostatin-1, an inhibitor of receptor-interacting serine/threonine kinase 1 (RIPK1) required for necroptosis, 24S-OHC-induced cell death was significantly suppressed. The knockdown of RIPK1 by transfection of small interfering RNA of RIPK1 effectively attenuated 24S-OHC-induced cell death. It was found that neither SH-SY5Y cells nor primary cortical neuronal cells expressed caspase-8, which was regulated for RIPK1-dependent apoptosis. Collectively, these results suggest that 24S-OHC induces neuronal cell death by necroptosis, a form of programmed necrosis.


Formation of fe cluster superlattice in a metal-organic quantum-box network.
We report on the self-assembly of Fe adatoms on a Cu(111) surface that is patterned by a metal-organic honeycomb network, formed by coordination of dicarbonitrile pentaphenyl molecules with Cu adatoms. Fe atoms landing on the metal surface are mobile and steered by the quantum confinement of the surface state electrons towards the center of the network hexagonal cavities. In cavities hosting more than one Fe, preferential interatomic distances are observed. The adatoms in each hexagon aggregate into a single cluster upon gentle annealing. These clusters are again centered in the cavities and their size is discerned by their distinct apparent heights.


Octopaminergic system in the brain controls aggressive motivation in the ant, Formica japonica.
The ant, Formica japonica, is polyphagous and workers hunt other insects as foods. In this study, interspecific aggression was examined in the workers and queens. Behavior experiments demonstrated that interspecific aggressiveness was significantly higher in workers than queens. Workers showed predatory aggressive behavior towards crickets, on the other hand, queens elicited threat behavior but they didn't attack crickets. In order to investigate neuronal mechanisms underlying regulation of aggressive motivation, the role of biogenic amine in the brain in evoking aggressive behavior was examined by measuring biogenic amine using high-performance liquid chromatography (HPLC) with electrochemical detection (ECD). No significant difference in the octopamine (OA) level was found between workers and queens, but the level of N-acetyloctopamine (NacOA) in the brain of queens was significantly higher than that of workers. This study suggests that OAergic system in the brain must involve in controlling aggressive motivation in the ants.


Odd men out: a quantitative objective procedure for identifying anomalous members of a set of noisy images of ostensibly identical specimens.
Achievement of an optimal improvement in signal-to-noise ratio from image averaging techniques depends crucially on the assumption that all members of the set of images to be averaged are fundamentally alike. In HREM of biological macromolecules, this assumption may be invalid for such reasons as variations in viewing geometry, non-uniformity of staining, or structural perturbations caused by specimen preparation procedures of radiation damage. Inclusion of data that are compromized these or other factors will degrade the information content of the averaged image. Here we present an algorithm which provides an objective quantitative method for the identification and elimination of anomalous members of a set of pre-aligned images. Based on a statistical criterion of mutual consistency, the algorithm forms an ordered list in which the individual images are ranked from most to least reliable. On specification of the noise statistics--in the formulation given here, of stationary white noise--an acceptability threshold in this ordered list is imposed. The derivation and implementation of this algorithm are presented, its properties discussed, and its application illustrated using both real and model electron micrograph data.


Macrophage polykaryon formation in vitro by peritoneal cells from mice given injections of sodium periodate.
Peritoneal macrophages from mice that have received two separate intraperitoneal injections of the sterile, soluble oxidant NaIO4 form macrophage polykaryons (MPs) in vitro, but peritoneal macrophage from untreated, peptone-treated, or mice infected with bacille Calmette-Guerin (BCG) do not. The polykaryons are noted after 18-24 hours of culture and continue to form over a 60-72-hour period. The MPs do not form if the macrophage density is less than 4 x 10(3)/sq mm. The polykaryons appear in vitro only in cultures with less than or equal to 1-5 ng/ml lipopolysaccharide (LPS) (amounts of LPS that commonly contaminate culture medium and serum). Hydrocortisone hemisuccinate (2.6 x 10(-9) M) inhibits MP formation in vitro. Lymphocytes do not influence the polykaryon formation, and supernatants from MP cultures do not cause fusion of other macrophages. Microcinephotography demonstrates fusion of the macrophages to form the large polykaryons, which are less motile than uninuclear macrophages. The polykaryons assume different forms; generally, the nuclei (mean, 16.8 nuclei/MP; range, 2-137 nuclei/MP) are centrally located, and the nuclear chromatin of all nuclei appears similar. The MPs phagocytize polystyrene spheres and glutaraldehyde-treated erythrocytes to the same degree as do uninuclear macrophages when determined as particles per nucleus (phagocytic index), but their phagocytic index of IgG-coated erythrocytes is decreased. Peritoneal macrophages from mice given double injections of NaIO4 are nontumoricidal in the absence of LPS, but LPS, in amounts sufficient to inhibit polykaryon formation, renders the macrophages tumoricidal. Populations of macrophages containing MPs formed over 3 days of cultures also respond to LPS or macrophage activating factor (MAF) to demonstrate enhanced tumoricidal activity.


Evaluation of infectious titer in a candidate HSV type 2 vaccine by a quantitative molecular approach.
One of the critical tasks in analytical testing is to monitor and assign the infectivity or potency of viral based vaccines from process development to production of final clinical lots. In this study, a high throughput RT-qPCR based approach was developed to evaluate the infectious titre in a replication-defective HSV-2 candidate vaccine, called HSV529. This assay is a combination of viral propagation and quantitative RT-PCR which measures the amount of RNA in infected cells after incubation with test samples. The relative infectious titre of HSV529 candidate vaccine was determined by a RT-qPCR method targeting HSV-2 gD2 gene. The data were analyzed using the parallel-line analysis as described in the European Pharmacopoeia 8th edition. The stability of HSV529 test samples were also investigated in a concordance study between RT-qPCR infectivity assay and a classical plaque assays. A suitable correlation was determined between both assays using an identical sample set in both assays. The RT-qPCR infectivity assay was further characterized by evaluating the intermediate precision and accuracy. The coefficient of variation from the six independent assays was less than 10%. The accuracy of each of the assay was also evaluated in the range of 92.91% to 120.57%. Our data demonstrate that the developed RT-qPCR infectivity assay is a rapid high throughput approach to quantify the infectious titer or potency of live attenuated or defective viral-based vaccines, an attribute which is associated with product quality.


Spinal cord monitoring in scoliosis surgery. Experience with 1168 cases.
Since 1981, during operations for spinal deformity, we have routinely used electrophysiological monitoring of the spinal cord by the epidural measurement of somatosensory evoked potentials (SEPs) in response to stimulation of the posterior tibial nerve. We present the results in 1168 consecutive cases. Decreases in SEP amplitude of more than 50% occurred in 119 patients, of whom 32 had clinically detectable neurological changes postoperatively. In 35 cases the SEP amplitude was rapidly restored, either spontaneously or by repositioning of the recording electrode; they had no postoperative neurological changes. One patient had delayed onset of postoperative symptoms referrable to nerve root lesions without evidence of spinal cord involvement, but there were no false negative cases of intra-operative spinal cord damage. In 52 patients persistent, significant, SEP changes were noted without clinically detectable neurological sequelae. None of the many cases which showed falls in SEP amplitude of less than 50% experienced neurological problems. Neuromuscular scoliosis, the use of sublaminar wires, the magnitude of SEP decrement, and a limited or absent intra-operative recovery of SEP amplitude were identified as factors which increased the risk of postoperative neurological deficit.


Consequences of phloem pathway unloading/reloading on equilibrium flows between source and sink: a modelling approach.
It is now accepted that the transport phloem, linking major sources and sinks, is leaky, and this leakage can be considerable. Hence for phloem transport to function over the long distances observed, a large fraction of this unloaded photosynthate must be reloaded. A fraction of this unloaded solute is used to maintain tissues surrounding the phloem, as well as being stored. Also, pathway unloading/reloading acts as a short-term buffer to source and sink changes. In this work we present the first attempt to include both pathway unloading and reloading of carbohydrate in the modelling of pressure driven flow to determine if this has any significant effect upon source-sink dynamics. Our results indicated that the flow does not follow Poiseuille dynamics, and that pathway unloading alters the solute concentration and hydrostatic pressure profiles. Hence, measurement of either of these without considerable other detail tells us very little about the flow mechanisms. With adequate reloading along the pathway, the effects of pathway unloading can completely compensate for, making the entire system look like one with no pathway unloading.


Visuospatial processing in memory for word location in writing.
Two experiments examined how visuospatial processing engaged during text composition intervenes in memory for word location. Experiment 1 showed that in contrast to participants who performed a spatial task concurrently with composing a text, participants who performed a concurrent visual task recalled fewer word locations after the composition. Consequently, it is hypothesized that writers process the written text in order to visually represent its physical layout, and that this representation is then used when locating words. Experiment 2 tested this hypothesis by comparing a standard composition condition (with the written trace) with a condition in which the written trace was suppressed during composition, and with a condition without written trace and with added visual noise. Memory for word location only decreased with visual noise, indicating that construction of the visual representation of the text does not rely on the written trace but involves visual working memory.


All major cholesterol-dependent cytolysins use glycans as cellular receptors.
Cholesterol-dependent cytolysins (CDCs) form pores in cholesterol-rich membranes, but cholesterol alone is insufficient to explain their cell and host tropism. Here, we show that all eight major CDCs have high-affinity lectin activity that identifies glycans as candidate cellular receptors. Streptolysin O, vaginolysin, and perfringolysin O bind multiple glycans, while pneumolysin, lectinolysin, and listeriolysin O recognize a single glycan class. Addition of exogenous carbohydrate receptors for each CDC inhibits toxin activity. We present a structure for suilysin domain 4 in complex with two distinct glycan receptors, P1 antigen and αGal/Galili. We report a wide range of binding affinities for cholesterol and for the cholesterol analog pregnenolone sulfate and show that CDCs bind glycans and cholesterol independently. Intermedilysin binds to the sialyl-TF O-glycan on its erythrocyte receptor, CD59. Removing sialyl-TF from CD59 reduces intermedilysin binding. Glycan-lectin interactions underpin the cellular tropism of CDCs and provide molecular targets to block their cytotoxic activity.


Co-Infection with Common Respiratory Pathogens and SARS-CoV-2 in Patients with COVID-19 Pneumonia and Laboratory Biochemistry Findings: A Retrospective Cross-Sectional Study of 78 Patients from a Single Center in China.
BACKGROUND This retrospective study aimed to investigate co-infections with common respiratory pathogens and SARS-CoV-2 and laboratory biochemistry findings in patients with COVID-19 in the Zhuzhou area of China, in order to provide a reference for the disease assessment and clinical treatment of COVID-19. MATERIAL AND METHODS The clinical data of COVID-19 patients admitted to the hospital of Zhuzhou City from January 28 to March 15, 2020, as well as laboratory test results for respiratory pathogens and biochemical indicators, were collected to conduct correlation analyses. All patients were diagnosed based on fluorescence-based PCR assay for SARS-CoV-2. RESULTS Eleven of the 78 patients (14.1%) were co-infected with other respiratory pathogens, among which Mycoplasma pneumoniae (n=5, 45.5%) and respiratory syncytial virus (n=4, 36.4%) were the most frequent. There were 8 patients co-infected with 1 other pathogen and 3 patients co-infected with 2 other pathogens. Compared with mono-infected COVID-19 patients, patients with co-infections had significantly higher levels of procalcitonin (P=0.002). CONCLUSIONS The findings showed that Mycoplasma pneumonia and respiratory syncytial virus were the most common co-infections in patients with COVID-19 pneumonia. Increased levels of PCT in patients with COVID-19 pneumonia were associated with co-infection.


Anti-cooperative assembly of the SRP19 and SRP68/72 components of the signal recognition particle.
The mammalian SRP (signal recognition particle) represents an important model for the assembly and role of inter-domain interactions in complex RNPs (ribonucleoproteins). In the present study we analysed the interdependent interactions between the SRP19, SRP68 and SRP72 proteins and the SRP RNA. SRP72 binds the SRP RNA largely via non-specific electrostatic interactions and enhances the affinity of SRP68 for the RNA. SRP19 and SRP68 both bind directly and specifically to the same two RNA helices, but on opposite faces and at opposite ends. SRP19 binds at the apices of helices 6 and 8, whereas the SRP68/72 heterodimer binds at the three-way junction involving RNA helices 5, 6 and 8. Even though both SRP19 and SRP68/72 stabilize a similar parallel orientation for RNA helices 6 and 8, these two proteins bind to the RNA with moderate anti-cooperativity. Long-range anti-cooperative binding by SRP19 and SRP68/72 appears to arise from stabilization of distinct conformations in the stiff intervening RNA scaffold. Assembly of large RNPs is generally thought to involve either co-operative or energetically neutral interactions among components. By contrast, our findings emphasize that antagonistic interactions can play significant roles in assembly of multi-subunit RNPs.


Targeted metabolomics reveals the association between central carbon metabolism and pulmonary nodules.
With the widespread application of low-dose computed tomography (LDCT) technology, pulmonary nodules have aroused more attention. Significant alteration in plasma metabolite levels, mainly amino acid and lipid, have been observed in patients of PNs. However, evidence on the association between central carbon metabolism and PNs are largely unknown. The aim of this study was to investigate the underlying association of PNs and plasma central carbon metabolites. We measured the levels of 16 plasma central carbon metabolites in 1954 participants who gained LDCT screening in MALSC cohort. The inverse probability weighting (IPW) technique was used to control for bias due to self-selection for LDCT in the assessed high-risk population. The least absolute shrinkage and selection operator (LASSO) penalized regression was used to deal with the problem of multicollinearity among metabolites and the combined association of central carbon metabolites with PNs was estimated by using quantile g-computation (QgC) models. A quartile increase in 3-hydroxybutyric acid, gluconic acid, succinic acid and hippuric acid was positively associated with the PNs risk, whereas a quartile increase in 2-oxadipic acid and fumaric acid was negatively associated with the risk of PNs in multiple-metabolite models. A positive but insignificant joint associations of the mixture of 16 metabolites with PNs was observed by using QgC models analyses. Further studies are warranted to clarify the association between circulating metabolites and PNs and the biological mechanisms.


Calcium signals recorded from two new purpurate indicators inside frog cut twitch fibers.
Two new Ca indicators, purpurate-3,3'diacetic acid (PDAA) and 1,1'-dimethylpurpurate-3,3'diacetic acid (DMPDAA), were synthesized and used to measure Ca transients in frog cut muscle fibers. These indicators are analogues of the purpurate components of murexide and tetramethylmurexide, in which two acetate groups have been incorporated into each molecule to render it membrane impermeant. The apparent dissociation constant for Ca is 0.95 mM for PDAA and 0.78 mM for DMPDAA. One of the indicators was introduced into a cut fiber, which was mounted in a double Vaseline-gap chamber, by diffusion from the end-pool solutions. The time course of indicator concentration, monitored optically in the middle of the fiber in the central-pool region, suggests that 19% of the PDAA or 27% of the DMPDAA became bound or sequestered inside the fiber. In resting fibers, the absorbance spectrum of either indicator was well fitted by the indicator's [Ca] = 0 mM cuvette absorbance spectrum, which is consistent with the idea that PDAA and DMPDAA do not enter the sarcoplasmic reticulum as tetramethylmurexide appears to be able to do (Maylie, J., M. Irving, N.L. Sizto, G. Boyarsky, and W. K. Chandler, 1987. Journal of General Physiology. 89:145-176). After an action potential, the absorbance of either indicator underwent a rapid and transient change that returned to the prestimulus baseline within 100-200 ms. The amplitude of this change had a wavelength dependence that matched the indicator's Ca-difference spectrum. The average amplitude of peak free [Ca] was 21 microM (PDAA or DMPDAA) if all the indicator inside a fiber was able to react with Ca as in cuvette calibrations, and was 26 (PDAA) or 28 microM (DMPDAA) if only freely diffusible indicator could so react. These results suggest that PDAA and DMPDAA are the first Ca indicators that provide a reliable estimate of both the amplitude and time course of (the spatial average of) free [Ca] in a twitch muscle fiber after an action potential.


Baseball outfielders maintain a linear optical trajectory when tracking uncatchable fly balls.
The authors investigated whether behavior of fielders pursuing uncatchable fly balls supported either (a) maintenance of a linear optical trajectory (LOT) with monotonic increases in optical ball height or (b) maintenance of optical acceleration cancellation (OAC) with simultaneous lateral alignment with the ball. Past work supports usage of both LOT and OAC strategies in the pursuit of catchable balls headed to the side. When balls are uncatchable, fielders must choose either optical linearity or alignment at the expense of the other. Fielders maintained the LOT strategy more often and for a longer period of time than they did the OAC alignment strategy. Findings support the LOT strategy as primary when pursuing balls headed to the side, whether catchable or not.


Phosphodiesterase inhibitors say NO to Alzheimer's disease.
Phosphodiesterases (PDEs) consisted of 11 subtypes (PDE1 to PDE11) and over 40 isoforms that regulate levels of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), the second messengers in cell functions. PDE inhibitors (PDEIs) have been attractive therapeutic targets due to their involvement in diverse medical conditions, e.g. cardiovascular diseases, autoimmune diseases, Alzheimer's disease (AD), etc. Among them; AD with a complex pathology is a progressive neurodegenerative disorder which affect mostly senile people in the world and only symptomatic treatment particularly using cholinesterase inhibitors in clinic is available at the moment for AD. Consequently, novel treatment strategies towards AD are still searched extensively. Since PDEs are broadly expressed in the brain, PDEIs are considered to modulate neurodegenerative conditions through regulating cAMP and cGMP in the brain. In this sense, several synthetic or natural molecules inhibiting various PDE subtypes such as rolipram and roflumilast (PDE4 inhibitors), vinpocetine (PDE1 inhibitor), cilostazol and milrinone (PDE3 inhibitors), sildenafil and tadalafil (PDE5 inhibitors), etc have been reported showing encouraging results for the treatment of AD. In this review, PDE superfamily will be scrutinized from the view point of structural features, isoforms, functions and pharmacology particularly attributed to PDEs as target for AD therapy.


[Functional properties and cytoskeletal-dependent regulation of sodium channels in leukemia cell membranes].
The paper is devoted to membrane mechanisms of sodium influx from the extracellular medium to the cytoplasm in nonexcitable cells. With the use of patch clamp technique, the activity of non-voltage-gated ionic channels in plasma membrane of human leukemia K562 cells was examined. We have identified two types of Na-permeable channels characterized by unitary conductance of 12 pS and differing in their selectivity among monovalent cations. A relative permeability value PNa/PK was estimated for both types referred to as channels of high (HS, PNa/PK = 10) and low (LS, PNa/PK = 3) selectivity, resp. Both the channels were impermeable to bivalent cations (Ca2+, Ba2+), not blocked by tetradotoxin. Their sensitivity to amiloride was extremely low. Cytochalasin D treatment of cells resulted in a significant increase in the activity of LS Na-conducting channels. Application of exogenous gelsolin to the cytoplasmic surface of inside-out membrane patch at free Ca2+ level of 1 mkM induced a similar effect of sodium channel activation; the subsequent addition of actin reduced the channel activity up to the background level. Our results show that the cortical F-actin network plays an important role in regulating the novel family of sodium channels in nonexcitable cells. It could be assumed that the actin disassembly causes a rise in LS channel activity, whereas the actin assembly induces inactivation of the channels.


[Influence of pilot's professional activity on the adaptive character of hemodynamic reactions to the tilt test].
The comparative study of blood circulation reaction to the tilt test was performed with three groups of essentially healthy subjects: AF pilots (n = 72), locomen (n = 44) and students of Military Institute of Radio Electronics (n = 32). In each group, adequate adaptive reactions were due to the eukinetic hemodynamics with steady minute volume and blood pressure values combined with heart rate increase by 10-12%. Hypokinetic hemodynamics caused 20% growth in minute volume during the initial 5 minutes of tilting in pilots versus 8-12% growth in non-flyers. Hyperkinetic hemodynamics was responsible for the minute volume growth in pilots but not in the other groups of subjects. Parasympathetic system activation was peculiar of hypokinetic hemodynamics, while sympathetic system activation was associated with hyperkinetic hemodynamics. These results point to a larger body functional reserve in pilots compared with the other subjects as a by-effect of the profession.


Towards conservation of omani local chicken: phenotypic characteristics, management practices and performance traits.
Characterizing local chicken types and their mostly rural production systems is prerequisite for designing and implementing development and conservation programs. This study evaluated the management practices of small-scale chicken keepers and the phenotypic and production traits of their chickens in Oman, where conservation programs for local livestock breeds have currently started. Free-range scavenging was the dominant production system, and logistic regression analysis showed that socio-economic factors such as training in poultry keeping, household income, income from farming and gender of chicken owners influenced feeding, housing, and health care practices (p<0.05). A large variation in plumage and shank colors, comb types and other phenotypic traits within and between Omani chicken populations were observed. Male and female body weight differed (p<0.05), being 1.3±0.65 kg and 1.1±0.86 kg respectively. Flock size averaged 22±7.7 birds per household with 4.8 hens per cock. Clutch size was 12.3±2.85 and annual production 64.5±2.85 eggs per hen. Egg hatchability averaged 88±6.0% and annual chicken mortality across all age and sex categories was 16±1.4%. The strong involvement of women in chicken keeping makes them key stakeholders in future development and conservation programs, but the latter should be preceded by a comprehensive study of the genetic diversity of the Omani chicken populations.


Exclusion and control: patterns aimed at limiting the influence of faculty of color.
This article reports on findings from a grounded theory study of the experiences of faculty of color (FOC) in predominantly Euro-American schools of nursing. Findings indicate that a dominant group sometimes referred to as the "Good Old Girls" posed a major barrier to change. To maintain the status-quo, the Good Old Girls used their influence to subject FOC to Patterns of Exclusion and Control with the goal of controlling the influence of FOC. This occurred through parallel and interrelated exclusion and control subprocesses. Exclusion and control strategies threatened FOC's success in academe while simultaneously jeopardizing their well-being. The exclusion strategies FOC experienced included Invalidation of Sense of Self, Othering, and Unequal Standards and Access to Resources. The control strategies included Insincerity and Putting You in Your Place. We describe each of these processes in detail and end with a discussion of the implications of our findings for educational practice.


Directed Evolution of Canonical Loops and Their Swapping between Unrelated Serine Proteinase Inhibitors Disprove the Interscaffolding Additivity Model.
Reversible serine proteinase inhibitors comprise 18 unrelated families. Each family has a distinct representative structure but contains a surface loop that adopts the same, canonical conformation in the enzyme-inhibitor complex. The Laskowski mechanism universally applies for the action of all canonical inhibitors independent of their scaffold, but it has two nontrivial extrapolations. Intrascaffolding additivity states that all enzyme-contacting loop residues act independently of each other, while interscaffolding additivity claims that these residues act independently of the scaffold. These theories have great importance for engineering proteinase inhibitors but have not been comprehensively challenged. Therefore, we tested the interscaffolding additivity theory by hard-randomizing all enzyme-contacting canonical loop positions of a Kazal- and a Pacifastin-scaffold inhibitor, displaying the variants on M13 phage, and selecting the libraries on trypsin and chymotrypsin. Directed evolution delivered different patterns on both scaffolds against both enzymes, which contradicts interscaffolding additivity. To quantitatively assess the extent of non-additivity, we measured the affinities of the optimal binding loop variants and their binding loop-swapped versions. While optimal variants have picomolar affinities, swapping the evolved loops results in up to 200,000-fold affinity loss. To decipher the underlying causes, we characterized the stability, overall structure and dynamics of the inhibitors with differential scanning calorimetry, circular dichroism and NMR spectroscopy and molecular dynamic simulations. These studies revealed that the foreign loop destabilizes the lower-stability Pacifastin scaffold, while the higher-stability Kazal scaffold distorts the foreign loop. Our findings disprove interscaffolding additivity and show that loop and scaffold form one integrated unit that needs to be coevolved to provide high-affinity inhibition.


Application of bacteriophages to reduce Salmonella contamination on workers' boots in rendering-processing environment.
Workers' boots are considered one of the re-contamination routes of Salmonella for rendered meals in the rendering-processing environment. This study was conducted to evaluate the efficacy of a bacteriophage cocktail for reducing Salmonella on workers' boots and ultimately for preventing Salmonella re-contamination of rendered meals. Under laboratory conditions, biofilms of Salmonella Typhimurium avirulent strain 8243 formed on rubber templates or boots were treated with a bacteriophage cocktail of 6 strains (ca. 9 log PFU/mL) for 6 h at room temperature. Bacteriophage treatments combined with sodium hypochlorite (400 ppm) or 30-second brush scrubbing also were investigated for a synergistic effect on reducing Salmonella biofilms. Sodium magnesium (SM) buffer and sodium hypochlorite (400 ppm) were used as controls. To reduce indigenous Salmonella on workers' boots, a field study was conducted to apply a bacteriophage cocktail and other combined treatments 3 times within one wk in a rendering-processing environment. Prior to and after bacteriophage treatments, Salmonella populations on the soles of rubber boots were swabbed and enumerated on XLT-4, Miller-Mallinson or CHROMagar™ plates. Under laboratory conditions, Salmonella biofilms formed on rubber templates and boots were reduced by 95.1 to 99.999% and 91.5 to 99.2%, respectively. In a rendering-processing environment (ave. temperature: 19.3°C; ave. relative humidity: 48%), indigenous Salmonella populations on workers' boots were reduced by 84.2, 92.9, and 93.2% after being treated with bacteriophages alone, bacteriophages + sodium hypochlorite, and bacteriophages + scrubbing for one wk, respectively. Our results demonstrated the effectiveness of bacteriophage treatments in reducing Salmonella contamination on the boots in both laboratory and the rendering-processing environment.


[Development and evaluation of a serological protocol of fluorescence polarization for the preliminary study of Brucella spp antibodies in humans].
In order to show the development and scope of a serological analysis method based on fluorescence polarization assay (FPA) from a drop of blood obtained by the capillary technique, a Brucella antibody assay was performed on a group of 321 high-risk workers. The results were compared with data from the analysis of blood serum by FPA and a competitive enzyme immunoassay (ELISA-c). The number of concordance was 318 (99.06%), and discordant 3 (0.93%), which were negative in serum by fluorescence polarization (FPAs) and ELISA-c, but positive with capillary FPA (FPAc). The comparative results FPAc were: sensitivity 100%; specificity: 99.05%; positive predictive value 66.67%; negative predictive value 100.0%; false positive rate: 0.95%; false negative rate: 0%; accuracy: 98.0%; odds ratio: 203.00. The youden J for both FPA methods was 0.667. The identification was considered reliable and the correlation of both procedures, FPA and ELISA-c, was no statistically different (P > 0.05%), which allows to highly recommend the study implementation of human brucellosis with capillary blood as a preliminary method.


[Acute meningoencephalitis in children caused by herpes simplex virus type I].
In 14 observations of acute meningoencephalitis in children mainly under 3 years of the age, data pointing to an etiological role of herpes simplex type I virus, were obtained on laboratory examination. Six children died, and in the rest 8 children who survived gross residua were observed. The morphological picture showed extensive colliquation necroses in the parietal, temporal, and less frequently, in occipital lobes and the Varolian pons. In 4 cases out of 6 intranuclear inclusions were detected. The incidence of herpetic encephalitis was 16%.


GmPIN1-mediated auxin asymmetry regulates leaf petiole angle and plant architecture in soybean.
Crop breeding during the Green Revolution resulted in high yields largely due to the creation of plants with semi-dwarf architectures that could tolerate high-density planting. Although semi-dwarf varieties have been developed in rice, wheat and maize, none was reported in soybean (Glycine max), and few genes controlling plant architecture have been characterized in soybean. Here, we demonstrate that the auxin efflux transporter PINFORMED1 (GmPIN1), which determines polar auxin transport, regulates the leaf petiole angle in soybean. CRISPR-Cas9-induced Gmpin1abc and Gmpin1bc multiple mutants displayed a compact architecture with a smaller petiole angle than wild-type plants. GmPIN1 transcripts and auxin were distributed asymmetrically in the petiole base, with high levels of GmPIN1a/c transcript and auxin in the lower cells, which resulted in asymmetric cell expansion. By contrast, the (iso)flavonoid content was greater in the upper petiole cells than in the lower cells. Our results suggest that (iso)flavonoids inhibit GmPIN1a/c expression to regulate the petiole angle. Overall, our study demonstrates that a signal cascade that integrates (iso)flavonoid biosynthesis, GmPIN1a/c expression, auxin accumulation, and cell expansion in an asymmetric manner creates a desirable petiole curvature in soybean. This study provides a genetic resource for improving soybean plant architecture.


The Use of Interdisciplinary Approaches to Understand the Biology of Campylobacter jejuni.
Campylobacter jejuni is a bacterial pathogen recognised as a major cause of foodborne illness worldwide. While Campylobacter jejuni generally does not grow outside its host, it can survive outside of the host long enough to pose a health concern. This review presents an up-to-date description and evaluation of biological, mathematical, and statistical approaches used to understand the behaviour of this foodborne pathogen and suggests future avenues which can be explored. Specifically, the incorporation of mathematical modelling may aid the understanding of C. jejuni biofilm formation both outside and inside the host. Predictive studies may be improved by the introduction of more standardised protocols for assessments of disinfection methods and by assessment of novel physical disinfection strategies as well as assessment of the efficiency of plant extracts on C. jejuni eradication. A full description of the metabolic pathways of C. jejuni, which is needed for the successful application of metabolic models, is yet to be achieved. Finally, a shift from animal models (except for those that are a source of human campylobacteriosis) to human-specific data may be made possible due to recent technological advancements, and this may lead to more accurate predictions of human infections.


Mothers with serious mental illness: their experience of "hitting bottom".
This study sought to understand the experience of "hitting bottom" from the perspective of 32 mothers with serious mental illness. Secondary narrative analysis of 173 stories about experiences related to hitting bottom were identified. Enactment of their perceived mothering roles and responsibilities was compromised when confronted by the worst of illness. Subsequent to women's descent to bottom was their need for a timely and safe exit from bottom. An intense experience in bottom further jeopardized their parenting and treatment self-determination and, for some, their potential for survival. The results suggest that prevention of bottom is feasible with early assessment of the diverse issues contributing to mothers' vulnerabilities. Interventions to lessen their pain may circumvent bottom experiences. Healing necessitates purposeful approaches to minimize the private and public trauma of bottom experiences, nurture growth towards a future, and establish resources to actualize such a life.


From sensors to spikes: evolving receptive fields to enhance sensorimotor information in a robot-arm.
In biological systems, instead of actual encoders at different joints, proprioception signals are acquired through distributed receptive fields. In robotics, a single and accurate sensor output per link (encoder) is commonly used to track the position and the velocity. Interfacing bio-inspired control systems with spiking neural networks emulating the cerebellum with conventional robots is not a straight forward task. Therefore, it is necessary to adapt this one-dimensional measure (encoder output) into a multidimensional space (inputs for a spiking neural network) to connect, for instance, the spiking cerebellar architecture; i.e. a translation from an analog space into a distributed population coding in terms of spikes. This paper analyzes how evolved receptive fields (optimized towards information transmission) can efficiently generate a sensorimotor representation that facilitates its discrimination from other "sensorimotor states". This can be seen as an abstraction of the Cuneate Nucleus (CN) functionality in a robot-arm scenario. We model the CN as a spiking neuron population coding in time according to the response of mechanoreceptors during a multi-joint movement in a robot joint space. An encoding scheme that takes into account the relative spiking time of the signals propagating from peripheral nerve fibers to second-order somatosensory neurons is proposed. Due to the enormous number of possible encodings, we have applied an evolutionary algorithm to evolve the sensory receptive field representation from random to optimized encoding. Following the nature-inspired analogy, evolved configurations have shown to outperform simple hand-tuned configurations and other homogenized configurations based on the solution provided by the optimization engine (evolutionary algorithm). We have used artificial evolutionary engines as the optimization tool to circumvent nonlinearity responses in receptive fields.


Current levels and composition profiles of emerging halogenated flame retardants and dehalogenated products in sewage sludge from municipal wastewater treatment plants in China.
Occurrence of new toxic chemicals in sludge from wastewater treatment plants (WWTPs) is of concern for the environment and human health. Alternative halogenated flame retardants (HFRs) are a group of potentially harmful organic contaminants in the environment. In this study, a nationwide survey was carried out to identify the occurrence of HFRs and their potential dehalogenated products in sewage sludge from 62 WWTPs in China. Of all 20 target chemicals analyzed, decabromodiphenyl ethane (DBDPE), hexabromocyclododecane (HBCD) and 1, 2-bis (2,4,6-tribromophenoxy)-ethane (BTBPE) were detected in all sludge samples, and the concentrations were in the range of 0.82-215, 0.09-65.8, and 0.10-2.26 ng g(-1) d.w., respectively. Dechlorane Plus (DP) was found in 60 of 62 samples, and the concentration ranged from nd-298 ng g(-1) with a mean of 18.9 ng g(-1) d.w. The anti-DP fractional abundance fanti (0.79) in the samples was much higher than the commercial DP composition (fanti=0.59), indicating a stereoselective degradation. Comparison with global sludge concentrations of HFRs indicate that China is at the medium pollution level in the world. Principal components analysis revealed that strong correlations existed between ln-transformed concentrations (natural logarithm) of the dominant BFRs and total organic carbon (TOC) as well as industrial wastewater proportion, influent volume and serving population. Significant linear relationships (R=0.360-0.893, p<0.01) were found among emerging brominated flame retardants (BFRs), suggesting their common commercial applications and release sources to the environment. Two kinds of dehalogenated products, pentabromocyclododecane (PBCD) and undecachloropentacyclooctadecadiene (Cl11-DP), derived from HBCD and DP, were also identified in sewage sludge for the first time.


Reduced albuminuria with sarpogrelate is accompanied by a decrease in monocyte chemoattractant protein-1 levels in type 2 diabetes.
Sarpogrelate has been shown to reduce albuminuria in diabetic nephropathy. For examination of whether this is based on the same mechanisms as angiotensin II receptor blockers or thiazolidinedione, effects of sarpogrelate on atherosclerotic inflammatory molecules and their relations to albuminuria in patients who had diabetes and had already been treated with angiotensin II receptor blockers and with or without thiazolidinedione were examined. Forty patients who had diabetes with nephropathy and arteriosclerosis obliterans and had already been treated with angiotensin II receptor blocker (n = 40) were randomly assigned to sarpogrelate (300 mg/d; n = 20) or aspirin group (100 mg/d; n = 20). Plasma monocyte chemoattractant protein-1 and urinary albumin-to-creatinine ratio and monocyte chemoattractant protein-1 were measured at baseline and 16 wk after administration. Only the sarpogrelate group showed increases in plasma adiponectin and decreases in both plasma and urinary monocyte chemoattractant protein-1 and albumin-to-creatinine ratio levels. Moreover, percentage change of monocyte chemoattractant protein-1 level correlated positively to that of albumin-to-creatinine ratio. Even when the sarpogrelate group was further divided into two groups with (n = 9) or without thiazolidinedione (n = 11), changes in monocyte chemoattractant protein-1 or albumin-to-creatinine ratio did not differ. Sarpogrelate can reduce albuminuria and plasma and urinary monocyte chemoattractant protein-1 levels while increasing plasma adiponectin in diabetic nephropathy. These effects seem to be mediated via mechanisms that are different from those of angiotensin II receptor blocker or thiazolidinedione.


Complete Genome Sequences of Sequence Type 71 (ST71) and ST97 Staphylococcus aureus Isolates from Bovine Milk.
This is the announcement of draft genome sequences for Staphylococcus aureus strains belonging to sequence type 97 (ST97) and ST71. These sequence types are commonly associated with bovine mastitis, and the strains were isolated in Ireland in 2010 from the milk of cows with clinical mastitis.


Identification of destabilizing and stabilizing mutations of Ste2p, a G protein-coupled receptor in Saccharomyces cerevisiae.
The isolation of mutations affecting the stabilities of transmembrane proteins is useful for enhancing the suitability of proteins for structural characterization and identification of determinants of membrane protein stability. We have pursued a strategy for the identification of stabilized variants of the yeast α-factor receptor Ste2p. Because it was not possible to screen directly for mutations providing thermal stabilization, we first isolated a battery of destabilized temperature-sensitive variants, based on loss of signaling function and decreased levels of binding of the fluorescent ligand, and then screened for intragenic second-site suppressors of these phenotypes. The initial screens recovered singly and multiply substituted mutations conferring temperature sensitivity throughout the predicted transmembrane helices of the receptor. All of the singly substituted variants exhibit decreases in cell-surface expression. We then screened randomly mutagenized libraries of clones expressing temperature-sensitive variants for second-site suppressors that restore elevated levels of binding sites for fluorescent ligand. To determine whether any of these were global suppressors, and thus likely stabilizing mutations, they were combined with different temperature-sensitive mutations. Eight of the suppressors exhibited the ability to reverse the defect in ligand binding of multiple temperature-sensitive mutations. Combining certain suppressors into a single allele resulted in levels of suppression greater than that seen with either suppressor alone. Solubilized receptors containing suppressor mutations in the absence of temperature-sensitive mutations exhibit a reduced tendency to aggregate during immobilization on an affinity matrix. Several of the suppressors also exhibit allele-specific behavior indicative of specific intramolecular interactions in the receptor.


Job task characteristics of Australian emergency services volunteers during search and rescue operations.
Search and rescue operations are necessary in locating, assisting and recovering individuals lost or in distress. In Australia, land-based search and rescue roles require a range of physically demanding tasks undertaken in dynamic and challenging environments. The aim of the current research was to identify and characterise the physically demanding tasks inherent to search and rescue operation personnel within Australia. These aims were met through a subjective job task analysis approach. In total, 11 criterion tasks were identified by personnel. These tasks were the most physically demanding, frequently occurring and operationally important tasks to these specialist roles. Muscular strength was the dominant fitness component for 7 of the 11 tasks. In addition to the discrete criterion tasks, an operational scenario was established. With the tasks and operational scenario identified, objective task analysis procedures can be undertaken so that practitioners can implement evidence-based strategies, such as physical selection procedures and task-based physical training programs, commensurate with the physical demands of search and rescue job roles. Practitioner Summary: The identification of physically demanding tasks amongst specialist emergency service roles predicates health and safety strategies which can be incorporated into organisations. Knowledge of physical task parameters allows employers to mitigate injury risk through the implementation of strategies modelled on the precise physical demands of the role.


Population toxicokinetics of tetrachloroethylene.
In assessing the distribution and metabolism of toxic compounds in the body, measurements are not always feasible for ethical or technical reasons. Computer modeling offers a reasonable alternative, but the variability and complexity of biological systems pose unique challenges in model building and adjustment. Recent tools from population pharmacokinetics, Bayesian statistical inference, and physiological modeling can be brought together to solve these problems. As an example, we modeled the distribution and metabolism of tetrachloroethylene (PERC) in humans. We derive statistical distributions for the parameters of a physiological model of PERC, on the basis of data from Monster et al. (1979). The model adequately fits both prior physiological information and experimental data. An estimate of the relationship between PERC exposure and fraction metabolized is obtained. Our median population estimate for the fraction of inhaled tetrachloroethylene that is metabolized, at exposure levels exceeding current occupational standards, is 1.5% [95% confidence interval (0.52%, 4.1%)]. At levels approaching ambient inhalation exposure (0.001 ppm), the median estimate of the fraction metabolized is much higher, at 36% [95% confidence interval (15%, 58%)]. This disproportionality should be taken into account when deriving safe exposure limits for tetrachloroethylene and deserves to be verified by further experiments.


Degradation of atrial natriuretic factor in the rat.
The biologically active circulating form of atrial natriuretic factor (ANF) in the rat is the 28-amino-acid peptide ANF-(Ser-99-Tyr-126). Degradation of this peptide in vivo as well as in vitro, in whole blood, in plasma and by the isolated mesenteric artery was investigated. Studies in vivo in the rat demonstrated that the elimination and degradation of ANF was extremely fast: within 3 min more than 95% of the injected immunoreactive material was eliminated from circulation. The production of a short C-terminal peptide was detected on injection of 125I-ANF-(Ser-99-Tyr-126) into the rat. This peptide increased proportionately with incubation time. Experiments in vitro in the presence of whole blood or plasma did not cause any major destruction of ANF even after incubation for 60 min. After this prolonged incubation in plasma, ANF-(Ser-99-Tyr-126) was partially converted into ANF-(Ser-103-Tyr-126), a less potent peptide. Isolated mesenteric-artery preparation appeared to degrade ANF in a manner very similar to the system in vivo. These results suggest that degradation of ANF may occur either after internalization in the vascular cells or by a membrane-bound enzyme in the vasculature.


Correction of chordee. The Nesbit procedure.
In select patients the Nesbit operation is a useful procedure for the correction of congenital or acquired chordee. This operation results in a deliberate shortening of the convex surface of the corpora cavernosa to counterbalance congenital or acquired shortening on the opposite side. Precise placement of the corrective tuck allows complete correction of the chordee with minimal disturbance of normal tissue. An artificial erection should be created to permit precise identification of the point of maximal curvature on the longer (convex) side of the penis. The transverse elliptical segment to be removed should be placed precisely at this point and should be of sufficient width to result in complete correction of the chordee. Care should be taken to remove only a segment of the tunica albuginea without damaging the underlying erectile tissue. In cases of ventral chordee, the corrective procedure will require elevation of the neurovascular bundle. In cases of dorsal chordee, the procedure will require elevation of the urethra and corpus spongiosum. A repeat artificial erection insures that an adequate correction has been obtained.


Pattern of acute poisonings in childhood in Ankara: what has changed in twenty years?
Poisoning represents one of the most common medical emergencies in childhood, and epidemiological properties differ from country to country. Thus, special epidemiological surveillance for each country is necessary to determine the problem according to which preventive measures can be taken. The purpose of this study was to clarify the characteristics of acute poisoning cases admitted to a pediatric referral hospital. All poisoned patients under 17 years of age, except for cases food poisoning, presenting to the Emergency Department (ED) from January 1995 to December 2000 were determined. The information about each case was recorded on standardized forms and a retrospective chart review survey was done. Complete epidemiological and clinical data were obtained for 489 patients. The mean age of all poisoned patients (mean +/- standard deviation) was 5.96 +/- 4.87 years, and the age range was 0.01 to 17 years. Three hundred and thirty-one children, forming 63.6% of all patients, were under five years of age. Slightly more boys (52.3%) than girls were intoxicated at ages less than 10 years, after which more girls (79%) than boys were involved. The majority of all cases were due to accidental poisoning (78.1% of all poisonings) which occurred mostly in children under five years of age (73.3%). While accidental poisonings (97.1%) were the most common mode of poisoning between 1-5 years, self-poisonings (67.3%) had the highest ratio in cases over 10 years of age. In patients younger than one year of age, 74.2% of all poisonings were due to therapeutical error. Drugs were the most frequent offending agent (57.7%), followed by ingestion of a caustic/corrosive substance (16.8%) and carbon monoxide (CO) intoxication (9.4%). Analgesics were the most common agents, forming 23.7% of all poisonings due to drugs, followed by ingestion of multiple drugs and tricyclic antidepressants at ratios of 21.6% and 9.6%, respectively. The most common route of poisoning was ingestion of the poison (437/489 patients, 89.4%) and most were ingested inside the house (93.3%). About half of all poisoned patients (50.9%) were admitted to the ED within the first two hours of ingestion, and gastric lavage was performed on about half of the poisoned children (48.7%). In most of the cases, hospital treatment was non-specific, including general measures of decontamination and supportive-symptomatic therapy. During the six-year study period, two patients were lost due to acute poisoning, yielding an overall mortality rate of 0.4%. While most of the poisonings were due to accidental ingestions in infancy and primary school ages without sex predilection, the incidence of self-poisonings, especially in girls, was found to be increased. Analgesics, tricyclic antidepressant drugs (which seemed to form a new and dangerous group) and caustic/corrosive substances were the most commonly ingested agents. The early awareness of poisoning and appropriate therapeutic measures taken seemed to be efficacious with a very low mortality rate. The epidemiological and preventive properties of childhood poisonings should be further searched by prospectively designed multicentered studies throughout our country.


Complex I, V, and MDH2 deficient human skin fibroblasts reveal distinct metabolic signatures by 1 H HR-MAS NMR.
In this study, we investigated the metabolic signatures of different mitochondrial defects (two different complex I and complex V, and the one MDH2 defect) in human skin fibroblasts (HSF). We hypothesized that using a selective culture medium would cause defect specific adaptation of the metabolome and further our understanding of the biochemical implications for the studied defects. All cells were cultivated under galactose stress condition and compared to glucose-based cell culture condition. We investigated the bioenergetic profile using Seahorse XFe96 cell analyzer and assessed the extracellular metabolic footprints and the intracellular metabolic fingerprints using NMR. The galactose-based culture condition forced a bioenergetic switch from a glycolytic to an oxidative state in all cell lines which improved overall separation of controls from the different defect groups. The extracellular metabolome was discriminative for separating controls from defects but not the specific defects, whereas the intracellular metabolome suggests CI and CV changes and revealed clear MDH2 defect-specific changes in metabolites associated with the TCA cycle, malate aspartate shuttle, and the choline metabolism, which are pronounced under galactose condition.


Integration of medical imaging into a multi-institutional hospital information system structure.
The Department of Veterans Affairs (VA) is providing integrated text and image data to its clinical users at its Washington and Baltimore medical centers and, soon, at nine other medical centers. The DHCP Imaging System records clinically significant diagnostic images selected by medical specialists in a variety of departments, including cardiology, gastroenterology, pathology, dermatology, surgery, radiology, podiatry, dentistry, and emergency medicine. These images, which include color and gray scale images, and electrocardiogram waveforms, are displayed on workstations located throughout the medical centers. Integration of clinical images with the VA's electronic mail system allows transfer of data from one medical center to another. The ability to incorporate transmitted text and image data into on-line patient records at the collaborating sites is an important aspect of professional consultation. In order to achieve the maximum benefits from an integrated patient record system, a critical mass of information must be available for clinicians. When there is also seamless support for administration, it becomes possible to re-engineer the processes involved in providing medical care.


LETM1 overexpression is correlated with the clinical features and survival outcome of breast cancer.
Leucine zipper/EF hand-containing transmembrane-1 (LETM1) is a mitochondrial inner membrane protein that was first identified in Wolf-Hirschhorn syndrome. However, high-level expression of LETM1 has been correlated with multiple human malignancies, suggesting roles in carcinogenesis and tumor progression. This study is aimed to explore the clinicopathological characteristics and prognostic value of LETM1 overexpression in breast cancer. Immunohistochemical (IHC) staining, and immunofluorescence (IF) were performed to examine LETM1 expression in breast cancer cell line/tissues compared with adjacent normal tissues. Statistical analysis was applied to evaluate the correlation between LETM1 overexpression and the clinicopathological features of breast cancer. Survival rates were calculated using the Kaplan-Meier method, and the relationship between prognostic factors and patient survival was analyzed using the Cox proportional hazard models. LETM1 protein showed cytoplasmic staining pattern in breast cancer. The strongly positive rate of LETM1 protein was 61.6% (98/159) in breast cancer, which was significantly higher than in DCIS (29.7%, 11/37), hyperplasia (16.7%, 3/18) and adjacent normal breast tissues (15.9%, 7/44). High-level expression of LETM1 protein was correlated with lymph node metastasis, poor differentiation, late clinical stage, disease-free survival (DFS) and overall survival (OS) rates in breast cancer. Moreover, multivariate analysis suggested that LETM1 emerged as a significant independent prognostic factor along with clinical stage of patients with breast cancer. LETM1 plays an important role in the progression of breast cancer. High level expression of LETM1 is an independent poor prognostic factor of breast cancer.


Model Electrochemical Biosensor for the Detection of Methanol in Aqueous Solutions with Yeast Cells.
An electrochemical device that serves as a model biosensor and contains yeast Saccharomyces cerevisiae as the active biological element was developed. Different configurations of the electrochemical cells were assembled and tested. Stainless steel was used in the electrochemical cell composition process and the surface of this metal electrode was modified with a thin layer of WO3 if necessary. The yeast Saccharomyces cerevisiae was adhered to the working electrode. The resulting model biosensor was then used to monitor the response to a 10% CH3OH. For detection of biological activity, the electrochemical impedance spectroscopy (EIS) method was applied with a portable potentiostat/galvanostat, where the Bode and the Nyquist plots were interpreted. The stability of the device was beforehand determined by measuring the open circuit potential (OCP). The topography of the electrodes was inspected using the techniques of scanning electron microscopy and optical microscopy. The investigated model biosensor serves as a case study for the development of more complex biosensors that utilize living cells as the active layer.


[Alcoholic liver disease: current state of the problem].
The literature review gives the present views of the diagnosis and treatment of alcoholic liver disease (ALD) and data on the genetic markers associated with the development of ALD and alcohol addiction. It discusses in detail the mechanisms of liver damage within the disease in question and substantiates indications for the use of prednisolone and pentoxifylline in alcoholic hepatitis. Therapeutic approaches to treating different forms of ALD are considered.


[The pathomorphology and pathogenetic problems of liver involvement in icterohemorrhagic leptospirosis].
Morphologic findings on the liver during different time periods of Leptospira infection suggest the leading role in the pathogenesis of the damaged organ early in the course of the disease of a toxic vascular affection of the microcirculatory bed presenting with hyperpermeability of the vascular wall, interstitial edema, dyscomplexation of the liver crosspieces together with inflow of bile to the sinusoidal capillaries. To a lesser extent jaundice is related to the hepatocyte dystrophic and necrotic changes as evidenced by the absence of marked disturbances in the protein-synthetizing function of the liver and low enzymatic activity. At week 2 to 3 of the illness against the background of diminution of the edema and partial or complete restoration of the liver constitution (especially so in less afflicted peripheral portions of the lobes) jaundice with high bilirubinemia is caused by a toxic affliction of hepatocytes and advancing centrolobular cholestasis. High values for bilirubin might be related not only to grave alternative changes in hepatocytes but to microcirculatory abnormalities as well, which fact accounts for the absence in leptospirosis of a direct relationship of the blood level of bilirubin to the degree of affliction of the hepatic cells.


Mental health service utilization among Korean elders in Korean churches: preliminary findings from the Memory and Aging Study of Koreans in Maryland (MASK-MD).
Korean Americans (KA) comprise the fourth largest Asian-American subgroup, with a population estimated at nearly 1.7 million, and the vast majority (up to 85%) of KA elders attends ethnic churches. Despite the rapid increase of the KA elderly population, data on mental health service utilization among KA elders are scarce. Based on a cluster sampling method, the Memory and Aging Study among Koreans in Maryland (MASK-MD) recruited and assessed 630 KA elders (mean age: 70.9 ± 6.1 years; 68.9% female) in KA churches for depression, dementia, and level of mental health service utilization. The Korean versions of the Patient Health Questionnaire (PHQ-9K) and Mini-mental Status Examination (MMSE-KC) were administered by trained community health workers. Of the 630 participants, 23.2% and 7.3% had PHQ-9 scores of 5 ('mild depression') or above and 10 or above ('clinical depression'), respectively. In addition, 7.0% scored below the age- and education-specific cutoff values for probable dementia based on the MMSE-KC. Of the 92 participants with 'clinical depression' or having thoughts of death or self-injury, only 16 (17%) reported utilizing mental health services. Likewise, of 56 participants with probable dementia, only 3 (7.3%) sought treatment from a health care provider. The prevalence of depression and cognitive impairment are high in community-dwelling KA elders attending KA churches, but the rate of mental health service utilization among depressed or cognitively impaired Korean elders is low. Further research is warranted to identify barriers to and strategies for adequate mental health care for Korean immigrant elders.


Individual nutrition therapy and exercise regime: a controlled trial of injured, vulnerable elderly (INTERACTIVE trial).
Proximal femoral fractures are amongst the most devastating consequences of osteoporosis and injurious accidental falls with 25-35% of patients dying in the first year post-fracture. Effective rehabilitation strategies are evolving however, despite established associations between nutrition, mobility, strength and strength-related functional outcomes; there has been only one small study with older adults immediately following fragility fracture where a combination of both exercise and nutrition have been provided. The aim of the INTERACTIVE trial is to establish whether a six month, individualised exercise and nutrition program commencing within fourteen days of surgery for proximal femur fracture, results in clinically and statistically significant improvements in physical function, body composition and quality of life at an acceptable level of cost and resource use and without increasing the burden of caregivers. This randomised controlled trial will be performed across two sites, a 500 bed acute hospital in Adelaide, South Australia and a 250 bed acute hospital in Sydney, New South Wales. Four hundred and sixty community-dwelling older adults aged > 70 will be recruited after suffering a proximal femoral fracture and followed into the community over a 12-month period. Participants allocated to the intervention group will receive a six month individualised care plan combining resistance training and nutrition therapy commencing within 14 days post-surgery. Outcomes will be assessed by an individual masked to treatment allocation at six and 12 months. To determine differences between the groups at the primary end-point (six months), ANCOVA or logistic regression will be used with models adjusted according to potential confounders. The INTERACTIVE trial is among the first to combine nutrition and exercise therapy as an early intervention to address the serious consequence of rapid deconditioning and weight loss and subsequent ability to regain pre-morbid function in older patients post proximal femoral fracture. The results of this trial will guide the development of more effective rehabilitation programs, which may ultimately lead to reduced health care costs, and improvements in mobility, independence and quality of life for proximal femoral fracture sufferers. Australian Clinical Trials Registry: ACTRN12607000017426.


Comparison of kinetic models for analysis of pyruvate-to-lactate exchange by hyperpolarized 13 C NMR.
The activity of specific enzyme-catalyzed reactions may be detected in vivo by (13) C NMR of hyperpolarized (HP) substrates. The signals from HP substrates and products, acquired over time, have been fitted to a number of different mathematical models to determine fluxes, but these models have not been critically compared. In this study, two-pool and three-pool first-order models were constructed to measure flux through lactate dehydrogenase in isolated glioblastoma cells by NMR detection of lactate and pyruvate following the addition of HP [1-(13) C]pyruvate. Mass spectrometry (MS) was used to independently monitor (13) C enrichment in intra- and extracellular lactate. Six models were evaluated using time-dependent pyruvate C2 and lactate C1 HP NMR data acquired by the use of selective excitation pulses, plus (13) C enrichment data from intracellular and extracellular lactate measured by MS. A three-pool bidirectional model provided the most accurate description of pyruvate metabolism in these cells. With computed values for T(1) of pyruvate and lactate, as well as the effect of pulsing, the initial flux through lactate dehydrogenase was well determined by both the two-pool bidirectional and unidirectional models when only HP data were available. The three-pool model was necessary to fit the combined data from both MS and HP, but the simpler two-pool exchange model was sufficient to determine the (13) C lactate concentration when the lactate appearance was measured only by HP.


Mutant p53 can delay growth arrest and loss of CDK2 activity in senescing human fibroblasts without reducing p21(WAF1) expression.
Functional wild-type p53 is required for human diploid fibroblasts (HDF) to enter an irreversible growth arrest known as replicative senescence. Experimentally, abrogation of p53 function by expression of human papillomavirus type 16 E6 or disruption of a key downstream effector p21 by homologous recombination both extended HDF life span. However, although sufficient to extend life span, p21 down-regulation is not necessary, because expression of a dominant-negative mutant p53 (143(ala)) extends life span without apparently decreasing p21 expression. Given the importance of p53 in cellular senescence and the general assumption that p21 may be the sole mediator of its action in this process, we have investigated how abrogation of p53 function can overcome senescence without lowering expression of p21. We have found up-regulated levels of the cyclin-dependent kinase 2 (cdk2) protein in HDF expressing 143(ala) mutant p53 as compared to senescent controls, together with an increase in p21-free cdk2 which, in conjunction with cyclin E, is able to form an active kinase which can phosphorylate the retinoblastoma protein. However, forced overexpression of cdk2 in near-senescent HDF failed to restore cdk2-associated kinase activity. Our data suggest that p53-mediated senescence depends on factor(s) other than p21 which modulate formation of cyclin E-cdk2 complexes.


Pitfalls of predicting complex traits from SNPs.
The success of genome-wide association studies (GWASs) has led to increasing interest in making predictions of complex trait phenotypes, including disease, from genotype data. Rigorous assessment of the value of predictors is crucial before implementation. Here we discuss some of the limitations and pitfalls of prediction analysis and show how naive implementations can lead to severe bias and misinterpretation of results.


[Transcranial Doppler ultrasound--robotics-controlled transducers for quantifying the effect of acupuncture].
Transcranial Doppler Sonography (TCD) monitoring was carried out in 12 healthy subjects (mean age: 35.2 +/- 4.4 years; range: 26-41 years; 4 women, 8 men), prior to, during and following acupuncture. In all subjects an identical acupuncture scheme with a "general increase in Qi energy" setting was employed. Measurements were obtained with new robotic TCD probes enabling three-dimensional imaging of the middle cerebral artery by the use of the multi-scan technique. The results showed a significant (p < 0.01; ANOVA; Tukey test) increase in mean bloodflow velocity measured simultaneously at different depths within the right middle cerebral artery. Quantifiable changes in the physiological effects of acupuncture may contribute to improved acceptance of this unconventional complementary medical procedure.


Radiomics model based on multi-sequence MRI for preoperative prediction of ki-67 expression levels in early endometrial cancer.
To validate a radiomics model based on multi-sequence magnetic resonance imaging (MRI) in predicting the ki-67 expression levels in early-stage endometrial cancer, 131 patients with early endometrial cancer who had undergone pathological examination and preoperative MRI scan were retrospectively enrolled and divided into two groups based on the ki-67 expression levels. The radiomics features were extracted from the T2 weighted imaging (T2WI), dynamic contrast enhanced T1 weighted imaging (DCE-T1WI), and apparent diffusion coefficient (ADC) map and screened using the Pearson correlation coefficients (PCC). A multi-layer perceptual machine and fivefold cross-validation were used to construct the radiomics model. The receiver operating characteristic (ROC) curves analysis, calibration curves, and decision curve analysis (DCA) were used to assess the models. The combined multi-sequence radiomics model of T2WI, DCE-T1WI, and ADC map showed better discriminatory powers than those using only one sequence. The combined radiomics models with multi-sequence fusions achieved the highest area under the ROC curve (AUC). The AUC value of the validation set was 0.852, with an accuracy of 0.827, sensitivity of 0.844, specificity of 0.773, and precision of 0.799. In conclusion, the combined multi-sequence MRI based radiomics model enables preoperative noninvasive prediction of the ki-67 expression levels in early endometrial cancer. This provides an objective imaging basis for clinical diagnosis and treatment.


Double Reading of Outsourced CT/MR Radiology Reports: Retrospective Analysis.
Our objective was to determine disagreement rates in radiological reports provided by using a double-reading protocol in a national teleradiology company. From January 2015 to July 2016, 134169 radiological exams from 36 French centers, benefited outsourced interpretations by certified radiologists, in both regular and after-hours activities. Of these, 2040 CT and MR-scans (1.5%) were subjected to a second opinion by other radiologists in the field of their anatomical specialty (cerebral, thoracic, abdominal-pelvic, and osteoarticular). A five-point agreement scale graded from 0 to 4 was assigned for each exam. Disagreements were considered as minor if no clinical consequence for patient (scores 1 and 2) and major if potential clinical consequence (score 3 and 4). Independent radiologists performed a retrospective analysis and a stratified statistical analysis. Double reading was performed on CT-scans (n = 934/2040, 45.8%) and MR-scans (n = 1106/2040, 54.2%) performed in regular (80.1%) and after-hours activities (19.9%). Disagreement scores occurred in 437 exams (21.4%), including major disagreements in 59 (2.9%). Among these, 48/754 were assigned by the thoracic second reader (6.4%), 6/70 by the abdominal-pelvic second reader (8.6%), 3/901 by the osteoarticular second reader (0.3%), and 2/315 by the cerebral second reader (0.6%), with statistical significant difference. No additional disagreement rate was observed in regular and after-hours activities (P = 0.63). Double-reading of outsourced CT and MRI interpretations yielded 21.4% disagreement rate, with potential clinical consequence for patient in 2,9% of the cases. These results are in accordance with those previously reported and suggests that quality assurance of outsourced interpretations is needed.


A clinician's guide to X-linked hypophosphatemia.
X-linked hypophosphatemia (XLH) is the prototypic disorder of renal phosphate wasting, and the most common form of heritable rickets. Physicians, patients, and support groups have all expressed concerns about the dearth of information about this disease and the lack of treatment guidelines, which frequently lead to missed diagnoses or mismanagement. This perspective addresses the recommendation by conferees for the dissemination of concise and accessible treatment guidelines for clinicians arising from the Advances in Rare Bone Diseases Scientific Conference held at the NIH in October 2008. We briefly review the clinical and pathophysiologic features of the disorder and offer this guide in response to the conference recommendation, based on our collective accumulated experience in the management of this complex disorder.


The ontogeny of brain receptors for corticotropin-releasing factor and the development of their functional association with adenylate cyclase.
This study reports the ontogeny of corticotropin-releasing factor (CRF) receptor binding sites in rat brain, using both membrane binding assays and in vitro receptor autoradiography. CRF binding sites are evident by prenatal day 17, increase to 312% of their adult density by postnatal day 8, then decrease to reach adult values by day 21. Not only the density, but the distribution of CRF binding undergoes major modifications in development. CRF binding sites are most numerous in striatum prenatally, but postnatally, binding is more dense in the cortex, reaching the adult laminar distribution by postnatal day 14. Brain CRF receptors are linked to adenylate cyclase early in postnatal life. This contrasts with the later appearance of most of the guanine nucleotide stimulatory protein and catalytic subunit sites in the rat brain and suggests that CRF receptors may become functional earlier than several other brain receptors that are linked to adenylate cyclase.


Acquired immunodeficiency syndrome (AIDS) in homosexual men--a new public health concern.
The acquired immunodeficiency syndrome (AIDS) is a new, serious condition of major public health concern. It is most prevalent among young homosexuals and may be present with Kaposi's sarcoma and serious opportunistic infections. Physicians dealing with male homosexuals should be aware of this diagnostic possibility in any patient with unusual skin lesions or unusual clinical findings indicating infection. AIDS may be preceded by persistent generalized lymphadenopathy which may be accompanied by fatigue, low grade fever, night sweats and weight loss. The prognosis is poor with high mortality rates and no indication of spontaneous recovery of cellular immunocompetence.