A radical-chemical route to acetyl-CoA: the anaerobically induced pyruvate formate-lyase system of Escherichia coli.
Anaerobically growing Escherichia coli cells contain the enzyme pyruvate formate-lyase which catalyses the non-oxidative cleavage of pyruvate to acetyl-CoA and formate. The enzyme is subject to interconversion between inactive and active forms. The active form contains an oxygen-sensitive organic free radical located on the polypeptide chain which is essential for catalysis. It affords a novel homolytic C-C bond cleavage of the pyruvate substrate. The radical is generated by an iron-dependent converter enzyme which requires reduced flavodoxin and adenosyl methionine as co-substrates and pyruvate as a positive allosteric effector. A second converter enzyme, also iron-dependent, accomplishes the removal of the radical. This post-translational interconversion cycle controls the activity state of pyruvate formate-lyase in the anaerobic cell. Anaerobic conditions also regulate pyruvate formate-lyase at the level of gene expression. Multiple promoters are responsible for effecting a twelve to fifteen fold induction and they are coordinately controlled in response to the oxygen and metabolic status of the cell by sequences which are located far upstream of the pfl coding region. The transcription factor Fnr has been identified as being responsible for part of the anaerobic control of pfl expression, probably through direct interaction with the upstream sequences. In contrast, the expression of the gene encoding the first iron-dependent converter enzyme is unaffected by anaerobiosis and is independent of the Fnr protein.


[Subjective body experience and therapy of somatoform disorders].
The article addresses the problem of subjective body experience in patients with somatoform mental disorders. Perspectives of multi-axial model of subjective body experience and methodical approaches to its descriptions are discussed. Different variants of body experience organization are described. Clinical peculiarities of somatoform mental disorders were closely connected with the complicated structure of body experience. Main processes involving body experience are those of self-identification structuring: determination of the borders of subjectivity, relations with objects, representation of self-ego. An analysis of structure of subjective body experience enabled to work out differentiated psychotherapeutic approaches to complex rehabilitation of patients with somatoform mental disorders.


Malnutrition as a predictor of poor postoperative outcomes in gynecologic cancer patients.
Poor nutritional status has been associated with increased postoperative morbidity and mortality in surgical patients. The purpose of this study is to evaluate if decreased nutritional parameters correlate with increased postoperative complications regardless of other risk factors in the gynecologic cancer patient. A retrospective chart review was performed among women who underwent surgical management for gynecologic malignancies from October 2006 to June 2008. Variables included age, race, medical comorbidities, cancer type/stage, preoperative albumin, absolute lymphocyte count (ALC), and body mass index (BMI), estimated blood loss (EBL), intraoperative blood transfusion (BT), intraoperative or postoperative complications, intensive care unit (ICU) admissions, hospital readmissions, reoperations, and cancer recurrence. Three hundred gynecologic oncology patients with preoperative nutritional parameters were included in the study. Decreased albumin was significantly associated with more postoperative complications (p < 0.001), hospital readmissions (p = 0.01), reoperations (p = 0.03), ICU admissions (p < 0.001), and cancer recurrence (p < 0.001). Decreased ALC and BMI preoperatively was also significantly associated with higher incidence of cancer recurrence (p = 0.01, p = 0.01). Surgical cases involving increased EBL (p = 0.01, p < 0.001) and more BT (p < 0.001, p < 0.001) had significantly more postoperative complications and more ICU admissions. Multivariable logistic regression found preoperative albumin to be an independent predictor of increased postoperative complications. Decreased albumin is significantly associated with more postoperative complications, hospital readmissions, reoperations, ICU admissions, and cancer recurrence. This nutritional parameter is an important predictor of postoperative morbidity and mortality. Thus, it is important to assess nutritional status preoperatively and offer nutritional support or alternate treatment options if necessary.


Expression of the ET(A) and ET(B) endothelin receptor subtype mRNA in human detrusor cultured smooth muscle cells.
The aim of this study was to determine the expression of the endothelin receptor subtype mRNAs in human detrusor cultured smooth muscle cells using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). First strand cDNA was made from human detrusor cultured smooth muscle cells total RNA and used for PCR with primers designed to amplify fragments of the ET(A) and ET(B) endothelin receptor subtype cDNA sequences. Subcloned fragments of the ET(A) and ET(B) endothelin receptor cDNAs were used to synthesize digoxigenin-labeled cRNA probes by in vitro transcription. COS-7 cells transfected with the ET(A) and ET(B) receptor cDNAs were used as positive control and to confirm the absence of cross-hybridization due to sequence homology. Both ET(A) and ET(B) receptor mRNAs were detected by RT-PCR analysis. By ISH, both ET(A) and ET(B) receptor subtype mRNAs were detected. However, ET(A) signal was much more intense than ET(B) signal. These results indicate that mRNAs for both ET(A) and ET(B) receptors are expressed in detrusor smooth muscle cells of human urinary bladder. The ET(A) receptor is the predominant detrusor ET receptor.


Radiographic evaluation of hip resurfacing: the role of x-rays in the diagnosis of a problematic resurfaced hip.
Hip resurfacing (HRA) requires a new standardised radiographic evaluation protocol. Evaluation of acetabular components is similar to total hip arthroplasty (THA) but femoral components require different criteria. This study evaluates the efficacy of a new femoral zonal system for radiographic HRA assessment in identifying clinical problems. A new proximal femoral zonal system divides implant-cement-bone interfaces at head and neck into 7 DeSmet zones. 611 in situ and 100 revised HRA were assessed for component positioning, borderline findings (reactive lines, cortical thickening, cancellous condensation) or sinister findings (lucent lines, bone resorption, osteolysis). Findings were correlated with gender, size, implant survival, clinical scores, metal ions, and adverse soft tissue reactions (ALTR). Radiological changes were found in 265 hips (37.3%), 154 sinister (21.7%) and 111 borderline (15.6%). 84% of revisions had sinister findings versus 11.4% of in situ HRA (p < 0.001). The mean number of abnormal acetabular and femoral zones was 3.3 in revised compared to 0.6 in situ HRA (p < 0.001). Sinister findings were associated with female gender, lower clinical scores, smaller sizes or ASR design (all: p < 0.001). Metal ions were higher with sinister (p < 0.001) and borderline (p < 0.01) findings compared to normal radiographs. Pathological findings in ⩾4 zones had a sensitivity of 50% and specificity of 94.2% (AUC = 0.898) in detecting hips with ALTR (odds ratio = 49). In order to accurately evaluate progressive radiographic changes in HRA, specific zones have been established around the femoral component. There was a high correlation between radiographic findings and outcome measures. 84% of problematic hips and 90.4% of proven ALTR had sinister changes.


Aspiration of thoracic masses and fluid collections under guidance of ultrasonography.
Successful ultrasonic detection and fine needle aspiration of pleural effusions, loculated fluid, and peripheral lung masses were done in 115 patients. No complications, eg, pneumothorax or hematoma, were caused by the fine needle aspirations. There was no false-negative or false-positive cytology because of excellent localization by ultrasound and adequate tissue samples. This series shows ultrasonography to be a valid method of evaluating chest lesions seen to be peripheral on chest roentgenograms. The cystic or solid nature of these lesions was determined; however, we experienced difficulty in differentiating small amounts of loculated fluid from pleural thickening in a few cases. Difficulties were also noted with a few small peripheral tumors that appeared to be cystic on ultrasonography but were proven to be solid by fine needle aspiration. These problems can be resolved by an experienced sonographer using equipment with higher resolution, but we did not hesitate to do fine needle aspiration in such circumstances.


[Acute hyperparathyroidism caused by hemorrhage into a parathyroid adenoma].
Acute hyperparathyroidism is a rare and dangerous condition. It is supposed to result from transformation of a mild chronic form into an acute and stormy illness. The causes and the mechanism of such a transformation are usually unknown. A case of acute hyperparathyroidism is presented in which hemorrhage into a parathyroid adenoma was recognized and diagnosed as the immediate cause of the transformation from the mild long-standing form of the disease into the severe acute form.


Levels and fate of perfluoroalkyl substances in beached plastic pellets and sediments collected from Greece.
Plastic debris damages marine wildlife and ecosystems becoming an important source of marine pollution. In addition, they can sorb, concentrate and stabilise contaminants acting as toxic carriers to the marine food web. In this context, the presence of 18 perfluoroalkyl substances (PFASs) in plastic pellets (n=5) and beach sediment (n=9) samples widely distributed around Greek coastal areas was assessed. The results, mainly, showed the sorption of PFASs onto pellet surface from surrounding water with concentrations from method limit of quantification to 115 ng/kg for C5, C7, C8 and C10 carboxylic acids and C8 sulfonate acid. A similar pattern was found by comparing plastic pellets and sediment for the same sampling locations that could indicate a common origin of contamination in both types of samples. However, since the number of analysed samples is limited, a more comprehensive study with a higher number of samples should be performed in future research.


Protein substitute for children and adults with phenylketonuria.
Phenylketonuria is an inherited metabolic disorder characterised by an absence or deficiency of the enzyme phenylalanine hydroxylase. The aim of treatment is to lower blood phenylalanine concentrations to the recommended therapeutic range to prevent developmental delay and support normal growth. Current treatment consists of a low-phenylalanine diet in combination with a protein substitute which is free from or low in phenylalanine. Guidance regarding the use, dosage, and distribution of dosage of the protein substitute over a 24-hour period is unclear, and there is variation in recommendations among treatment centres. This is an update of a Cochrane review first published in 2005, and previously updated in 2008. To assess the benefits and adverse effects of protein substitute, its dosage, and distribution of dose in children and adults with phenylketonuria who are adhering to a low-phenylalanine diet. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register which consists of references identified from comprehensive electronic database searches and hand searches of relevant journals and abstract books of conference proceedings. We also contacted manufacturers of the phenylalanine-free and low-phenylalanine protein substitutes for any data from published and unpublished randomised controlled trials.Date of the most recent search of the Group's Inborn Errors of Metabolism Trials Register: 03 April 2014. All randomised or quasi-randomised controlled trials comparing: any dose of protein substitute with no protein substitute; an alternative dosage; or the same dose, but given as frequent small doses throughout the day compared with the same total daily dose given as larger boluses less frequently. Both authors independently extracted data and assessed trial quality. Three trials (69 participants) are included in this review. One trial investigated the use of protein substitute in 16 participants, while a further two trials investigated the dosage of protein substitute in a total of 53 participants. Due to issues with data presentation in each trial, described in full in the review, formal statistical analyses of the data were impossible. Investigators will be contacted for further information. No conclusions could be drawn about the short- or long-term use of protein substitute in phenylketonuria due to the lack of adequate or analysable trial data. Additional data and randomised controlled trials are needed to investigate the use of protein substitute in phenylketonuria. Until further evidence is available, current practice in the use of protein substitute should continue to be monitored with care.


Image Fusion Involving Real-Time Transabdominal or Endoscopic Ultrasound for Gastrointestinal Malignancies: Review of Current and Future Applications.
Image fusion of CT, MRI, and PET with endoscopic ultrasound and transabdominal ultrasound can be promising for GI malignancies as it has the potential to allow for a more precise lesion characterization with higher accuracy in tumor detection, staging, and interventional/image guidance. We conducted a literature review to identify the current possibilities of real-time image fusion involving US with a focus on clinical applications in the management of GI malignancies. Liver applications have been the most extensively investigated, either in experimental or commercially available systems. Real-time US fusion imaging of the liver is gaining more acceptance as it enables further diagnosis and interventional therapy of focal liver lesions that are difficult to visualize using conventional B-mode ultrasound. Clinical studies on EUS guided image fusion, to date, are limited. EUS-CT image fusion allowed for easier navigation and profiling of the target tumor and/or surrounding anatomical structure. Image fusion techniques encompassing multiple imaging modalities appear to be feasible and have been observed to increase visualization accuracy during interventional and diagnostic applications.


Calcium ion mechanisms in airway smooth muscle: potential targets for novel symptomatic drugs for asthma.
Calcium ions (Ca2+) are fundamental to the processes responsible for the initiation and maintenance of contraction of airway smooth muscle cells. Recent developments in our understanding of signal transduction mechanisms relating to intracellular Ca2+ release have extended our knowledge of excitation-contraction coupling mechanisms in airway smooth muscle. Furthermore, these developments open up potential targets for the development of new drugs, with novel mechanisms of action, for the symptomatic treatment of asthma. This article reviews these recent advances and focusses upon those calcium ion mechanisms that are possible targets for drug action.


A lipidomic platform establishment for structural identification of skin ceramides with non-hydroxyacyl chains.
The stratum corneum (SC) is the outermost layer of skin that functions as a barrier and protects against environmental influences and transepidermal water loss. Its unique morphology consists of keratin-enriched corneocytes embedded in a distinctive mixture of lipids containing mainly ceramides, free fatty acids, and cholesterol. Ceramides are sphingolipids consisting of sphingoid bases, which are linked to fatty acids by an amide bond. Typical sphingoid bases in the skin are composed of dihydrosphingosine (dS), sphingosine (S), phytosphingosine (P), and 6-hydroxysphingosine (H), and the fatty acid acyl chains are composed of non-hydroxy fatty acid (N), α-hydroxy fatty acid (A), ω-hydroxy fatty acid (O), and esterified ω-hydroxy fatty acid (E). The 16 ceramide classes include several combinations of sphingoid bases and fatty acid acyl chains. Among them, N-type ceramides are the most abundant in the SC. Mass spectrometry (MS)/MS analysis of N-type ceramides using chip-based direct infusion nanoelectrospray-ion trap mass spectrometry generated the characteristic fragmentation pattern of both acyl and sphingoid units, which could be applied to structural identification of ceramides. Based on the MS/MS fragmentation patterns of N-type ceramides, comprehensive fragmentation schemes were proposed. In addition, mass fragmentation patterns, which are specific to the sphingoid backbone of N-type ceramides, were found in higher m/z regions of tandem mass spectra. These characteristic and general fragmentation patterns were used to identify N-type ceramides in human SC. Based on established MS/MS fragmentation patterns of N-type ceramides, 52 ceramides (including different classes of NS, NdS, NP, and NH) were identified in human SC. The MS/MS fragmentation patterns of N-type ceramides were characterized by interpreting their product ion scan mass spectra. This information may be used to identify N-type ceramides in the SC of human, rat, and mouse skin.


The necessary and the desirable production of radicals in biology.
The production of radicals within biological systems can be necessary, as in the coupling of light capture or oxygen-hydride reactions to the generation of proton gradients in thylakoid and mitochondrial membranes; or it can be extremely desirable, to produce an effective catalyst centre in, for example, ribonucleotide reductases. To fit their function the radicals are usually of controlled reactivity, either through their redox properties or through confinement to a limited volume. The more indiscriminate generation of radicals can be of great use too in protective devices, for example, in the production of cross-linked polymers, or in the generation of oxygen radicals. The different distribution and release of initiating enzymes for all these radical processes ensures the spatial and temporal control of their reactions.


Postincident review of aggression and violence in mental health settings.
Management of violence and aggression remains a challenge to mental health care practitioners. It has been acknowledged that for a small number of incidents involving aggression, use of restraint will continue to be a method of containing potentially dangerous situations. The impact of being involved in these procedures remains under researched, but there is growing acknowledgment that some form of post incident review should take place after restraint use. As part of a larger study, a survey design was used to evaluate whether staff (n = 30) and inpatients (n = 30) had found post incident review helpful after incidents involving restraint. Ninety-seven percent of staff, and 94% of patient participants agreed this approach was useful. This article presents the findings of this survey and discusses the complex factors that should be considered when reviewing the aftermath of restraint for staff and inpatients in acute mental health settings.


Validation of an HPLC method for the determination of urinary and plasma levels of N1-methylnicotinamide, an endogenous marker of renal cationic transport and plasma flow.
N1-Methylnicotinamide (NMN) is an endogenous cationic metabolite of nicotinamide (niacine, vitamine PP) whose renal clearance reflects both the capacity of the renal tubular transport system to secrete organic cations and renal plasma flow. NMN is present in human plasma and urine at the 1-117-ng ml(-1) and 0.5-25-microg ml(-1) concentration range, respectively, and its level depends notably on pathophysiological (age, renal or hepatic diseases) conditions. We report the optimization and validation of an HPLC method for the measurement of endogenous NMN in biological fluids after derivatization into a fluorescent compound. Plasma is first deproteinized with TCA 20% and the urine diluted 1:10 with HCI 10(-4) M prior to the derivatization procedure, which includes a condensation reaction of NMN with acetophenone in NaOH at 0 degrees C, followed by dehydration in formic acid and subsequent formation of the fluorescent 1,6-naphthyridine derivatives after heating samples in a boiling water bath. The synthetic homologous derivative N1-ethylnicotinamide (NEN) reacts similarly and is added as internal standard into the biological fluid. The reaction mixture is subjected to reverse phase high performance liquid chromatography on a Nucleosil 100-C18 column using a mobile phase (acetonitrile 22%, triethylamine 0.5%, 0.01 M sodium heptanesulfonate adjusted to pH 3.2), delivered isocratically at a flow rate of 1 ml min(-1), NMN and NEN are detected at 7.8 and 10 min by spectrofluorimetry with excitation and emission wavelengths set at 366 and 418 nm, respectively. The addition-calibration method is used with plasma and urine pools. Calibration curves (using the internal standard method) are linear (r2 > 0.997) at concentrations up to 109 ng ml(-1) and 15.7 microg ml(-1) in plasma and urine, respectively. Both intra- and inter-assay precision of plasma control samples at 10, 50 and 90 ng ml(-1) were lower than 3.3% and concentrations not deviating more than 2.7% from their nominal values. In urine intra- and inter-assay CVs of control samples at 1, 5 and 9 microg ml(-1) are lower than 8.3%, with concentrations not deviating more than -9.0 to +11.8% from their nominal values. This analytical method has therefore the required sensitivity and selectivity to measure NMN in plasma and urine, enabling the non-invasive determination of the tubular secretory capacity of the kidney and the renal plasma flow.


A short screening instrument for poststroke dementia : the R-CAMCOG.
The CAMCOG is a feasible cognitive screening instrument for dementia in patients with a recent stroke. A major disadvantage of the CAMCOG, however, is its lengthy and relatively complex administration for screening purposes. We therefore developed the Rotterdam CAMCOG (R-CAMCOG), based on the original version. Our aim was to reduce the estimated administration time to 15 minutes or less and to retain or perhaps even improve its diagnostic accuracy. We analyzed the item scores on the CAMCOG of 300 consecutive stroke patients, after exclusion of patients with a severe aphasia or lowered consciousness level, who were entered in the Rotterdam Stroke Databank. The diagnosis of dementia was made independent of the R-CAMCOG score, on the basis of clinical examination and neuropsychological test results. The R-CAMCOG was constructed in 3 steps. First, items with floor and ceiling effects were removed. Next, subscales with no additional diagnostic value were excluded. Finally, we removed items that did not contribute to the homogeneity of the subscales. The diagnostic accuracy of the R-CAMCOG and the original CAMCOG was determined by means of the area under the receiver operating characteristic (ROC) curve. In the 3 steps, the number of items was reduced from 59 to 25, divided over the subscales orientation, memory (recent, remote, and learning), perception, and abstraction. The subscale orientation did not reach significance in a logistic regression model but was included in the R-CAMCOG because of its high face validity in dementia screening. Internal validation with ROC analysis suggests that the R-CAMCOG and the CAMCOG are equally accurate in screening for poststroke dementia (area under the curve was 0.95 for both tests). The R-CAMCOG has overcome the disadvantages of the original CAMCOG. It is a promising, short, and easy-to-administer screening instrument for poststroke dementia. It seems to be sufficiently accurate for this purpose, but the test has yet to be validated in a separate, independent study.


White adipose remodeling during browning in mice involves YBX1 to drive thermogenic commitment.
Increasing adaptive thermogenesis by stimulating browning in white adipose tissue is a promising method of improving metabolic health. However, the molecular mechanisms underlying this transition remain elusive. Our study examined the molecular determinants driving the differentiation of precursor cells into thermogenic adipocytes. In this study, we conducted temporal high-resolution proteomic analysis of subcutaneous white adipose tissue (scWAT) after cold exposure in mice. This was followed by loss- and gain-of-function experiments using siRNA-mediated knockdown and CRISPRa-mediated induction of gene expression, respectively, to evaluate the function of the transcriptional regulator Y box-binding protein 1 (YBX1) during adipogenesis of brown pre-adipocytes and mesenchymal stem cells. Transcriptomic analysis of mesenchymal stem cells following induction of endogenous Ybx1 expression was conducted to elucidate transcriptomic events controlled by YBX1 during adipogenesis. Our proteomics analysis uncovered 509 proteins differentially regulated by cold in a time-dependent manner. Overall, 44 transcriptional regulators were acutely upregulated following cold exposure, among which included the cold-shock domain containing protein YBX1, peaking after 24 h. Cold-induced upregulation of YBX1 also occurred in brown adipose tissue, but not in visceral white adipose tissue, suggesting a role of YBX1 in thermogenesis. This role was confirmed by Ybx1 knockdown in brown and brite preadipocytes, which significantly impaired their thermogenic potential. Conversely, inducing Ybx1 expression in mesenchymal stem cells during adipogenesis promoted browning concurrent with an increased expression of thermogenic markers and enhanced mitochondrial respiration. At a molecular level, our transcriptomic analysis showed that YBX1 regulates a subset of genes, including the histone H3K9 demethylase Jmjd1c, to promote thermogenic adipocyte differentiation. Our study mapped the dynamic proteomic changes of murine scWAT during browning and identified YBX1 as a novel factor coordinating the genomic mechanisms by which preadipocytes commit to brite/beige lineage.


Anterior versus posterior approach in reconstruction of infected nonunion of the tibia using the vascularized fibular graft: potentialities and limitations.
The potentialities, limitations, and technical pitfalls of the vascularized fibular grafting in infected nonunions of the tibia are outlined on the basis of 14 patients approached anteriorly or posteriorly. An infected nonunion of the tibia together with a large exposed area over the shin of the tibia is better approached anteriorly. The anastomosis is placed in an end-to-end or end-to-side fashion onto the anterior tibial vessels. To locate the site of the nonunion, the tibialis anterior muscle should be retracted laterally and the proximal and distal ends of the site of the nonunion debrided up to healthy bleeding bone. All the scarred skin over the anterior tibia should be excised, because it becomes devitalized as a result of the exposure. To cover the exposed area, the fibula has to be harvested with a large skin paddle, incorporating the first septocutaneous branch originating from the peroneal vessels before they gain the upper end of the flexor hallucis longus muscle. A disadvantage of harvesting the free fibula together with a skin paddle is that its pedicle is short. The skin paddle lies at the antimesenteric border of the graft, the site of incising and stripping the periosteum. In addition, it has to be sutured to the skin at the recipient site, so the soft tissues (together with the peroneal vessels), cannot be stripped off the graft to prolong its pedicle. Vein grafts should be resorted to, if the pedicle does not reach a healthy segment of the anterior tibial vessels. Defects with limited exposed areas of skin, especially in questionable patency of the vessels of the leg, require primarily a fibula with a long pedicle that could easily reach the popliteal vessels and are thus better approached posteriorly. In this approach, the site of the nonunion is exposed medial to the flexor digitorum muscle and the proximal and distal ends of the site of the nonunion debrided up to healthy bleeding bone. No attempt should be made to strip the scarred skin off the anterior aspect of the bone lest it should become devitalized. Any exposed bone on the anterior aspect should be left to granulate alone. This occurs readily when stability has been regained at the fracture site after transfer of the free fibula. The popliteal and posterior tibial vessels are exposed, and the microvascular anastomosis placed in an end-to-side fashion onto either of them, depending on the length of the pedicle and the condition of the vessels themselves. To obtain the maximal length of the pedicle of the graft, the proximal osteotomy is placed at the neck of the fibula after decompressing the peroneal nerve. The distal osteotomy is placed as distally as possible. After detaching the fibula from the donor site, the proximal part of the graft is stripped subperiosteally, osteotomized, and discarded. Thus, a relatively long pedicle could be obtained. To facilitate subperiosteal stripping, the free fibula is harvested without a skin paddle. In this way, the use of a vein graft could be avoided. Patients presenting with infected nonunions of the tibia with extensive scarring of the lower extremity, excessively large areas of skin loss, and with questionable patency of the anterior and posterior tibial vessels are not suitable candidates for the free vascularized fibular graft. Although a vein graft could be used between the recipient popliteal and the donor peroneal vessels, its use decreases flow to the graft considerably. These patients are better candidates for the Ilizarov bone transport method with or without free latissimus dorsi transfer.


The Management of Opioid-Induced Constipation in Cancer and Advanced Illness: A Meta-Analysis.
Constipation is a common problem among patients with cancer. By some accounts, about 60% of cancer patients experience constipation. There is limited empirical evidence of the clinical effectiveness of pharmacologic agents in opioid-induced constipation in advanced diseases. We sought to quantitatively summarize the therapeutic effectiveness of the pharmacologic means of managing opioid-induced constipation. Randomized control trials (RCTs) identified from medical literature databases that reported quantitative measures of the effect of pharmacotherapeutic agents to treat opioid induced constipation in patients with cancers and other advanced illnesses were included in this study. A conventional random effects meta-analysis was conducted including >3 trials with the same exposure and outcome assessed, and a network-meta-analysis was conducted for all placebo-controlled trials. Eighteen studies that examined the effect of various pharmacotherapeutic agents were included. The medications were Methylnatrexone (N=5), Naldemedine (N=5), other conventional agents (N=4) and herbal medicines (N=4). In conventional meta-analysis, methylnaltrexone increased the proportion achieving rescue-free laxation by 2.68 fold (95% CI: 1.34, 5.37; p=0.0054) within four hours of the administration compared to placebo. In network meta-analysis, the pooled RR of the pharmacotherapeutic agents on rescue-free bowel movements as 2.26 (95% CI: 1.52, 3.36) for methylnaltrexone, 1.58 (95% CI: 0.94, 2.66) for naldemedine, and 0.74 (95% CI: 0.45, 1.23) for polyethylene glycol, compared to placebo. Methylnatrexone and Naldemedine have currently shown promise in randomized trials concerning Opioid-Induced Constipation in Cancer and Advanced Illness. It is imperative that future research ascertain not just the relative therapeutic efficacy but also the cost-benefit analyses of these newer regimens with more commonly used and accessible laxatives.


Recent advances in the histo-molecular pathology of human prion disease.
Prion diseases are progressive neurodegenerative disorders affecting humans and other mammalian species. The term prion, originally put forward to propose the concept that a protein could be infectious, refers to PrPSc , a misfolded isoform of the cellular prion protein (PrPC ) that represents the pathogenetic hallmark of these disorders. The discovery that other proteins characterized by misfolding and seeded aggregation can spread from cell to cell, similarly to PrPSc , has increased interest in prion diseases. Among neurodegenerative disorders, however, prion diseases distinguish themselves for the broader phenotypic spectrum, the fastest disease progression and the existence of infectious forms that can be transmitted through the exposure to diseased tissues via ingestion, injection or transplantation. The main clinicopathological phenotypes of human prion disease include Creutzfeldt-Jakob disease, by far the most common, fatal insomnia, variably protease-sensitive prionopathy, and Gerstmann-Sträussler-Scheinker disease. However, clinicopathological manifestations extend even beyond those predicted by this classification. Because of their transmissibility, the phenotypic diversity of prion diseases can also be propagated into syngenic hosts as prion strains with distinct characteristics, such as incubation period, pattern of PrPSc distribution and regional severity of histopathological changes in the brain. Increasing evidence indicates that different PrPSc conformers, forming distinct ordered aggregates, encipher the phenotypic variants related to prion strains. In this review, we summarize the most recent advances concerning the histo-molecular pathology of human prion disease focusing on the phenotypic spectrum of the disease including co-pathologies, the characterization of prion strains by experimental transmission and their correlation with the physicochemical properties of PrPSc aggregates.


Interaction between effects of genes coding for dopamine and glutamate transmission on striatal and parahippocampal function.
The genes for the dopamine transporter (DAT) and the D-Amino acid oxidase activator (DAOA or G72) have been independently implicated in the risk for schizophrenia and in bipolar disorder and/or their related intermediate phenotypes. DAT and G72 respectively modulate central dopamine and glutamate transmission, the two systems most robustly implicated in these disorders. Contemporary studies have demonstrated that elevated dopamine function is associated with glutamatergic dysfunction in psychotic disorders. Using functional magnetic resonance imaging we examined whether there was an interaction between the effects of genes that influence dopamine and glutamate transmission (DAT and G72) on regional brain activation during verbal fluency, which is known to be abnormal in psychosis, in 80 healthy volunteers. Significant interactions between the effects of G72 and DAT polymorphisms on activation were evident in the striatum, parahippocampal gyrus, and supramarginal/angular gyri bilaterally, the right insula, in the right pre-/postcentral and the left posterior cingulate/retrosplenial gyri (P < 0.05, FDR-corrected across the whole brain). This provides evidence that interactions between the dopamine and the glutamate system, thought to be altered in psychosis, have an impact in executive processing which can be modulated by common genetic variation.


Language and communication development in Down syndrome.
Although there is considerable variability, most individuals with Down syndrome have mental retardation and speech and language deficits, particularly in language production and syntax and poor speech intelligibility. This article describes research findings in the language and communication development of individuals with Down syndrome, first briefly describing the physical and cognitive phenotype of Down syndrome, and two communication related domains-hearing and oral motor skills. Next, we describe language development in Down syndrome, focusing on communication behaviors in the prelinguistic period, then the development of language in children and adolescents, and finally language development in adults and the aging period. We describe language development in individuals with Down syndrome across four domains: phonology, semantics, syntax, and pragmatics. Wethen suggest strategies for intervention and directions for research relating to individuals with Down syndrome.


Plasma proteome analysis reveals the geographical origin and liver tumor status of Dab (Limanda limanda) from UK marine waters.
The flatfish species dab (Limanda limanda) is the sentinel for offshore marine monitoring in the United Kingdom National Marine Monitoring Programme (NMMP). At certain sites in the North and Irish Seas, the prevalence of macroscopic liver tumors can exceed 10%. The plasma proteome of these fish potentially contains reporter proteins or "biomarkers" that may enable development of diagnostic tests for liver cancer and further our understanding of the disease. Following selection of sample groups by quality-assured histopathology ("phenotype anchoring"), we used surface-enhanced laser desorption/ionization (SELDI) time-of-flight mass spectrometry to produce proteomic profiles of plasma from 213 dab collected during the 2004 UK NMMP. The resulting protein profiles were compared between fish from the North and Irish Seas and between fish with liver neoplasia or nondiseased liver. Significant differences were found between the plasma proteomes of dab from the North Sea and Irish Sea, which in conjunction with artificial neural networks can correctly determine from which sea dab were captured in 85% of the cases. In addition, the presence of liver tumors is associated with significant changes in the plasma proteome. We conclude that SELDI-based plasma profiling is potentially of use in nonlethal marine monitoring using wild sentinels such as dab. Furthermore, accurate selection of sample groups is critical for avoiding effects of confounding factors such as age, gender, and geographic origin of samples.


Medical history, lifestyle, and occupational risk factors for hairy cell leukemia: the InterLymph Non-Hodgkin Lymphoma Subtypes Project.
Little is known about the etiology of hairy cell leukemia (HCL), a rare B-cell lymphoproliferative disorder with marked male predominance. Our aim was to identify key risk factors for HCL. A pooled analysis of individual-level data for 154 histologically confirmed HCL cases and 8834 controls from five case-control studies, conducted in Europe and Australia, was undertaken. Age-, race and/or ethnicity-, sex-, and study-adjusted odds ratios (OR) and 95% confidence intervals (CI) were estimated using unconditional logistic regression. The usual patterns for age and sex in HCL were observed, with a median age of 55 years and sex ratio of 3.7 males to females. Cigarette smoking was inversely associated with HCL (OR = 0.51, 95% CI = 0.37 to 0.71) with dose-response relationships observed for duration, frequency, and lifetime cigarette smoking (P(trend) < .001). In contrast, occupation as a farmer was positively associated with HCL (OR = 2.34, 95% CI = 1.36 to 4.01), with a dose-response relationship observed for duration (OR = 1.82, 95% CI = 0.85 to 3.88 for ≤ 10 years vs never; and OR = 2.98, 95% CI = 1.50 to 5.93 for >10 years vs never; P(trend) = .025). Adult height was also positively associated with HCL (OR = 2.69, 95% CI = 1.39 to 5.29 for upper vs lower quartile of height). The observed associations remained consistent in multivariate analysis. Our observations of an increased risk of HCL from farming exposures and decreased risk from smoking exposures, independent of one another, support a multifactorial origin and an etiological specificity of HCL compared with other non-Hodgkin lymphoma subtypes. The positive association with height is a novel finding that needs replication.


Characterization of 2 human genotype G10 rotavirus strains, 3008CM and 1784/CI/1999, isolated in Cameroon and Cote d'Ivoire during the 1999-2000 rotavirus season.
During routine rotavirus surveillance projects in Cameroon and Cote d'Ivoire, 2 fecal samples collected from 2 children <5 years of age who presented with symptoms of gastroenteritis were found to give anomalous G typing results. These specimens were strongly rotavirus positive by enzyme immunoassay, displayed VP6 subgroup II specificity and long RNA electropherotypes, and were typed as rotavirus serotype G2 with monoclonal antibodies. In addition, the strains were typed as rotavirus VP7 genotype G3 and VP4 genotype P[8] by reverse-transcription polymerase chain reaction. Further investigation of the polymerase chain reaction G-typing results with a second set of primers revealed that the specimens were not genotype G3, and both samples were sequenced to elucidate the problem. Both strains were found to be genotype G10 by nucleotide sequence. Comparison of nucleotide and amino acid sequences and phylogenetic analysis of the African G10 strains revealed that these strains are closely related to the human G10 strains that were detected during the 2001-2003 rotavirus season in Ghana. The detection of G10 rotavirus in Africa adds to the global distribution of this strain and strengthens the need to continue strain surveillance in developing countries to understand the extent of strain distribution and diversity.


Prevalence of ocular morbidity among children aged 17 years or younger in the eastern India.
Childhood ocular morbidity involves a spectrum of eye diseases that critically impact the mental development, future education and quality of life. However, there is limited evidence about the early detection and appropriate treatment of ocular morbidity in children <20 years. This study was aimed to assess the prevalence and make a comparison between the different types of ocular morbidity in children of both sexes in the age group of 6-17 years in the eastern India. A cross-sectional survey of ocular morbidity among children <17 years of age who presented at the Department of Ophthalmology, Kalinga Institute of Medical Sciences, Bhubaneswar, and Vision Care Center for Retina, Bhubaneswar, in the eastern India between January 2015 and March 2018 was accomplished. Demographic information, visual acuity, type of eye injury, refractive errors and other detailed ophthalmic examination were screened. A total of 633 children (age 6-17 years) were examined in this study. The majority of cases were observed in children of age 12-17 years, accounting for almost close to half of all the cases. The prevalence of ocular morbidity was 45.92% in males and 53.97% in females. The most common ocular morbidity in children encountered was refractive error (54.62%), followed by congenital abnormalities (9%), allergic conjunctivitis (8.52%) and traumatic eye injury (7.1%). There was an increase in ocular morbidity with age, especially the refractive error and congenital abnormalities. A large number of ocular morbidity was observed in children of age <17 years. Since most of this morbidity was preventable or treatable, reasonable service for ocular morbidity and early age screening are effective methods to reduce this load. Moreover, health education for the prevention of childhood ocular morbidity and, at the same time, early presentation of children to ophthalmic hospitals for the treatment of eye disorders are essential.


Internalization of insulin receptors and HLA antigens in human hepatoma cells.
Human HepG2 hepatoma cells express a high number of insulin receptors. Growing cells exhibit 70% of their insulin receptors on the plasma membrane. Moreover, cell-surface insulin receptors form molecular complexes with class I major histocompatibility antigens, as determined by co-immunoprecipitation of the receptors by anti-class I monoclonal antibodies. On exposure to saturating concentrations of insulin, the hormone is rapidly internalized into a Pronase-resistant compartment. Internalization of insulin is accompanied by a rapid (t1/2 = 2-3 min) redistribution of insulin receptors from the cell surface to an intracellular compartment. On removal of insulin from the medium, functional receptors recycle back to the plasma membrane, where they can bind insulin again. With chronic exposure of HepG2 cells to insulin, the initial redistribution of receptors is followed by a slow (t1/2 = 9 h) down-regulation of the receptors. Finally, notwithstanding their interaction at the cell surface, insulin receptors and class I major histocompatibility antigens are internalized at different rates and with independent regulation.


Adaptive mgl-regulatory mutations and genetic diversity evolving in glucose-limited Escherichia coli populations.
The mutational adaptation of E. coli to low glucose concentrations was studied in chemostats over 280 generations of growth. All members of six independent populations acquired increased fitness through the acquisition of mutations at the mgl locus, increasing the binding protein-dependent transport of glucose. These mutations provided a strong fitness advantage (up to 10-fold increase in glucose affinity) and were present in most isolates after 140 generations. mgl constitutivity in some isolates was caused by base substitution, short duplication, small deletion and IS1 insertion in the 1041 bp gene encoding the repressor of the mgl system, mglD (galS). But an unexpectedly large proportion of mutations were located in the short mgl operator sequence (mglO), and the majority of mutations were in mglO after 280 generations of selection. The adaptive mglO substitutions in several independent populations were at exactly the positions conserved in the two 8 bp half-sites of the mgl operator, with the nature of the base changes also completely symmetrical. Either mutations were directed to the operator or the particular operator mutations had a selective advantage under glucose limitation. Indeed, isolates carrying mglO mutations showed greater rates of transport for glucose and galactose at low concentrations than those carrying mglD null mutations. mglO mutations avoid cross-talk by members of the GalR-Lacl repressor family, reducing transporter expression and providing a competitive advantage in a glucose-limited environment. Another interesting aspect of these results was that each adapted population acquired multiple mgl alleles, with several populations containing at least six different mgl-regulatory mutations co-existing after 200 generations. The diversity of mutations in the mglO/mglD region, generally in combination with mutations at other loci regulating glucose uptake (malT, mlc, ptsG), provided evidence for multiple clones in each population. Increased fitness was accompanied by the generation of genetic diversity and not the evolution of a single winner clone, as predicted by the periodic selection model of bacterial populations.


Effect of mono- and dichromatic light quality on growth rates and photosynthetic performance of Synechococcus sp. PCC 7002.
Synechococcus sp. PCC 7002 was grown to steady state in optically thin turbidostat cultures under conditions for which light quantity and quality was systematically varied by modulating the output of narrow-band LEDs. Cells were provided photons absorbed primarily by chlorophyll (680 nm) or phycocyanin (630 nm) as the organism was subjected to four distinct mono- and dichromatic regimes. During cultivation with dichromatic light, growth rates were generally proportional to the total incident irradiance at values <275 μmol photons m(-2) · s(-1) and were not affected by the ratio of 630:680 nm wavelengths. Notably, under monochromatic light conditions, cultures exhibited similar growth rates only when they were irradiated with 630 nm light; cultures irradiated with only 680 nm light grew at rates that were 60-70% of those under other light quality regimes at equivalent irradiances. The functionality of photosystem II and associated processes such as maximum rate of photosynthetic electron transport, rate of cyclic electron flow, and rate of dark respiration generally increased as a function of growth rate. Nonetheless, some of the photophysiological parameters measured here displayed distinct patterns with respect to growth rate of cultures adapted to a single wavelength including phycobiliprotein content, which increased under severely light-limited growth conditions. Additionally, the ratio of photosystem II to photosystem I increased ~40% over the range of growth rates, although cells grown with 680 nm light only had the highest ratios. These results suggest the presence of effective mechanisms which allow acclimation of Synechococcus sp. PCC 7002 acclimation to different irradiance conditions.


Endotoxin fever in granulocytopenic animals.
The febrile response to bacterial endotoxin was measured in rabbits made leukopenic with nitrogen mustard. A striking increase in susceptibility to the lethal effects of endotoxin occurred in severely leukopenic animals. Animals without circulating granulocytes, or with only basophils, developed no significant fever after endotoxin injection. Animals with circulating granulocytes other than basophils exhibited a biphasic febrile response to endotoxin; this response was significantly less in magnitude than that of control animals. Control animals, severely granulocytopenic animals, and animals with no circulating granulocytes other than basophils showed comparable febrile responses to serum pyrogen. These results suggest that granulocytes inactivate endotoxin in vivo and support the hypothesis that leukocyte pyrogen is a necessary intermediate in endotoxin fever. Basophils do not appear to participate in this process. These observations also contradict previous studies that were taken to indicate a normal febrile response to endotoxin in leukopenic animals and suggest that those results are related to the persistence of granulocytes.


Assessment of the swine protein-annotated oligonucleotide microarray.
The specificity and utility of the swine protein-annotated oligonucleotide microarray, or Pigoligoarray (http://www.pigoligoarray.org), has been evaluated by profiling the expression of transcripts from four porcine tissues. Tools for comparative analyses of expression on the Pigoligoarray were developed including HGNC identities and comparative mapping alignments with human orthologs. Hybridization results based on the Pigoligoarray's sets of control, perfect match (PM) and deliberate mismatch (MM) probes provide an important means of assessing non-specific hybridization. Simple descriptive diagnostic analyses of PM/MM probe sets are introduced in this paper as useful tools for detecting non-specific hybridization. Samples of RNA from liver, brain stem, longissimus dorsi muscle and uterine endothelium from four pigs were prepared and hybridized to the arrays. Of the total 20,400 oligonucleotides on the Pigoligoarray, 12,429 transcripts were putatively differentially expressed (DE). Analyses for tissue-specific expression [over-expressed in one tissue with respect to all the remaining three tissues (q < 0.01)] identified 958 DE transcripts in liver, 726 in muscle, 286 in uterine endothelium and 1027 in brain stem. These hybridization results were confirmed by quantitative PCR (QPCR) expression patterns for a subset of genes after affirming that cDNA and amplified antisense RNA (aRNA) exhibited similar QPCR results. Comparison to human ortholog expression confirmed the value of this array for experiments of both agricultural importance and for tests using pigs as a biomedical model for human disease.


A complex network representation of wind flows.
Climate networks have proven to be a valuable method to investigate spatial connectivity patterns of the climate system. However, so far such networks have mostly been applied to scalar observables. In this study, we propose a new method for constructing networks from atmospheric wind fields on two-dimensional isobaric surfaces. By connecting nodes along a spatial environment based on the local wind flow, we derive a network representation of the low-level circulation that captures its most important characteristics. In our approach, network links are placed according to a suitable statistical null model that takes into account the direction and magnitude of the flow. We compare a simulation-based (numerically costly) and a semi-analytical (numerically cheaper) approach to determine the statistical significance of possible connections, and find that both methods yield qualitatively similar results. As an application, we choose the regional climate system of South America and focus on the monsoon season in austral summer. Monsoon systems are generally characterized by substantial changes in the large-scale wind directions, and therefore provide ideal applications for the proposed wind networks. Based on these networks, we are able to reveal the key features of the low-level circulation of the South American Monsoon System, including the South American Low-Level Jet. Networks of the dry and the wet season are compared with each other and their differences are consistent with the literature on South American climate.


Benefits of supplementary tube feeding after fractured neck of femur: a randomised controlled trial.
A total of 744 elderly women with fractured neck of femur were classified into three groups according to anthropometric measurements on admission: group 1, well nourished; group 2, thin; group 3, very thin. Group 1 ate well and had a low mortality and a short rehabilitation time. The thinner the patients the lower their voluntary food intake, the higher their mortality and the longer their rehabilitation time. A series of 122 patients from groups 2 and 3 were entered postoperatively into a randomised controlled trial of overnight supplementary nasogastric tube feeding (4.2 MJ (1000 kcal), including 28 g protein) in addition to their normal ward diet. This treatment was associated with improvements not only in anthropometric and plasma protein measurements but also in clinical outcome, especially in the very thin group 3 patients. Rehabilitation time and hospital stay were shortened. Mortality in group 3 was less in the tube fed patients (8%) than in the controls (22%) but this difference did not reach statistical significance. One in five patients could not tolerate the nasogastric tube, but in the remainder the treatment caused no side effects and did not seriously diminish voluntary oral food intake by day.


Immunohistochemical status of the p53 protein and Ki-67 antigen using biopsied specimens can predict a sensitivity to neoadjuvant therapy in patients with esophageal cancer.
The prediction of an effective adjuvant therapy is essential in order to increase the number of esophageal cancer cases demonstrating an excellent response. Correlation of the immunohistochemical expression of p53 protein and Ki-67 antigen were studied using biopsied specimens of advanced esophageal cancer. The combined staining status of both can help predict the efficacy of neoadjuvant therapy in patients with esophageal cancer. The overexpression of p53 protein and the Ki-67 labeling percentage were immunohistochemically investigated in 95 biopsied specimens of advanced esophageal carcinoma resected between 1988 and 1996. All patients preoperatively received 1 course of either chemoradiotherapy or hyperthermo-chemoradiotherapy. Thirty-nine specimens were positive for p53 protein staining (41.1%), and the treatment modalities were histopathologically effective in 71.8% of these 39 patients (28/39), while the efficiency rate was 58.9% (33/56) in patients with p53(-). On the other hand, the efficiency rate of patients with a high Ki-67 labeling percentage (> or = 30%) was 73.9%, which was significantly higher than that of those with a low Ki-67 labeling percentage (< 30%) (38.5%, P = 0.0013). The efficacy rate of the patients with both a high Ki-67 labeling percentage and p53(+) was 80%, while that of the patients with a low Ki-67 labeling percentage and p53(-) was only 35.3% (P = 0.0098). The combination of a high Ki-67 labeling percentage and p53(-) particularly showed a high sensitivity to hyperthermo-chemoradiotherapy since the efficiency rate of these patients with hyperthermo-chemoradiotherapy was 81.5%, while the rate for those with chemoradiotherapy was 41.7% (P = 0.0129). An immunohistochemical analysis of p53 and the Ki-67 labeling percentage using biopsied specimens was thus found to effectively predict the efficacy of neoadjuvant therapies in patients with esophageal cancer.


Sprouty-4 inhibits transformed cell growth, migration and invasion, and epithelial-mesenchymal transition, and is regulated by Wnt7A through PPARgamma in non-small cell lung cancer.
Sprouty proteins are potent receptor tyrosine kinase inhibitors that antagonize growth factor signaling and are involved in lung development. However, little is known about the regulation or targets of Sprouty-4 (Spry4) in lung cancer. Our study aimed to determine the role of Spry4 in non-small cell lung cancer (NSCLC). We found that Spry4 mRNA expression was decreased in NSCLC cell lines and in dysplastic lung cell lines compared with a nontransformed cell line, suggesting that Spry4 has tumor-suppressing activity. When Spry4 was stably transfected into H157 and H2122 NSCLC cell lines, decreased migration and invasion were observed. Matrix metalloproteinase-9 activity was decreased, and the expression of matrix metalloproteinase inhibitors TIMP1 and CD82 were increased. Stable expression of Spry4 led to reduced cell growth and reduced anchorage-independent growth in NSCLC cell lines, along with upregulation of tumor suppressors p53 and p21. Changes in epithelial and mesenchymal markers indicated that Spry4 expression induces a reversal of the epithelial to mesenchymal transition characteristic of tumor cells. Treatment of a nontransformed lung epithelial cell line with short hairpin RNA to Spry4 led to the decreased expression of epithelial markers and increased cell growth, supporting the concept of Spry4 acting as a tumor suppressor. We showed that the activity of the Spry4 promoter is increased by Wnt7A/Fzd9 signaling through peroxisome proliferator-activated receptor gamma. These data present previously undescribed targets of Spry4 and suggest that Spry4 is a downstream target of Wnt7A/Fzd 9 signaling. Spry4 may have efficacy in the treatment of NSCLC.


Fate of arsenic in swine waste from concentrated animal feeding operations.
Swine diets are often supplemented by organoarsenicals, such as 3-nitro-4-hydroxyphenylarsonic acid (roxarsone) to treat animal diseases and promote growth. Recent work reported roxarsone degradation under anaerobic conditions in poultry litter, but no such data exist for swine wastes typically stored in lagoons nearby concentrated animal feeding operations (CAFOs). The objectives of this study were to: (i) characterize a suite of swine wastes collected from 19 randomly selected CAFOs for soluble arsenate [As(V)], arsenite [As(III)], dimethylarsenic acid (DMA), monomethylarsonic acid (MMA), 3-amino-4-hydroxyphenylarsonic acid (3-HPPA), p-arsanilic acid, and roxarsone, and (ii) determine the geochemical fate of roxarsone in storage lagoons nearby CAFOs. Swine waste suspensions were spiked with roxarsone and incubated under dark/light and aerobic/anaerobic conditions to monitor roxarsone degradation kinetics. Arsenic speciation analysis using liquid chromatography and inductively coupled plasma mass spectrometry (LC-ICPMS) illustrated the prevalence of As(V) in swine waste suspensions. Roxarsone underwent degradation to either organoarsenicals (3-HPPA) or As(V) and a number of unidentified metabolites. Roxarsone degradation occurred under anaerobic conditions for suspensions low in solids content, but suspensions higher in solids content facilitated roxarsone degradation under both anaerobic and aerobic conditions. Increased solids content enhanced roxarsone degradation kinetics under aerobic conditions. According to current waste storage and sampling practices, arsenic in swine wastes stored in lagoons has been overlooked as a possible environmental health issue.


Effects of hypoxia-cadmium interactions on rainbow trout (Oncorhynchus mykiss) mitochondrial bioenergetics: attenuation of hypoxia-induced proton leak by low doses of cadmium.
The goal of the present study was to elucidate the modulatory effects of cadmium (Cd) on hypoxia/reoxygenation-induced mitochondrial dysfunction in light of the limited understanding of the mechanisms of multiple stressor interactions in aquatic organisms. Rainbow trout (Oncorhynchus mykiss) liver mitochondria were isolated and energized with complex I substrates (malate-glutamate), and exposed to hypoxia (0>PO2<2 Torr) for 0-60 min followed by reoxygenation and measurement of coupled and uncoupled respiration and complex I enzyme activity. Thereafter, 5 min hypoxia was used to probe interactions with Cd (0-20 μmol l(-1)) and to test the hypothesis that deleterious effects of hypoxia/reoxygenation on mitochondria were mediated by reactive oxygen species (ROS). Hypoxia/reoxygenation inhibited state 3 and uncoupler-stimulated (state 3u) respiration while concomitantly stimulating states 4 and 4ol (proton leak) respiration, thus reducing phosphorylation and coupling efficiencies. Low doses of Cd (≤5 μmol l(-1)) reduced, while higher doses enhanced, hypoxia-stimulated proton leak. This was in contrast to the monotonic enhancement by Cd of hypoxia/reoxygenation-induced reductions of state 3 respiration, phosphorylation efficiency and coupling. Mitochondrial complex I activity was inhibited by hypoxia/reoxygenation, hence confirming the impairment of at least one component of the electron transport chain (ETC) in rainbow trout mitochondria. Similar to the effect on state 4 and proton leak, low doses of Cd partially reversed the hypoxia/reoxygenation-induced complex I activity inhibition. The ROS scavenger and sulfhydryl group donor N-acetylcysteine, administrated immediately prior to hypoxia exposure, reduced hypoxia/reoxygenation-stimulated proton leak without rescuing the inhibited state 3 respiration, suggesting that hypoxia/reoxygenation influences distinct aspects of mitochondria via different mechanisms. Our results indicate that hypoxia/reoxygenation impairs the ETC and sensitizes mitochondria to Cd via mechanisms that involve, at least in part, ROS. Moreover, we provide, for the first time in fish, evidence for a hormetic effect of Cd on mitochondrial bioenergetics--the attenuation of hypoxia/reoxygenation-stimulated proton leak and partial rescue of complex I inhibition by low Cd doses.


Synthetic Development of New 3-(4-Arylmethylamino)butyl-5-arylidene-rhodanines under Microwave Irradiation and Their Effects on Tumor Cell Lines and against Protein Kinases.
A new route to 3-(4-arylmethylamino)butyl-5-arylidene-2-thioxo-1,3-thiazolidine-4-one 9 was developed in six steps from commercial 1,4-diaminobutane 1 as starting material. The key step of this multi-step synthesis involved a solution phase "one-pot two-steps" approach assisted by microwave dielectric from N-(arylmethyl)butane-1,4-diamine hydrochloride 6a-f (as source of the first point diversity) and commercial bis-(carboxymethyl)-trithiocarbonate reagent 7 for construction of the rhodanine platform. This platform was immediately functionalized by Knoevenagel condensation under microwave irradiation with a series of aromatic aldehydes 3 as second point of diversity. These new compounds were prepared in moderate to good yields and the fourteen synthetic products 9a-n have been obtained with a Z-geometry about their exocyclic double bond. These new 5-arylidene rhodanines derivatives 9a-n were tested for their kinase inhibitory potencies against four protein kinases: Human cyclin-dependent kinase 5-p25, HsCDK5-p25; porcine Glycogen Synthase Kinase-3, GSK-3α/β; porcine Casein Kinase 1, SsCK1 and human HsHaspin. They have also been evaluated for their in vitro inhibition of cell proliferation (HuH7 D12, Caco 2, MDA-MB 231, HCT 116, PC3, NCI-H727, HaCat and fibroblasts). Among of all these compounds, 9j presented selective micromolar inhibition activity on SsCK1 and 9i exhibited antitumor activities in the HuH7 D12, MDA-MBD231 cell lines.


Primary subacute haematogenous osteomyelitis in children.
Between 1990 and 1998 we saw 21 children with primary subacute haematogenous osteomyelitis. Pain, swelling and a limp had been present for two to 12 weeks with little functional impairment. Laboratory tests were non-contributory. The lesions were classified radiologically into metaphyseal, diaphyseal, epiphyseal and vertebral. There were 24 sites involved, with most (20) being in the tibia; 17 lesions were in the diaphysis, five in the metaphysis and two in the epiphysis. The diagnosis was confirmed histologically in all cases. Staphylococcus aureus was cultured in six patients. Healing occurred in all patients after treatment with antibiotics for six weeks and radiological improvement was seen after three to six months. Subacute osteomyelitis develops as a result of increased host resistance and decreased bacterial virulence. The radiological features can mimic various benign or malignant bone tumours and non-pyogenic infections. Histological confirmation is necessary to avoid a delay in diagnosis.


In-hospital management of type 2 diabetes mellitus.
The increasing prevalence of type 2 diabetes brings with it a need to understand the particular impact of hospitalization in this patient population. Type 2 diabetes has been shown to increase length of stay, infection, and mortality rates. To optimize inpatient care, it is important to understand target glycemic goals as well as in-hospital glucose monitoring and diabetes management goals. A practical review of regimens for subcutaneous insulin administration,intravenous insulin infusion, and inpatient use of oral agents is presented. Methods for achieving adequate preparation and education of the patient and family for discharge to the outpatient setting are also discussed.


Utility of Quantitative EEG for Seizure Detection in Adults.
Traditional review of EEG for seizure detection requires time and the expertise of a trained neurophysiologist; therefore, it is time- and resource-intensive. Quantitative EEG (qEEG) encompasses a variety of methods to make EEG review more efficient and allows for nonexpert review. Literature supports that qEEG is commonly used by neurophysiologists and nonexperts in clinical practice. In this review, the different types of qEEG trends and spectrograms used for seizure detection in adults, from basic concepts to clinical applications, are discussed. The merits and drawbacks of the most common qEEG trends are detailed. The authors detail the retrospective literature on qEEG sensitivity, specificity, and false alarm rate as interpreted by experts and nonexperts alike. Finally, the authors discuss the future of qEEG as a useful screening tool and speculate on the trajectory of future investigations in the field.


Predictors of Smartphone and Tablet Use Among Patients With Hypertension: Secondary Analysis of Health Information National Trends Survey Data.
Uncontrolled hypertension leads to significant morbidity and mortality. The use of mobile health technology, such as smartphones, for remote blood pressure (BP) monitoring has improved BP control. An increase in BP control is more significant when patients can remotely communicate with their health care providers through technologies and receive feedback. Little is known about the predictors of remote BP monitoring among hypertensive populations. The objective of this study is to quantify the predictors of smartphone and tablet use in achieving health goals and communicating with health care providers via SMS text messaging among hypertensive patients in the United States. This study was a cross-sectional, secondary analysis of the 2017 and 2018 Health Information National Trends Survey 5, cycles 1 and 2 data. A total of 3045 respondents answered "Yes" to the question "Has a doctor or other healthcare provider ever told you that you had high blood pressure or hypertension?", which defined the subpopulation used in this study. We applied the Health Information National Trends Survey full sample weight to calculate the population estimates and 50 replicate weights to calculate the SEs of the estimates. We used design-adjusted descriptive statistics to describe the characteristics of respondents who are hypertensive based on relevant survey items. Design-adjusted multivariable logistic regression models were fitted to estimate predictors of achieving health goals with the help of smartphone or tablet and sending or receiving an SMS text message to or from a health care provider in the last 12 months. An estimated 36.9%, SE 0.9% (183,285,150/497,278,883) of the weighted adult population in the United States had hypertension. The mean age of the hypertensive population was 58.3 (SE 0.48) years. Electronic communication with the doctor or doctor's office through email or internet (odds ratio 2.93, 95% CI 1.85-4.63; P<.001) and having a wellness app (odds ratio 1.82, 95% CI 1.16-2.86; P=.02) were significant predictors of using SMS text message communication with a health care professional, adjusting for other demographic and technology-related variables. The odds of achieving health-related goals with the help of a tablet or smartphone declined significantly with older age (P<.001) and ownership of basic cellphones (P=.04). However, they increased significantly with being a woman (P=.045) or with being married (P=.03), having a wellness app (P<.001), using devices other than smartphones or tablets to monitor health (P=.008), making health treatment decisions (P=.048), and discussing with a provider (P=.02) with the help of a tablet or smartphone. Intervention measures accounting for age, gender, marital status, and the patient's technology-related health behaviors are required to increase smartphone and tablet use in self-care and SMS text message communication with health care providers.


The Role of General and Daily Control Beliefs for Affective Stressor-Reactivity Across Adulthood and Old Age.
General and situational control beliefs have been examined separately as buffers of the effects of daily stressors on affective well-being. However, general (trait) control beliefs reflect perceived ability to adapt, change, and influence overall life circumstances, whereas situational (daily) control beliefs reflect perceived ability to manage current circumstances and achieve desired outcomes. Using 9 weeks of daily reports from 150 adults aged 18-89 years, we examined the extent that general and daily control beliefs buffer the between- and within-person associations involving stressors and negative and positive affect (i.e., daily stress processes) and whether/how the extent of buffering differs with age. Aligning with prior findings, both greater average stressor exposure and experiencing a daily stressor compromised daily affective well-being and both higher general and daily control beliefs facilitated daily affective well-being. Specific to the motivating hypotheses, both general and daily control beliefs buffered daily stressor-reactivity. Age was associated with individuals' daily stressor-buffering, such that stressor-reactivity was more effectively damped at older ages. Associations between general control beliefs and daily stress processes were age invariant. Mixed evidence of age differences across general and daily control beliefs highlights how within-person processes may differentially contribute to well-being as individuals accommodate age-related strengths and vulnerabilities.


[Seasonal changes in cutaneous blood vessel tone in the frog Rana esculenta].
Experiments have been carried out on frogs immobilized by diplacine. The diameter of arteries and veins in the web was measured before and after sectioning the sciatic nerve. Denervation of the hindlimb resulted in dilatation of web vessels, which was different at various seasons. Changes in the arterial tone were found to be polyphasic. The highest tone was observed in March, April and September, the lowest one-in May, August, October and January. Changes in the venous tone are less pronounced and follow biphasic pattern, the tone being increased in spring and summer and decreased in autumn and winter. Stimulation of the peripheral end of the sectioned nerve leads to the constriction of web vessels, its amplitude being dependent on the level of vascular tone at the given period of year. It is suggested that seasonal changes in the tone of cutaneous arteries are partially based on changes in the reactivity of the blood vessels.


NF-kappaB blockade reduces the O2-evoked rise in Na+ conductance in fetal alveolar cells.
Electrophoretic mobility shift assays revealed minimal levels of NF-kappaB activity in rat distal lung epithelial cells cultured at fetal (23 mmHg) or adult alveolar (100 mmHg) P(O2), but revealed significant activation of this transcription factor in cells exposed to a rise in P(O2) mimicking that experienced at birth. This response was entirely abolished by pretreating cells with 5 mM sulfasalazine (SSA). This shift in P(O2) also evoked a rise in apical Na+ conductance (G(Na+)) that may underlie the O2-evoked stimulation of Na+ transport seen in these cells. Pretreatment with SSA had no effect upon G(Na+) in cells cultured continually at adult or fetal P(O2) but did inhibit the increase in G(Na+) seen in cells that had experienced the rise in P(O2). O2-evoked activation of NF-kappaB may thus mediate the increased Na+ transport that occurs when the distal lung epithelial cells are exposed to a physiologically-relevant increase in P(O2).


Diagnosing current distributions in batteries with magnetic resonance imaging.
Batteries and their defects are notoriously difficult to analyze non-destructively, and consequently, many defects and failures remain little noticed and characterized until they cause grave damage. The measurement of the current density distributions inside a battery could reveal information about deviations from ideal cell behavior, and could thus provide early signs of deterioration or failures. Here, we describe methodology for fast nondestructive assessment and visualization of the effects of current distributions inside Li-ion pouch cells. The technique, based on magnetic resonance imaging (MRI), allows measuring magnetic field maps during charging/discharging. Marked changes in the distributions are observed as a function of the state of charge, and also upon sustaining damage. In particular, it is shown that nonlinearities and asymmetries of current distributions could be mapped at different charge states. Furthermore, hotspots of current flow are also shown to correlate with hotspots in charge storage. This technique could potentially be of great utility in diagnosing the health of cells and their behavior under different charging or environmental conditions.


Expression of herpes simplex virus type 1 glycoprotein D deletion mutants in mammalian cells.
Glycoprotein D (gD) is a viron envelope component of herpes simplex virus types 1 and 2. We have previously defined seven monoclonal antibody (MAb) groups which recognize distinct epitopes on the mature gD-1 protein of 369 amino acids. MAb groups VII, II, and V recognize continuous epitopes at residues 11-19, 272-279, and 340-356, respectively. MAb groups I, III, IV, and VI recognize discontinuous epitopes. Recent studies have focused on epitopes I, III, and VI. Using truncated forms of gD generated by recombinant DNA methods and proteolysis, epitopes III, IV, and VI were located within amino acids 1-233. A portion of discontinuous epitope I was located in a region within residues 233-275. For this study, we used recombinant DNA methods to create mutations in the gD-1 gene and studied the effects of those mutations on gD as expressed in mammalian cells. Plasmid pRE4, containing the coding sequence of gD-1 and the Rous sarcoma virus long terminal repeat promoter, was transfected into mammalian cells. The expressed protein, gD-1-(pRE4), was identical in size and antigenic properties to gD-1 from infected cells. Six in-frame deletion mutations were subsequently constructed by using restriction enzymes to excise portions of the gD-1 gene. Plasmids carrying these mutated forms were transfected into cells, and the corresponding proteins were examined at 48 h posttransfection for antigenicity and glycosylation patterns. Three deletions of varying size were located downstream of residue 233. Analysis of these mutants showed that amino acids within the region 234-244 were critical for binding of DL11 (group I), but not for other MAb groups. Three other deletion mutants lost all ability to bind MAbs which recognize discontinuous epitopes. In addition, much of the gD expressed by these mutants was observed to migrate as high-molecular-weight aggregated forms in nondenaturing gels. Each of these mutations involved the loss of a cysteine residue, suggesting that disulfide linkages play an essential role in the formation of discontinuous epitopes. The extent of glycosylation of the mutant gD molecules accumulated at 48 h posttransfection suggested altered carbohydrate processing. In one case, there was evidence for increased O-linked glycosylation. Those proteins which had lost a cysteine residue as part of the deletion did not accumulate molecules processed beyond the high-mannose stage. The results suggest that carbohydrate processing during synthesis of gD is very sensitive to alterations in structure, particularly changes involving cysteine residues.


GnT-V expression and metastatic phenotypes in macrophage-melanoma fusion hybrids is down-regulated by 5-Aza-dC: evidence for methylation sensitive, extragenic regulation of GnT-V transcription.
Beta1,6-acetylglucosaminyltransferase V (GnT-V) forms beta1,6 branching on the trimannosyl terminus of N-glycans, allowing for the production of beta1,6Glc-NAc-bearing oligosaccharides. These are used by healthy myeloid cells and cancer cells alike for systemic migration. GnT-V has multiple glycoprotein substrates and thereby exerts global effects on cancer progression, characteristic of a master regulator of metastasis. Yet little is known of the regulation of GnT-V expression by tumor cells. It was previously reported that fusion of macrophages with Cloudman S91 mouse melanoma cells produced macrophage-melanoma hybrids with up-regulated GnT-V expression regarding mRNA and enzymatic activity. Majority of these hybrids showed increased chemotactic motility in vitro and elevated metastatic potential in vivo. Here we attempted to understand this at the molecular genetic level focusing on DNA hypermethylation as a potentially key step. Treatment of cells with 5-Aza-dC, an inhibitor of DNA methylation, resulted in decreased expression of GnT-V mRNA and beta1,6-branched oligosaccharides along with reduced glycosylation of LAMP-1, a major substrate for GnT-V. This was accompanied by reduced chemotactic motility of the cells. The results suggested that DNA hypermethylation in some fashion stimulated GnT-V expression. We thus investigated the promoter region of the GnT-V gene for hypermethylation of CpG islands, comparing macrophage-melanoma hybrids of low and high metastatic potential with the parental melanoma cell line. Genomic DNA after bisulfite modification amplified from this region showed identical sequences between the cell lines. The findings indicated that differential methylation of the promoter region of GnT-V gene was not responsible for its transcriptional control, rather, appeared to be controlled through a negative regulator, nm23, whose own expression was regulated by hypermethylation. Although our studies involved a highly experimental system, the results further suggest that by whatever mechanism, reduction of GnT-V activity through 5-Aza-dC treatment might provide a new approach towards prevention of metastatic progression.


Elevated Gastric Antrum Erosions in Portal Hypertension Patients: Peptic Disease or Mucosal Congestion?
Portal hypertension (PH) is a syndrome characterized by chronic increase in the pressure gradient between the portal vein and inferior vena cava. Previous studies have suggested an increased frequency of antral elevated erosive gastritis in patients with PH, as well as an etiologic association; however, there has not been any histological evidence of this hypothesis to date. Our aim was to evaluate the histological features found in elevated antral erosions in patients with portal hypertension. Sixty-nine patients were included; 28 with and 41 without PH. All patients underwent endoscopy, and areas with elevated antral erosion were biopsied. In the PH group, 24 patients had inflammatory infiltration with or without edema and vascular congestion, and 4 patients had no inflammation. In the group without PH, all patients showed inflammatory infiltration of variable intensity. There was no statistical significance between the two groups in the presence of Helicobacter pylori. There as a histological similarity between the two groups, if PH patients without inflammation were excluded; however, more edema and vascular congestion were observed in the PH group (p=0.002). The findings show that elevated antral erosions in patients with PH have more evident edema and vascular congestion in addition to lymphocytic infiltration.


Lactobacillus casei is able to survive and initiate protein synthesis during its transit in the digestive tract of human flora-associated mice.
Live Lactobacillus casei is present in fermented dairy products and has beneficial properties for human health. In the human digestive tract, the resident flora generally prevents the establishment of ingested lactic acid bacteria, the presence of which is therefore transient. The aim of this work was to determine if L. casei DN-114 001 survives during transit and how this bacterium behaves in the digestive environment. We used the human flora-associated (HFA) mouse model. L. casei DN-114 001 was genetically modified by the introduction of erm and lux genes, encoding erythromycin resistance and luciferase, respectively. For this modified strain (DN-240 041), light emission related to luciferase expression could easily be detected in the contents of the digestive tract. When inoculated into the digestive tract of HFA mice, L. casei (DN-240 041) survives but is eliminated with the same kinetics as an inert transit marker, indicating that it does not establish itself. In pure culture of L. casei, luciferase activities were high in the exponential and early stationary growth phases but decreased to become undetectable 1 day after inoculation. Viability was only slightly reduced even after more than 5 days. After transit in HFA mice, luciferase activity was detected even when 5-day-old L. casei cultures were given to the mice. In culture, the luciferase activity could be restored after 0.5 to 7 h of incubation in fresh medium or milk containing glucose, unless protein synthesis was inhibited by the addition of chloramphenicol or rifampin. These results suggest that in HFA mice L. casei DN-240 041, and thus probably L. casei DN-114 001, is able to initiate new protein synthesis during its transit with the diet. The beneficial properties of L. casei-fermented milk for human health might be related to this protein synthesis in the digestive tract.


Valproic acid enhances neuronal differentiation of sympathoadrenal progenitor cells.
The antiepileptic drug valproic acid (VPA) has been shown to influence the neural differentiation and neurite outgrowth of neural stem cells. Sympathoadrenal progenitor cells share properties with neural stem cells and are considered a potential cell source in the treatment of neurodegenerative diseases. The present study therefore aims at modulating the neural differentiation potential of these cells by treatment with the histone deacetylase inhibitor VPA. We studied the epigenetic effects of VPA in two culture conditions: suspension conditions aimed to expand adrenomedullary sympathoadrenal progenitors within free-floating chromospheres and adherent cell cultures optimized to derive neurons. Treatment of chromospheres with VPA may launch neuronal differentiation mechanisms and improve their neurogenic potential upon transplantation. However, also transplantation of differentiated functional neurons could be beneficial. Treating chromospheres for 7 days with clinically relevant concentrations of VPA (2 mm) revealed a decrease of neural progenitor markers Nestin, Notch2 and Sox10. Furthermore, VPA initiated catecholaminergic neuronal differentiation indicated by upregulation of the neuronal marker β-III-tubulin, the dopaminergic transcription factor Pitx3 and the catecholaminergic enzymes TH and GTPCH. In adherent neural differentiation conditions, VPA treatment improved the differentiation of sympathoadrenal progenitor cells into catecholaminergic neurons with significantly elevated levels of nor- and epinephrine. In conclusion, similar to neural stem cells, VPA launches differentiation mechanisms in sympathoadrenal progenitor cells that result in increased generation of functional neurons. Thus, data from this study will be relevant to the potential use of chromaffin progenitors in transplantation therapies of neurodegenerative diseases.


Cytoplasmic accumulation of the nuclear receptor CAR by a tetratricopeptide repeat protein in HepG2 cells.
The nuclear constitutive active receptor (CAR) is a key transcription factor regulating phenobarbital (PB)-inducible transcription of various hepatic genes that encode xenobiotic/steroid-metabolizing enzymes. CAR is retained in the cytoplasm of noninduced livers and translocates into the nucleus after PB induction. HepG2 cells lack the capability of retaining CAR in the cytoplasm; thus, the receptor spontaneously accumulates in the nucleus. We have now cloned and characterized a tetratricopeptide repeat (TPR) protein, designated cytoplasmic CAR retention protein (CCRP), for its ability to accumulate the receptor in the cytoplasm of cotransfected HepG2 cells. CCRP directly interacts with the ligand-binding domain of CAR and mediates the formation of a cytoplasmic CAR-CCRP-90-kDa heat shock protein (hsp90) ternary complex. Simultaneous expression of fluorescent protein-tagged CAR and CCRP reveals their colocalization with tubulin in mouse liver in vivo. Thus, these results indicate that CCRP may be a component of the CAR-hsp90 complex and involved in retaining the receptor in the cytoplasm of both HepG2 cells and probably in vivo liver cells.


[Anesthesia in breast feeding. Which restrictions are justified?].
In the last years knowledge of the excretion of drugs into breast milk has increased significantly. But information concerning anesthetic and analgesic drugs seems to be dispersed and limited. This paper describes the pharmacokinetic principles of drug transfer to milk and offers a comprehensive survey for drugs used in the perianesthetic period. The necessity of anesthesia should never be a sufficient reason to quit breast-feeding. With the information given in this article the individual duration of breast-milk-free interval can be discussed. Thus it will be possible to anesthetize a breast-feeding mother with minimal risks for the child without giving up the advantages of mother milk. Through careful selection of appropriate drugs even elective operations don't need to be postponed and breastfeeding can be continued as soon as in the immediate postoperative period.


Detection of antinuclear and antilaminin antibodies in autistic children who received thimerosal-containing vaccines.
Autism, a neurodevelopmental disorder, may involve autoimmune pathogenesis. Since mercury is potentially a risk factor for autoimmunity, we conducted a study of mercury-induced antinuclear and antilaminin antibodies in autistic and normal children who had been pre-administered with thimerosal-containing vaccines. Laboratory analysis by different immunoassays showed that the serum level of these two autoimmune markers did not significantly differ between autistic and normal children. This finding suggests that the mercury as in thimerosal-containing vaccines is likely not related to autoimmune phenomenon in autism.


Anti-idiotypic antibody vaccine for type B viral hepatitis in chimpanzees.
Anti-idiotypic antibodies (anti-Id) that contain an internal image component that mimics the surface antigen of hepatitis B virus (HBsAg) were used to immunize chimpanzees. Four injections of the rabbit anti-Id preparation elicited an antibody response to HBsAg (anti-HBs). The antibody specificity appeared to be against the anti-Id, since the anti-Id immunogen was shown to bind the chimpanzee anti-HBs. Two chimpanzees immunized with the anti-Id, along with two control animals that were either untreated or received a nonimmune rabbit immunoglobulin G preparation, were challenged with infectious hepatitis B virus. Both control chimpanzees developed clinical and serological characteristics consistent with an active hepatitis B virus infection, whereas the two anti-Id treated chimpanzees were protected from infection. Since chimpanzees provide a relevant model of a human response to hepatitis B virus immunization and infection, these results indicate that anti-Id preparations such as that described here might be candidates for vaccines against human diseases.


Outcome of extracranial malignant rhabdoid tumours in children registered in the European Paediatric Soft Tissue Sarcoma Study Group Non-Rhabdomyosarcoma Soft Tissue Sarcoma 2005 Study-EpSSG NRSTS 2005.
Extracranial malignant rhabdoid tumours (MRT) are rare lethal childhood cancers that often occur in infants and have a characteristic genetic mutation in the SMARCB1 gene. The European Paediatric Soft Tissue Sarcoma Study Group (EpSSG) conducted a multinational prospective study of registered cases of extracranial MRT to test an intensive multimodal approach of treatment for children with newly diagnosed extracranial MRT. Between December 2005 and June 2014, we prospectively registered 100 patients from 12 countries with a diagnosis of MRT tumour at an extracranial site on the EpSSG Non-Rhabdomyosarcoma Soft Tissue Sarcoma 2005 Study (NRSTS 2005). They were all treated on a standard multimodal protocol of surgery, radiotherapy, and chemotherapy over 30 weeks as follows: vincristine, cyclophosphamide, and doxorubicin (VDCy) at weeks 1, 10, 13, 22, and 28; vincristine was also given alone on weeks 2, 3, 11, 12, 14, 15, 23, 24, 29, and 30. Cyclophosphamide, carboplatin, and etoposide (Cy*CE) was given at weeks 4, 7, 16, 19, and 25. Radiotherapy was recommended for all primary tumour sites and all sites of metastatic disease. Forty-three patients completed the protocol treatment. Median follow-up for alive patients of the complete cohort was 44.6 months (range 11.5-84.6). For the whole cohort, the 3-year event-free survival (EFS) was 32.3% (95% confidence interval [CI] 23.2-41.6%) with a 3-year overall survival (OS) of 38.4% (95% CI 28.8-47.9%). For localised disease, the 4-year EFS was 39.3% (95% CI 28.2-50.1%) with a 4-year OS of 40.1% (95% CI 28.4-51.5%). For metastatic disease, the 2-year EFS was 8.7% (95% CI 1.5-24.2%) with a 2-year OS of 13.0% (95% CI 3.3-29.7%). Multivariable analysis disclosed that all patients ≤1 year of age were associated with at higher risk of death (hazard ratio [HR]: 2.6; 95% CI 1.0-6.8; p-value = 0.0094). Risk of death was also related with gender in metastatic patients (HR for males: 2.9, 95% CI 1.0-8.0; p-value = 0.0077). The EpSSG NRSTS 2005 protocol of intensive therapy can be delivered to extracranial MRT patients, with a possible improvement in outcome. The outcome, however, remains poor for patients who progress or with metastatic disease.


[Novel mutation in a patient with Carney complex].
Carney complex is a rare disease inherited in an autosomal dominant manner. It is mostly caused by inactivating mutations of the subunit of protein kinase A. Carney complex is associated with atrial myxoma, nevi or myxomas of the skin, breast tumors and endocrine overactivity. Primary pigmented nodular adrenocortical disease is the specific endocrine manifestation. The authors present the history of a 53-year-old female patient who had undergone surgery for atrial myxomas, thyroid tumor and breast cancer. She was also operated for an adrenal adenoma causing Cushing's syndrome. Genetic study revealed a mutation in the regulatory subunit of protein kinase A (ivs2-1G>A splice mutation in intron 2). Her heterozygous twins were also genetically screened and one of them carried the same mutation. The authors emphasize that despite the absence of specific treatment for patients with Carney complex, confirmation of the diagnosis by genetic studies is important for the close follow-up of the patient and early identification of novel manifestations.


[The effect of spermine on transport of Ca2+ ions in brain mitochondria].
The effect of spermine (50-400 microM) on the Ca-transporting system of brain mitochondria was studied. In a medium containing Mg2+ and ATP, spermine facilitates the accumulation of Ca2+ by decreasing Km of the uniporter. Spermine inhibits Na-stimulated Ca2+ efflux; this effect is dependent on the ionic strength of the medium--it is decreased when KCl concentration is increased from 20 to 120 mM. Spermine (200 microM) decreases (by 50%) the steady state concentration of Ca2+ maintained by mitochondria. The importance of spermine as a regulator of Ca2+-transport in brain mitochondria is discussed.


Synthesis of sterols by chick brain and liver during embryonic development.
The evolution throughout embryonic development of the rate at which acetate was converted into sterols was studied in chick brain and liver. Acetate incorporation (nmol/h/g tissue) was clearly higher in brain than in liver and sharply decreased with the age of embryo. Cholesterol and desmosterol were the major sterols formed from acetate by chick embryo brain, followed by lanosterol and squalene. No desmosterol was found in chick embryo liver, organ where cholesterol was the major sterol synthesized. In brain, the relative percentage of cholesterol increased throughout embryonic development reaching more than 50% at hatching, while the percentage of desmosterol decreased during the same period and represented at hatching only about 10-15% of the total nonsaponifiable fraction. The relative percentages of lanosterol and squalene did not change significantly throughout the period assayed. In liver, the percentage of cholesterol increased until 19 days but sharply decreased at hatching.


Laser surface structuring of diamond with ultrashort Bessel beams.
We investigate the effect of ultrafast laser surface machining on a monocrystalline synthetic diamond sample by means of pulsed Bessel beams. We discuss the differences of the trench-like microstructures generated in various experimental conditions, by varying the beam cone angle, the energy and pulse duration, and we present a brief comparison of the results with those obtained with the same technique on a sapphire sample. In diamond, we obtain V-shaped trenches whose surface width varies with the cone angle, and which are featured by micrometer sized channels having depths in the range of 10-20 μm. By laser writing crossed trenches we are also able to create and tailor on the diamond surface pillar-like or tip-like microstructures potentially interesting for large surface functionalization, cells capturing and biosensing.


Reexamination of the mechanism of hydroxyl radical adducts formed from the reaction between familial amyotrophic lateral sclerosis-associated Cu,Zn superoxide dismutase mutants and H2O2.
Amyotrophic lateral sclerosis (ALS) involves the progressive degeneration of motor neurons in the spinal cord and motor cortex. Mutations to Cu,Zn superoxide dismutase (SOD) linked with familial ALS are reported to increase hydroxyl radical adduct formation from hydrogen peroxide as measured by spin trapping with 5, 5'-dimethyl-1-pyrrolline N-oxide (DMPO). In the present study, we have used oxygen-17-enriched water and H2O2 to reinvestigate the mechanism of DMPO/.OH formation from the SOD and SOD mutants. The relative ratios of DMPO/.17OH and DMPO/.16OH formed in the Fenton reaction were 90% and 10%, respectively, reflecting the ratios of H217O2 to H216O2. The reaction of the WT SOD with H217O2 in bicarbonate/CO2 buffer yielded 63% DMPO/.17OH and 37% DMPO/.16OH. Similar results were obtained from the reaction between familial ALS SOD mutants and H217O2: DMPO/.17OH (64%); DMPO/.16OH (36%) from A4V and DMPO/.17OH (62%); and DMPO/.16OH (38%) from G93A. These results were confirmed further by using 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide spin trap, a phosphorylated analog of DMPO. Contrary to earlier reports, the present results indicate that a significant fraction of DMPO/.OH formed during the reaction of SOD and familial ALS SOD mutants with H2O2 is derived from the incorporation of oxygen from water due to oxidation of DMPO to DMPO/.OH presumably via DMPO radical cation. No differences were detected between WT and mutant SODs, neither in the concentration of DMPO/.OH or DEPMPO/.OH formed nor in the relative incorporation of oxygen from H2O2 or water.