[Comparative analysis of the use of levamisole and pyrogenal in the treatment of multiple sclerosis].
A series of 120 patients with multiple sclerosis was subjected to a clinico-immunological examination and treatment. In a group of patients receiving levamisole, improvement of the neurologic symptomatology was observed in 65% of the cases and that of the immunological indices in 97.5%. Pyrogenal treatment led to no improvement in the immunological parameters. Clinical improvement in this group was seen in only 17.8% of the patients. It is recommended that treatment with levamisole be conducted following the preliminary assessment of the sensitivity of the patient's lymphocytes to the drug in vitro.


A simplified pocket reduction procedure around a stud attachment without suturing.
Plaque accumulation on the implant titanium surface plays a critical role in the initiation and progression of periimplant disease. Removing plaque and calculus deposits, and reducing the periimplant probing depth, therefore, are important. The reduction of the periimplant probing depth in periimplant disease by using a cord and a surgical dressing pack has not been previously reported. This procedure is useful for periimplant mucositis and mild periimplantitis before a surgical intervention and can maintain a narrow keratinized mucosa with less pain than surgical intervention.


Aggressive hepatocellular carcinoma complicating pregnancy after autologous bone marrow transplantation for non-Hodgkin's lymphoma.
In the Asia-Pacific region, autologous and allogeneic bone marrow transplantation (BMT) in patients infected with the hepatitis B virus (HBV) may be complicated by fatal hepatic failure due to viral reactivation. Survivors may suffer from accelerated hepatitis and cirrhosis. We report the first case of hepatocellular carcinoma (HCC) after autologous BMT for mediastinal B cell lymphoma. The tumor developed rampantly during a planned pregnancy 5 years after BMT. Less than 40 cases of HCC complicating pregnancy have been reported, and outcome is invariably poor. Immunosuppression and HBV reactivation after autologous BMT, as well as immune tolerance and hormonal changes associated with pregnancy may contribute to the rapid tumor growth. Biochemical and radiological surveillance for HCC should be strengthened in HBV carriers after BMT, especially in patients with the histology of chronic liver disease, or biochemical/ virological evidence of increased HBV activity.


Downregulation of miR-15a due to LMP1 promotes cell proliferation and predicts poor prognosis in nasal NK/T-cell lymphoma.
Nasal NK/T-cell lymphoma (NNKTL) is an Epstein-Barr virus (EBV)-associated malignancy and has distinct clinical and histological features. However, its genetic features are hitherto unclear. MicroRNAs (miRNAs) play a crucial role in the pathogenesis of several malignancies via regulating gene expression. In this study, we investigated whether the specific microRNAs were related to the tumor behaviors in NNKTL. MiRNA array and Quantitative RT-PCR analyses revealed that miR-15a was expressed at a much lower level in NNKTL cells (SNK-1, SNK-6, and SNT-8) than in normal peripheral NK cells and EBV-negative NK cell line KHYG-1. Quantitative PCR and western blot analyses showed that the expression of MYB and cyclin D1, which are validated targets of miR-15a, was higher in NNKTL cells. Transfection of NNKTL cells (SNK-6 and SNT-8) with a miR-15a precursor decreased MYB and cyclin D1 levels, thereby blocking G1/S transition and cell proliferation. Knockdown of EBV-encoded latent membrane protein 1 (LMP1) significantly increased miR-15a expression in SNK-6 cells. In NNKTL tissues, we found that reduced miR-15a expression, which correlated with MYB and cyclin D1 expression, was associated with poor prognosis of NNKTL patients. These data suggest that downregulation of miR-15a, possibly due to LMP1, implicates in the pathogenesis of NNKTL by inducing cell proliferation via MYB and cyclin D1. Thus, miR-15a could be a potential target for antitumor therapy and a prognostic predictor for NNKTL.


The CRH1 antagonist CP154,526 failed to alter ischemia-induced neurodegeneration and spatial memory deficits in rats but inhibited behavioral activity in the novel open field.
Corticotropin-releasing hormone (CRH) has been implicated in ischemia-induced neurotoxicity, due in part to excitatory effects at the hippocampus, and the demonstrated neuroprotective effects of centrally administered, non-specific CRH antagonists. However, a number of issues remain to be clarified from these studies, including the relative contribution of CRH receptor subtypes, and the efficacy of these compounds to alter ischemia-induced behavioral impairments. In the current study, a highly selective, systemically administered CRH1 antagonist (CP154,526) failed to reverse global ischemia-induced cell death in hippocampal CA1 neurons or spatial memory impairments as assessed in the radial arm maze. Similarly, central administration of alpha-helical CRH failed to confer protection against ischemic damage. Interestingly, CRH1 antagonism reversed ischemia-induced hyperactivity in a novel open field, suggesting that modulation of this behavior is independent of effects on hippocampal CA1 cell loss. Failure of the current study to demonstrate neuroprotective effects of either the selective or non-selective CRH antagonists tested challenges the proposed neurotoxic role of CRH in global ischemia. These findings are discussed in relationship to recent findings reconsidering the participation of CRH in excitotoxic-mediated cellular damage.


Recent technological strategies for enhancing the stability of lycopene in processing and production.
Lycopene, a popular antioxidant, exists abundantly in nature in all-E form, but the Z-lycopene has better solubility and absorption characteristics. Lycopene is labile to light, thermal and oxygen. Emerging technological approaches are developed to improve the stability of lycopene during production and processing. This review presents the degradation mechanism of lycopene. A comprehensive evaluation has made by comparing the advantages and disadvantages of different extraction and isomerization methods. Encapsulation of lycopene in different delivery systems can further improve its stability. The process and formulation influence factors, as well as the advantages and disadvantages of lycopene encapsulation delivery systems are summarized. In the future, it is necessary to scientifically design low-cost and high-efficiency lycopene production methods.The stable and high bioavailable lycopene encapsulation systems need to be designed. Further study on the chemical stability mechanism of lycopene, the toxicity of biological materials, the safety of lycopene preparations also need to be evaluated.


Health risks from radioactive particles on Cumbrian beaches near the Sellafield nuclear site.
A monitoring programme, in place since 2006, continues to recover radioactive particles (<2 mm diameter) and larger objects from the beaches of West Cumbria. The potential risks to members of the public using the beaches are mainly related to prolonged skin contact with or the inadvertent ingestion of small particles. Most particles are classified as either 'beta-rich' or 'alpha-rich' and are detected as a result of their caesium-137 or americium-241 content. Beta-rich particles generally also contain strontium-90, with90Sr:137Cs ratios of up to about 1:1, but typically <0.1:1. Alpha-rich particles contain plutonium isotopes, with Pu:241Amαratios usually around 0.5-0.6:1. 'Beta-rich' particles have the greatest potential to cause localised skin damage if held in stationary contact with the skin for prolonged periods. However, it is concluded that only particles of >106Bq of137Cs, with high90Sr:137Cs ratios, would pose a significant risk of causing acute skin ulceration. No particles of this level of activity have been found. Inadvertent ingestion of a particle will result in the absorption to blood of a small proportion of the radionuclide content of the particle. The subsequent retention of radionuclides in body organs and tissues presents a potential risk of the development of cancer. For 'beta-rich' particles with typical activities (mean 2 × 104Bq137Cs, Sr:Cs ratio of 0.1:1), the estimated committed effective doses are about 30µSv for adults and about 40µSv for 1 year old infants, with lower values for 'alpha-rich' particles of typical activities. The corresponding estimates of lifetime cancer incidence following ingestion for both particle types are of the order of 10-6for adults and up to 10-5for infants. These estimates are subject to substantial uncertainties but provide an indication of the low risks to members of the public.


Outlining the nascent nervous system of Branchiostoma floridae (amphioxus) by the pan-neural marker AmphiElav.
The cephalochordate amphioxus is the closest living relative of the vertebrates. The anatomy and development of the amphioxus nervous system is giving insights into the basic chordate neural organisation, prior to the increase in neural complexity that characterised the origin of vertebrates. We have analysed the expression of the pan-neural marker AmphiElav in the Florida lancelet (Branchiostoma floridae) through development and up to larval stages. AmphiElav is expressed in the developing neural tube with particular temporal and spatial dynamics that correlate with neuronal differentiation. In addition, AmphiElav is expressed in isolated epidermal cells from mid-neurula embryos. This epidermal expression probably corresponds to precursors of sensory neurons. Together with data from other neural markers, we discuss the evolutionary relevance of these neural precursors to the origin of some of the most dramatic vertebrate inventions, neural crests and placodes.


Effects of long-term exposure to particulate matter and metal components on mortality in the Rome longitudinal study.
The effect of long-term exposure to metal components in particulate matter on mortality are still controversial. To study the association between long-term exposure to PM10, PM2.5, PM2.5 absorbance, particulate matter components (copper, iron, zinc, sulfur, silicon, potassium, nickel, and vanadium) and non-accidental, cardiovascular (CVD), and ischemic heart disease (IHD) mortality. All 30+ year olds from the Rome Longitudinal Study were followed for vital status from October 2001 until December 2010. We used land use regression models to estimate annual average concentrations at residences and Cox models to estimate the associations between pollutants and cause-specific mortality, adjusting for individual and contextual characteristics. Hazard ratios (HRs) were expressed per increments equal to the 5th-95th percentile range of each pollutant distribution. We analyzed 1,249,108 residents and found strong associations between all exposure indicators and mortality. We observed higher mortality risk with increasing exposure to PM2.5 absorbance (HR=1.05; 95% CI: 1.03-1.06) and to tracers of non-tailpipe traffic emissions such as tire and brake wear (Cu, Fe, and Zn); for PM2.5Zn, we found HR=1.06 (95% CI: 1.04-1.08) for non-accidental mortality, HR=1.07 (95% CI: 1.04-1.10) for CVD, and HR=1.11 (95% CI: 1.06-1.16) for IHD mortality. With increasing levels of nickel in PM10, we found HR=1.07 (95% CI: 1.05-1.09) for non-accidental mortality, HR=1.08 (95% CI: 1.05-1.11) for CVD, and HR=1.13 (95% CI: 1.08-1.18) for IHD mortality. Results were robust when we adjusted for PM mass and for cardiovascular mortality when we adjusted for NO2. In addition to vehicular exhaust pollutants, PM related to non-tailpipe emissions and mixed oil burning/industry plays an important role in mortality.


Intake of DDT and its metabolites through food items among reproductive age women in Bangladesh.
This study was conducted to make clear the major intake route of DDT in Bangladesh people to develop strategy and policy that could lead to a reduction in body burden especially in the reproductive age women. The concentrations of several POPs (DDT, PCBs, chlordanes, HCHs, HCB, and PeCB) were quantified in food items, human breast milk and house dust collected in Bangladesh in 2011-2012. Among the POPs analyzed in this study, DDT and its metabolites (ΣDDT) showed the highest concentration. The highest median ΣDDT concentration was found in meat (1.3-1100 ng g-1 wet weight) and house dust (30-1100 ng g-1 dry weight), and followed by human breast milk (20-55 ng g-1 wet weight). Estimated daily intake (EDI) was calculated using the DDT concentrations in food items. The highest intake of DDT was found in an infant and 99% of it was via breast milk feeding. DDT intake via consumption of beef accounted for 69% and 72% of that found in children and adults, respectively. The total EDI of DDT did not exceed the tolerable daily intake proposed by the WHO, but the EDI of p,p'-DDT exceeded the oral reference dose proposed by the US Environmental Protection Agency. Further research is required to clarify the reason for the high levels of DDT in beef, which seems to be the major intake route of DDT for women of reproductive age in Bangladesh.


Pathogenesis of lupus-like nephritis through autoimmune antibody produced by CD180-negative B lymphocytes in NZBWF1 mouse.
Toll-like receptors appear to play an important role in the pathogenesis of lupus-like nephritis in mice. In human and mouse, CD180 is a homologue of TLR4. In SLE patients, the number of CD180-negative B cells in peripheral blood changes in parallel with disease activity. In the present study using NZBWF1 mice, the population of splenic CD180-negative B cells increased with progression of renal lesions and aging. These cells produced both anti-dsDNA and histone antibodies; the peripheral blood levels of anti-dsDNA antibody increased markedly with aging. B cells infiltrating into renal lesions were CD180-negative and produced anti-dsDNA antibody. Considered together, these findings indicate that CD180-negative B cells contribute significantly to development of SLE-like morbidity in NZBWF1 mice by autoantibody production.


Differential expression of regulator of G-protein signaling R12 subfamily members during mouse development.
Regulators of G-protein Signaling (RGS proteins) are a multigene family of GTPase-accelerating proteins for the Galpha subunit of heterotrimeric G-proteins. The mammalian R12 RGS protein subfamily is composed of RGS12 and RGS14, two proteins characterized by their multidomain architecture of hallmark RGS domain, tandem Ras-binding domains (RBDs), and a second Galpha interacting domain, the GoLoco motif. The Rgs12 gene generates multiple splice variants, the largest of which encodes N-terminal PDZ and PTB domains in addition to the core RGS/RBD/GoLoco motifs. The Rgs14 gene encodes a protein similar to the non-PDZ/PTB domain RGS12 splice variants. The spatiotemporal expression patterns of RGS12 and RGS14 proteins were examined by immunohistochemistry in a developmental series of postimplantation mouse embryo. We report that RGS12 splice variants exhibit differential spatiotemporal patterns of expression during postimplantation embryogenesis, suggesting nonoverlapping roles. In contrast, RGS14 is found ubiquitously throughout the postimplantation period. We conclude that R12 subfamily RGS proteins likely play significant and different roles in specific tissues and periods of mouse embryogenesis.


Distribution of chondrocytes containing alpha-smooth muscle actin in human normal, osteoarthrotic, and transplanted articular cartilage.
The aim of our study was to evaluate the occurrence of chondrocytes containing alpha-smooth muscle actin in human normal and diseased cartilage. Immunohistochemistry using monoclonal antibodies for alpha-smooth actin, muscle-specific actin, S-100 protein, CD 34, and desmin was performed on samples of human articular cartilage obtained at autopsy following sudden death, during total hip and knee replacement for osteoarthritis, or after femoral neck fracture in patients without symptoms of osteoarthritis. Moreover, the layers of residual cartilage from chondral posttraumatic defects obtained during preoperative arthroscopy and of newly formed cartilage after autologous-chondrocyte transplantation (Hyalograft C) obtained during second-look arthroscopy were also examined by immunohistochemistry and RT PCR. Our study showed that a significant percentage of articular chondrocytes express alpha-smooth muscle actin in healthy, diseased, and regenerated articular cartilage. Alpha-actin positive chondrocytes (18%) were observed predominantly in the upper zone of normal articular cartilage. By contrast, only approximately 10% of cartilage cells in the deep region stained for this contractile actin isoform. Actin-positive chondrocytes (myochondrocytes) are formed predominantly in response to injury to the osteoarthrotic cartilage, at sites of defective healing, and in newly formed cartilage after autologous chondrocyte transplantation. Fibrocartilage is present in some of these conditions, and it is known that this tissue contains chondrocytes with actin. The presence of myochondrocytes in the surface layer of normal articular cartilage indicates that this region probably plays an important role in maintaining cartilage integrity. Myochondrocytes may utilize the contractile actin isoform in manipulating the extracellular matrix of articular cartilage. It is also possible that actin-containing chondrocytes have a higher potential for regeneration in contrast to chondrocytes that do not contain this contractile material in their cytoplasm.


Primary structure of human alpha 2-macroglobulin. IV. Primary structure of two large CNBr fragments, located in the COOH-terminal part and accounting for 337 residues.
The amino acid sequences have been determined for two CNBr fragments of human alpha 2-macroglobulin which, due to the presence of an uncleaved Hse-Thr bond, form an Mr = 40,000 fragment. These fragments are located in the COOH-terminal part of alpha 2-macroglobulin (CB21, residues 955-1185 and CB22, residues 1186-1291). CB21 contains one glucosamine-based carbohydrate group attached to Asn-14 and one internal disulfide bridge (Cys-102 bound to Cys-150). CB21 and CB22 account for 337 of the 1451 residues of the subunit of alpha 2-macroglobulin.


Unfavourable outcomes of drug therapy--subjective probability versus confidence intervals.
To investigate how individual predictions compare with confidence intervals, we asked 50 medical residents and 28 graduate students with biostatistics training to estimate unfavourable outcomes of drug therapy (therapeutic failures and side-effects) in groups of 10 and 20 patients. The predictions made by physicians and graduate students were similar for both sample sizes and types of outcome. The majority (58%) of estimates were greater than the upper limit of the 95% confidence interval, a bias that may hamper the correct interpretation of therapeutic risks in medical decision-making.


Development and validation of core entrustable professional activities for abdominal radiology.
To develop and validate European entrustable professional activities (EPAs) for sub-specialised hepatobiliary and gastrointestinal (HB/GI) diagnostic imaging. Both European Society of Radiology and national curricula in HB/GI diagnostic radiology were thoroughly reviewed, resulting in preliminary EPAs drafted by a pilot group of expert radiologists in 2 different countries. Each EPA was fully described with 7 components (Specification/limitations; Potential risks of failing; Relevant domains of competence; Required experience, knowledge, skills, attitude and behaviour; Assessment information sources to assess progress and ground a summative entrustment decision; Entrustment for which level of supervision is to be reached; and Expiration date). The modified Delphi method with 3 Delphi rounds was chosen for validation. Content validity index (CVI) and median values were used for validation. There were 15 preliminary EPAs, some of them divided according to 2 levels: resident and fellow level. The 37 members of the Delphi group were based in 2 different European countries with a background experience of 10 represented countries. Subsequent to the first Delphi round, 6 EPAs were accepted (CVI ≥ 0.8, median ≥ 4), 6 needed major revisions (CVI 0.7-0.79, median ≥ 4), 3 were rejected (CVI < 0.7) and 1 was added. After the second Delphi round, both the 6 revised EPAs and the additional one met the validation criteria (CVI ≥ 0.8, median ≥ 4). Finally, 13 EPAs were validated during the 3rd Delphi round with an agreement percentage of 95-100%. This study creates and validates EPAs for sub-specialised HB/GI diagnostic imaging. Thirteen EPAs for sub-specialised hepatobiliary and gastrointestinal diagnostic imaging were created with a strong methodology, and as a first example set in sub-specialised diagnostic imaging, they provide a template for others to be created. • The competence-based teaching in medical studies has recently been reintroduced through EPAs. • Thirteen EPAs have been developed for hepatobiliary and gastrointestinal sub-specialised diagnostic imaging. • These EPAs were validated using a Delphi modified method and provide a template for other to be created.


Proteomic cancer classification with mass spectrometry data.
The ultimate goal of cancer proteomics is to adapt proteomic technologies for routine use in clinical laboratories for the purpose of diagnostic and prognostic classification of disease states, as well as in evaluating drug toxicity and efficacy. Analysis of tumor-specific proteomic profiles may also allow better understanding of tumor development and the identification of novel targets for cancer therapy. The biological variability among patient samples as well as the huge dynamic range of biomarker concentrations are currently the main challenges facing efforts to deduce diagnostic patterns that are unique to specific disease states. While several strategies exist to address this problem, we focus here on cancer classification using mass spectrometry (MS) for proteomic profiling and biomarker identification. Recent advances in MS technology are starting to enable high-throughput profiling of the protein content of complex samples. For cancer classification, the protein samples from cancer patients and noncancer patients or from different cancer stages are analyzed through MS instruments and the MS patterns are used to build a diagnostic classifier. To illustrate the importance of feature selection in cancer classification, we present a method based on support vector machine-recursive feature elimination (SVM-RFE), demonstrated on two cancer datasets from ovarian and lung cancer.


Ferritin and iron uptake by reticulocytes.
The uptake of liver ferritin labelled with 125I or 59Fe by guinea-pig reticulocytes has been studied to investigate the characteristics of the uptake process, compare it with transferrin uptake and determine whether ferritin-iron is utilized by the cells in haem synthesis. The results confirmed that guinea-pig reticulocytes, but not mature erythrocytes, take up liver ferritin by a saturable, time- and temperature-dependent process. Up to 70% of the iron taken up by the cells was utilized in haem synthesis and competed directly with iron derived from transferrin. Scatchard analysis of the binding parameters indicated that 30-130 X 10(3) ferritin molecules were bound per cell to high affinity specific membrane receptors (Ka: 1.77 X 10(7) M-1). In contrast, rat took up much less ferritin than guinea-pig reticulocytes and the process was entirely non-specific. Release experiments with guinea-pig reticulocytes at 37 degrees C showed that a maximum of about 70% of the cell-associated 125I-ferritin was released from the cells of which up to 15% was trichloroacetic acid-soluble. We suggest that ferritin uptake by guinea-pig reticulocytes involves receptor-mediated endocytosis. The endocytotic vesicle fuses with a lysosome, iron is removed from the protein and enters a cytosolic pool in which it competes directly with transferrin-derived iron to provide iron for mitochondrial haem synthesis. Some of the ferritin is catabolized and the rest is returned to the extracellular medium during membrane recycling.


A novel thermophilic hemoprotein scaffold for rational design of biocatalysts.
Hemoproteins are commonly found in nature, and involved in many important cellular processes such as oxygen transport, electron transfer, and catalysis. Rational design of hemoproteins can not only inspire novel biocatalysts but will also lead to a better understanding of structure-function relationships in native hemoproteins. Here, the heme nitric oxide/oxygen-binding protein from Caldanaerobacter subterraneus subsp. tengcongensis (TtH-NOX) is used as a novel scaffold for oxidation biocatalyst design. We show that signaling protein TtH-NOX can be reengineered to catalyze H2O2 decomposition and oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) by H2O2. In addition, the role of the distal tyrosine (Tyr140) in catalysis is investigated. The mutation of Tyr140 to alanine hinders the catalysis of the oxidation reactions. On the other hand, the mutation of Tyr140 to histidine, which is commonly observed in peroxidases, leads to a significant increase of the catalytic activity. Taken together, these results show that, while the distal histidine plays an important role in hemoprotein reactions with H2O2, it is not always essential for oxidation activity. We show that TtH-NOX protein can be used as an alternative scaffold for the design of novel biocatalysts with desired reactivity or functionality. H-NOX proteins are homologous to the nitric oxide sensor soluble guanylate cyclase. Here, we show that the gas sensor protein TtH-NOX shows limited capacity for catalysis of redox reactions and it can be used as a novel scaffold in biocatalysis design.


Determination of serum levels of YKL-40 and hyaluronic acid in patients with hepatic fibrosis due to schistosomiasis japonica and appraisal of their clinical value.
The objectives of this study were (i) to evaluate the serum levels of human cartilage glycoprotein 39 (YKL-40) and hyaluronic acid (HA) in people infected with Schistosoma japonicum, and (ii) to determine their clinical value. A total of 563 people were subjected to ultrasonography, and 60 patients were identified with either mild (n=30) or severe (n=30) hepatic fibrosis. In addition, 28 healthy subjects were included as controls. Blood sera of these 88 people were examined with regard to the levels of YKL-40 and HA. The former was measured by enzyme-linked immunosorbent assay, and serum HA was determined by a commercially available radioimmunoassay method. On the basis of the ultrasonographic investigations, HA levels in normal, mild, and severe cases of hepatic fibrosis were 83.0+/-35.7, 216.1+/-77.9 and 212.6+/-80.9 microg/ml, respectively. When the same sera were tested for YKL-40, 49.0+/-10.4, 92.3+/-18.5 and 172.1+/-35.9 microg/ml, respectively, were recorded in the three groups. Thus, the serum levels of YKL-40 are not only increased in patients infected with S. japonicum but they are also correlated with the stage of hepatic fibrosis. In conclusion, it appears that YKL-40 is more sensitive than HA in measuring the degree of hepatic fibrosis due to schistosomiasis, which warrants further investigation.


Quantitative cerebral water content mapping in hepatic encephalopathy.
There is increasing evidence that the pathophysiology of hepatic encephalopathy is tightly associated with low-grade cerebral oedema; however, no method has yet specifically and unambiguously confirmed this hypothesis in vivo. The current study describes the quantitative measurement of localised water content using MRI in a cohort of 38 patients suffering from hepatic encephalopathy. A significant global increase in cerebral water content was observed in white matter whereas water content in grey matter was globally unaffected. However, significant spatial variations in the water content distribution, especially in grey matter, were observed and were correlated with disease grade and critical flicker frequency. In addition, regions-of-interest were defined and a significant change in water content with disease grade was found in the frontal and occipital white matter, the globus pallidus, the anterior limb of the internal capsule and the putamen. No association of water content and HE grade was established for the occipital visual and frontal cortices, the thalamus, the posterior limb of the internal capsule, the caudate nucleus and the coronal white matter. In conclusion, the measurements presented here are the first direct and quantitative demonstration of the presence of low-grade cerebral oedema in patients with hepatic encephalopathy. Further, absolute changes in tissue water content were quantified for various brain regions.


[Survey and budgetary impact of anaesthesia machines failures].
To assess the incidence and the causes of failures of anaesthesia machines in relation to aging. Study design. - Prospective survey from january 1996 to july 2000. The causes (mechanical or electronic), the moment of identification (checklist, maintenance operation or quality-control operation) of each anaesthetic machine failure, the repair cost and the maintenance cost of 14 anaesthetic machines have been collected and entered into a database. Over 31,948 anaesthesia delivered during the period of the study, 614 failures have been declared: 53% were related both to mechanical problems or monitoring failure and 40% were identified during the pre-operative checklist. In half of the cases, a specially trained anaesthetic nurse was able to correct the failure in the operating theatre. The annual rate of anaesthetic machine failure remained stable over the study period and the annual maintenance cost is approximatively 10% of the initial machine value. No procedure was cancelled because of a machine technical failure. Anaesthetic machine failure rate change according to the time should not be criteria for remplacement if rigorous quality control and maintenance operation are used.


The management of late radiation-induced rectal injury after treatment of carcinoma of the uterus.
Sixty-one of 1,418 (4.3 per cent) patients treated with radiation for carcinoma of the uterus from 1963 to 1983 had significant radiation-induced complications of the intestine develop which required a surgical opinion considering further management. Ninety-three per cent of these complications involved the rectum. Florid proctitis resolved within two years of onset in 33 per cent of the patients who were managed conservatively while 22 per cent of the patients died of disseminated disease within the same time period. Surgical treatment was eventually necessary in 39 per cent of the patients who were initially treated conservatively for radiation induced proctitis. Rectal excision with coloanal sleeve anastomosis produced a satisfactory result in eight of 11 patients with severe radiation injury involving the rectum. The incidence of radiation-induced and malignant rectovaginal fistula were similar (1 per cent), but disease-induced symptoms tended to occur earlier after primary treatment (a median of eight months) compared with radiation-induced symptoms (a median of 16 months).


Specific reactivity of fluorescein isothiocyanate-conjugated separated IgG and IgA from celiac disease sera on human tissues.
Study of the reaction of celiac disease-related antibodies with human substrates has been hampered by the immunological cross-reactivity between fluorescein isothiocyanate-conjugated anti-human immunoglobulins and immunoglobulins normally present in human tissues. In order to overcome this problem we extracted IgG from 2 celiac disease sera (positive for specific IgG and IgA antibodies) using a genetically engineered recombinant form of streptococcal protein G covalently immobilized on 4% agarose beads. The separated IgG and IgA was conjugated with fluorescein isothiocyanate and used in direct immunofluorescence on 0 blood group human duodenum, liver, and myocardium. Staining of the lamina propria beneath the villous epithelium, the endomysium of smooth muscle layers in the duodenum, and the liver sinusoidal and pericellular myocardial matrix was observed. The reactivity of celiac disease-related antibodies with various human tissues indicates that these antibodies are the result of a systemic immune response directed against all tissues containing matrix proteins.


Effects of a hurricane on growth and morbidity in children from low-income families in Kingston, Jamaica.
This is the first report of the effects of a hurricane on children's health and nutritional status in which data were available preceding and following the event. When Hurricane Gilbert struck Jamaica in 1988, a longitudinal study was in progress in which children's weights and heights were recorded every 2 months and their morbidity histories taken every week. The investigation included 127 stunted (low height-for-age) and 32 non-stunted children aged 23-44 months, living in poor areas of Kingston. The data from the 4 months before and after the hurricane were compared. There was an increase in the occurrence of respiratory symptoms including rapid or difficult breathing (P < 0.04), coughs (P < 0.001) and nasal discharges (P < 0.001) during the first 2-month period after the hurricane. However there was no significant effect on the occurrence of diarrhoea and injuries. Deficits were also found in height gain (P < 0.001) during the same period. These adverse effects were found in spite of the large amount of food aid received and the aggressive health education programme implemented after the hurricane.


Long-term paracorporeal ventricular support systems: a single-center experience.
The Berlin Heart EXCOR is a first-generation paracorporeal, pneumatic ventricular assist device that creates pulsatile flow. It can be used for long-term support of the left and/or right ventricule during end-stage heart failure. The aim of this study was to share our clinical experience in 54 patients. Between April 2007 and August 2012, 54 patients with end-stage heart failure underwent Berlin Heart EXCOR ventricular assist device implantation, including 5 females and 9 children. Twenty-four patients (44%) were in Intermacs level 1, 11 (21%) in level 2, and 19 (35%) in level 3. Biventricular support was applied to 13 patients. Device implantation was performed with an "on pump" beating heart technique while 6 other patients underwent intervention operations while the aortic valve has under cross-clamp. Tricuspid annuloplasty was performed in 6 patients. There was no peroperative death. Nine patients (17%) underwent re-exploration because of hemorrhage in the early postoperative period. Heart transplantation was performed in 32 patients (59%), while 10 (19%) are still under pump support with a mean follow-up of 13 months. Although 1 was successfully weaned from the system, 11 patients (20%) died during the support. Pump-head exchange was required 19 times in 17 patients because of visible thrombus or fibrin deposit in the pump head or due to membrane rupture. The use of long-term paracorporeal assist devices has decreased in recent years because of the increased popularity of implantable devices that permit longer survival and a better quality of life. We believe that the Berlin Heart EXCOR has a special role because it can be used in pediatric patients and especially in critical conditions like Intermacs levels 1 and 2.


Dietary fluoride intake of 15-19-year-old male adults residing in the United States.
The average daily dietary fluoride intakes of 15-to-19-year-old males were estimated from the analysis of 24 FDA "market basket" food collections made from 1975 to 1982. The data indicate that 15-to-19-year-old males residing in fluoridated (greater than 0.7 ppm) cities had an average daily dietary fluoride intake of 1.85 mg/day when the diet provided an estimated caloric intake of 11.72 megajoules (2800 calories). In non-fluoridated cities, with less than 0.3 ppm in the drinking water, the average dietary fluoride intake was 0.86 mg/day. The beverages and drinking water contributed an average of 75 +/- 2% of the daily dietary fluoride intake.


Glycosidic constituents from Carpesium cernuum L.
Carpesides A (1) and B (2), two new 8-O-4'-neolignan glucosides, and eupatriol 9-O-beta-d-apiofuranosyl-(1 --> 6)-beta-d-glucopyranoside (3), one new monoterpenoid diglycoside, together with 10 known glycosidic compounds, were isolated from the aerial part of Carpesium cernuum L. The structures of the new glycosides are elucidated by means of chemical methods and spectroscopic studies.


Levels of trimethoprim and sulphamethoxazole in human seminal plasma.
An oral dose of the combination trimethophim/sulphamethoxazole (160 and 800 mg, respectively) gave concentrations of trimethoprim in seminal plasma that were equal to or higher than that in blood plasma 2--4 hours after intake. Trimethoprim was excreted by both the prostate and seminal vesicles. With a trimethoprim resistant strain it was possible to demonstrate in three patients with subclinical prostatitis that the concentrations of sulphamethoxazole two hours after the oral intake were 20, 26, and 74 microgram/ml in the blood plasma, respectively. This study was carried out on patients who had been on treatment for 2--5 days prior to investigation. In view of published reports on the pH of the fluids from the human prostate and seminal vesicles, respectively, and the importance of the hydrogen ion concentrations for the excretion of lipid soluble substances into the prostatic fluid it was unexpected to find similar concentrations of trimethoprim in both the "prostatic" and "vesicular" fluids. Preliminary studies on the pH of these fluids indicated, however, that they were in the same order of magnitude, i.e. 7.5 to 8.0. The multiglandular origin of the human seminal plasma must be considered in all studied concerning the excretion of drugs into the semen. Many problems related to the excretion of drugs, e.g. organ specificity, kinetics, and influence on organ and spermatozoal functions remain to be investigated.


The value of Wilms tumor susceptibility gene 1 in cytologic preparations as a marker for malignant mesothelioma.
It has been shown that detection of the Wilms tumor susceptibility gene 1 protein (WT1) has diagnostic utility in the distinction of mesothelioma from adenocarcinoma in tissue sections of pleural tumors. This immunohistochemical study evaluates the effectiveness of WT1 as a marker for malignant mesothelioma in paraffin sections of cell block preparations derived from effusion specimens. The authors evaluated 111 cell blocks for WT1 immunoreactivity, including 14 mesotheliomas and 97 metastatic adenocarcinomas from various sites. Nuclear reactivity for WT1 was observed in all samples of mesothelioma. However, only 22 of 97 samples (23%) of metastatic adenocarcinoma, nearly all of which were of ovarian origin (91%), exhibited nuclear reactivity for WT1. In 14 other samples (most of pulmonary derivation), WT1 staining restricted to the cytoplasm was observed for some tumor cells and was regarded as nonspecific. Based on this staining profile, WT1 represents an effective marker for mesotheliomas in cell block preparations and can aid in its distinction from pulmonary adenocarcinoma. In assessment of effusion specimens with metastatic carcinoma, nuclear reactivity for WT1 is highly suggestive of an ovary primary tumor


Comparison of indirect immunofluorescence (IF) test with enzyme-linked immunosorbent assay (ELISA) in screening of hybridomas to very virulent Marek's disease virus.
For identifying virus-specific antigens of Marek's disease virus (MDV), monoclonal antibodies (MAbs) against strain Md5 of serotype 1, which is known to be a very virulent MDV (vvMDV), were isolated. Fifty-eight hybridoma clones that secreted MAbs against vvMDV were obtained. Of these MAbs, 36 gave positive reactions in an immunofluorescence (IF) test, and 22 gave positive reactions on enzyme-linked immunosorbent assay (ELISA). None of these MAbs gave positive reactions in both the IF test and ELISA. Of the MAbs that gave positive reactions in the IF test, 33 clones reacted with MDV1-specific epitopes, the other three reacting with MDV1-HVT intertypic epitopes. None of the clones reacted with MDV1-MDV2 intertypic epitopes. Three virus-specific polypeptides were identified by radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) or immunoblotting. These polypeptides were recognized by 12 MAbs giving positive reactions by IF, but by none of those giving positive reactions by ELISA. In addition, size heterogeneity of the MDV1-specific phosphorylated polypeptides in the MDV1 strains was shown using the MAbs against Md5.


The impact of reconstruction algorithms on semi-automatic small lesion segmentation for PET: a phantom study.
A robust lesion segmentation method is critical for quantification of lesion activity in positron emission tomography (PET), especially for the cases where lesion boundary is not discernible in the corresponding computed tomography (CT). However, lesion delineation in PET is a challenging task, especially for small lesions, due to the low intrinsic resolution, image noise and partial volume effect. The combinations of different reconstruction methods and post-reconstruction smoothing on PET images also affect the segmentation result significantly which has always been overlooked. Therefore, the aim of this study was to investigate the impact of different reconstruction methods on semi-automated small lesion segmentation for PET images. Four conventional segmentation methods were evaluated including region growing technique based on maximum intensity (RGmax) and mean intensity (RGmean) thresholds, Fuzzy c-mean (FCM) and watershed (WS) technique. All these methods were evaluated on a physical phantom scan which was reconstructed with Ordered Subset Expectation Maximization (OSEM) with Gaussian post-smoothing and Maximum a Posteriori (MAP) with quadratic prior respectively. The results demonstrate that: 1) the performance of all the segmentation methods subject to the smoothness constraint applied on the reconstructed images; 2) FCM method applied on MAP reconstructed images yielded overall superior performance than other evaluated combinations.


Experimental partitioning of rainfall into throughfall, stemflow and interception loss by Haloxylon ammodendron, a dominant sand-stabilizing shrub in northwestern China.
Rainfall partitioning by the vegetation canopy represents a significant component of the local hydrological cycle by reshaping the amount and spatial distribution of rainfall. Measuring the components of rainfall partitioning, however, has been a challenging task due to laborious- and time-consuming field experiments. In this study, to probe the influences of long-term afforestation on dynamic patterns of rainfall partitioning, the dominant sand-stabilizing shrub Haloxylon ammodendron at three different ages was selected for field measurements during the 2020-2021 growing season. The throughfall percentage for young H. ammodendron (YH, 75.9 %) was significantly higher than that for middle-aged H. ammodendron (MAH, 63.4 %) and mature H. ammodendron (MH, 62.4 %) (p < 0.05 for all cases). However, the interception loss percentage of YH (22.3 %) was significantly lower than that for MAH (35.0 %) and MH (36.5 %) (p < 0.05 for all cases). No significant difference was found for stemflow percentage among YH (1.8 %), MAH (1.5 %) and MH (1.1 %). Smaller rainfall events contributed to a higher interception loss percentage and a lower net rainfall percentage for all ages. Both throughfall and stemflow percentage first showed increasing trends and then tended to be stable with increasing rainfall amount and duration, whereas interception loss percentage showed the opposite patterns. Rainfall partitioning was significantly correlated with the plant area index, stem basal area and canopy height (p < 0.05 for all cases), which may account for significant differences in rainfall partitioning patterns, as all shrubs experienced the same weather conditions. The average funneling ratio was 56.6, 26.7 and 17.9 for YH, MAH and MH, respectively. These results suggested that H. ammodendron afforestation can have a significant impact on rainfall partitioning by reducing net rainfall reaching the soil and may have some implications for local water budget and ecosystem management in oasis-desert ecotones.


Copying holograms using light of reduced spatial and temporal coherence.
A reduced spatial and temporal coherence light from a halogen lamp is used to copy holographic optical elements, and a maximum diffraction efficiency of 80% is achieved.


Bedside whole-blood clotting times: validity after snakebites.
Venomous snakebites contribute to morbidity and mortality throughout the world, most commonly in resource-poor areas, with about 2.5 million humans sustaining snakebites annually. Coagulopathy is a significant cause of both morbidity and mortality in these patients. In the absence of more sophisticated hematological studies or obvious physical findings, many clinicians must rely on whole-blood clotting times to assess whether their patients are coagulopathic. Alternative (bedside) methods to assess clotting times are often officially recommended and used, but have not been validated. We assessed two bedside methods for measuring whole-blood clotting time after snakebites for their congruence with results from a hospital laboratory. Over a 5-month period, 46 sequential patients presenting with a possible snakebite had blood drawn for bedside (using syringe and ceftriaxone bottle as containers) and laboratory whole-blood clotting tests. All three tests used ∼5 mL whole blood and looked for any clot formation within 20 min. Compared to the laboratory, the syringe method correctly classified the patients 84.7% of the time (sensitivity 88.9%; specificity 82.4%). The bottle method correctly classified the patients 86.8% of the time (sensitivity 83.3%; specificity 90.0%). Comparing the area under the Receiver Operator Characteristics curves shows that both the syringe and bottle methods do not differ in their discrimination for identifying clotting. Both the syringe and ceftriaxone bottle bedside clotting test methods appear to be accurate enough to help guide therapy after potential snake envenomations when formal laboratory testing is unavailable.


Galectin signature in normal pregnancy and preeclampsia.
Members of the galectin family are expressed within the female reproductive tract and have been shown to be involved in multiple biological functions that support the progression of pregnancy. Specific expression patterns of different members of this family have been identified at the maternal decidua and on the placental side. In some cases, mechanisms by which galectins exert their functions have been delineated in adverse pregnancy outcomes. This review summarizes studies on galectins that have been documented to be important for pregnancy maintenance, either supporting the maternal adaptation to pregnancy or the placentation process. In addition, we focus our discussion on the role of galectins in preeclampsia, a specific life-threatening pregnancy disorder.


Bilateral abducens paralysis revealing bipartite spheno-occipital synchondrosis fracture in a child: case based update.
After reviewing the literature about sella turcica and spheno-occipital synchondrosis fracture, we present the case of a 6-year-old girl who suffered complex sphenoid bone fracture produced by an indirect mechanism. The girl fell from her height after a blunt head injury due to a falling lamppost. Computer tomography (CT) scan showed occipital, sellar and clival fractures with pneumatocephalus. The probable fracture mechanism is contre-coup injury linked to osseous and brain oscillations after head trauma. Two complications, abducens nerve palsy and cerebrospinal fluid fistula, are observed and discussed.


Synthesis, crystal structure and Hirshfeld analysis of triphenylphosphine-(4-bromophenyl)borane.
The title compound, alternatively called (4-bromophenyl)dihydrido(triphenylphosphanyl)boron, C24H21BBrP, crystallizes in the space group P-1 with one symmetry-independent molecule in the asymmetric unit. The B and P atoms both adopt the expected tetrahedral geometry. A possible intramolecular B-Hδ-...δ+H-C dihydrogen bond exists between an arene group on phosphorus and the borane center. In the solid state, the title molecule is connected to its neighbors by intermolecular C-H...π interactions, with the arene rings on the P atom acting as hydrogen-bond donors and the arene ring of the borane acting as the acceptor. Hirshfeld analysis supports the existence of these interactions, as well as weak Br...Br interactions between neighboring molecules.


Rhizobial acyl carrier proteins and their roles in the formation of bacterial cell-surface components that are required for the development of nitrogen-fixing root nodules on legume hosts.
Acyl carrier protein (ACP) of Escherichia coli is a small acidic protein which functions as carrier of growing acyl chains during their biosynthesis and as donor of acyl chains during transfer to target molecules. This unique ACP of E. coli is expressed constitutively. In more complex bacteria, multiple ACPs are present, indicating a channeling of pools of multi-carbon units into different biosynthetic routes. In rhizobia, for example, besides the constitutive ACP (AcpP) involved in the biosynthesis and transfer of common fatty acids, three specialized ACPs have been reported: (1) the flavonoid-inducible nodulation protein NodF, (2) AcpXL that transfers 27-hydroxyoctacosanoic acid to a sugar backbone during lipid A biosynthesis, and (3) the RkpF protein which is required for the biosynthesis of rhizobial capsular polysaccharides. All three of those specialized rhizobial ACPs are required for the biosynthesis of cell-surface molecules that play a role in establishing the symbiotic relationship between rhizobia and their legume hosts. Surprisingly, the recently sequenced genomes from Mesorhizobium loti and Sinorhizobium meliloti suggest even more candidates for ACPs in rhizobia.


Processing redundant signals: coactivation, divided attention, or what?
The evidence for and against a redundancy gain in reaction time (RT) when the target is repeated in the visual display is reviewed. We consider the relevance of redundancy gains under these circumstances to the question of whether attention can be simultaneously directed to separate locations in the visual field. In the present experiments, two capital letters were the target stimuli in a two-alternative forced-choice RT paradigm. In addition to the usual conditions of single-target trials, trials on which the target is repeated in the display, and trials on which the target occurs with a noise letter, we introduced the innovation of a condition in which both targets occur in the display. In our two experiments, RT was fastest with single-target displays and slowest with displays containing a target and a noise letter. There was no significant difference in RT to displays in which the target was repeated and displays in which both targets were presented. Both conditions showed a redundancy gain when compared with displays containing a target and a noise letter. The lack of response competition in the both-targets condition and the overall pattern of the results were well explained by a unitary attentional focus that serially processed the letters in the display. Analyses of minima and maxima RTs were consistent with this interpretation.


The hepatoprotective effect of putrescine against cadmium-induced acute liver injury.
The hepatoprotective effect of putrescine against cadmium liver injury was investigated. Male Wistar rats were injected with a dose of cadmium (6.5 mg CdCl(2)/kg bodyweight, intraperitoneally). Normal saline (group I) or putrescine (300 micro mol/kg bodyweight; group II) were injected 2, 5 and 8 h later. A number of animals of both groups were killed 0, 12, 16, 24, 48 or 60 h after cadmium intoxication. Liver tissue was histologically assessed for necrosis, apoptosis, peliosis, mitoses, and inflammatory infiltration. Apoptosis was also quantified by the TUNEL assay for hepatocytes and nonparenchymal liver cells. The discrimination between hepatic cell subpopulations was achieved histochemically. The mitotic index in hematoxylin-eosin-stained sections and by the immunochemical detection of Ki67 nuclear antigen, (3)H-thymidine incorporation into hepatic DNA, and hepatic thymidine kinase activity were all used as indices of liver regeneration. Both hepatocyte apoptosis and liver necrosis evolved in a biphasic temporal pattern. Nonparenchymal cell apoptosis and peliosis hepatis evolved in a monophasic pattern and were correlated closely. Putrescine administration totally reversed liver necrosis and hepatocyte apoptosis. The time profile of nonparenchymal apoptosis was altered and peliosis hepatis was also totally attenuated. In conclusion, putrescine protected hepatocytes and modulated the mechanism of cadmium-induced acute hepatotoxicity.


Stomach microbiota, Helicobacter pylori, and group 2 innate lymphoid cells.
The stomach has been thought to host few commensal bacteria because of the existence of barriers, such as gastric acid. However, recent culture-independent, sequencing-based microbial analysis has shown that the stomach also harbors a wide diversity of microbiota. Although the stomach immune system, especially innate lymphoid cells (ILCs), has not been well elucidated, recent studies have shown that group 2 ILCs (ILC2s) are the dominant subtype in the stomach of both humans and mice. Stomach ILC2s are unique in that their existence is dependent on stomach microbiota, in sharp contrast to the lack of an impact of commensal microbiota on ILC2s in other tissues. The microbiota dependency of stomach ILC2s is partly explained by their responsiveness to interleukin (IL)-7. Stomach ILC2s express significantly higher IL-7 receptor protein levels on their surface and proliferate more in response to IL-7 stimulation in vitro than small intestinal ILC2s. Consistently, the stomach expresses much higher IL-7 protein levels than the small intestine. IL-5 secreted from stomach ILC2s promotes immunoglobulin (Ig) A production by plasma B cells. In a murine model, stomach ILC2s are important in containing Helicobacter pylori infection, especially in the early phase of infection, by promoting IgA production.


Use of a lentiviral flap vector for induction of CTL immunity against melanoma. Perspectives for immunotherapy.
A central triple-stranded DNA structure created during HIV-1 reverse transcription, the central flap, acts as a cis-active nuclear import determinant of the HIV-1 DNA genome. Insertion of the sequences responsible for formation of the central DNA flap into an HIV-1-derived vector strongly enhances its transduction efficiency. HIV-1 vectors with or without inclusion of the DNA flap and encoding the same melanoma polyepitope were constructed to transduce dendritic cells (DCs) and to evaluate their capacity for induction of melanoma-specific cytotoxic T-lymphocyte (CTL) responses ex vivo and in vivo. HIV-1 vectors including the DNA flap transduced up to 100% of immature mouse and human DCs. Inoculation of HLA-A2.1 transgenic mice with this flap vector elicited vigorous and multi-specific long-term anti-melanoma CTL responses, whereas the parental vector lacking the flap sequence was less efficient. Furthermore, human DCs transduced ex vivo with the recombinant DNA flap vector displayed efficient multi-specific primary human CTL responses against melanoma. Lentiviral vectors including the DNA flap should be powerful tools both for active immunization and for the ex vivo priming of CTL for tumor immunotherapy.


Increased level of advanced oxidation protein products in patients with coronary artery disease.
Witko-Sarsat et al. previously reported that advanced oxidation protein products (AOPP), which were determined by measurements of optical density (OD) at 340 nm under acidic conditions, were present at high levels in the plasma of uremic patients. We measured AOPP in the plasma of 392 patients who underwent coronary angiography because of suspected coronary artery disease (CAD) and examined the association between plasma AOPP levels and CAD. We also collected plasma from hemodialysis patients and healthy volunteers for AOPP assay. Plasma fractionation by size-exclusion chromatography was performed to determine the weights of the molecules involved in the AOPP activity. The elution patterns obtained after plasma fractionation by size-exclusion chromatography were the similar in both CAD and hemodialysis patients, but differed from those of the healthy volunteers, whose AOPP levels were low. We also found that, the severity score of CAD obtained by the Gensini scoring system had good correlation with AOPP quartiles. Multivariable regression models revealed that the plasma AOPP level was significantly related to CAD status. This is the first report of an association between AOPP and CAD. AOPP was an independent risk factor for CAD. Oxidative stress is speculated to be associated with the pathogenesis of CAD.


Publisher Correction: Captain GBP1: inflammasomes assemble, pyroptotic endgame.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Parallel MRI at microtesla fields.
Parallel imaging techniques have been widely used in high-field magnetic resonance imaging (MRI). Multiple receiver coils have been shown to improve image quality and allow accelerated image acquisition. Magnetic resonance imaging at ultra-low fields (ULF MRI) is a new imaging approach that uses SQUID (superconducting quantum interference device) sensors to measure the spatially encoded precession of pre-polarized nuclear spin populations at microtesla-range measurement fields. In this work, parallel imaging at microtesla fields is systematically studied for the first time. A seven-channel SQUID system, designed for both ULF MRI and magnetoencephalography (MEG), is used to acquire 3D images of a human hand, as well as 2D images of a large water phantom. The imaging is performed at 46 mu T measurement field with pre-polarization at 40 mT. It is shown how the use of seven channels increases imaging field of view and improves signal-to-noise ratio for the hand images. A simple procedure for approximate correction of concomitant gradient artifacts is described. Noise propagation is analyzed experimentally, and the main source of correlated noise is identified. Accelerated imaging based on one-dimensional undersampling and 1D SENSE (sensitivity encoding) image reconstruction is studied in the case of the 2D phantom. Actual threefold imaging acceleration in comparison to single-average fully encoded Fourier imaging is demonstrated. These results show that parallel imaging methods are efficient in ULF MRI, and that imaging performance of SQUID-based instruments improves substantially as the number of channels is increased.


[Dietary management of food allergies].
A therapeutic diet is only indicated if the diagnosis of food allergy is certain. In the case of an unproven suspicion everything should be done to confirm or disprove the diagnosis by diagnostic measures (patient history, diet history, food and symptom protocol etc.). In the case of proven food allergy there are different aims of nutritional therapy. Most importantly the patient should learn how to identify his eliciting agent in pre-packed and non-prepacked food and how to avoid it. The elimination diet should take into account the individual condition of the patient. In particular, if basic foods have to be eliminated from the diet, safe alternatives have to be determined to maintain a nutritionally balanced diet and to maintain a high quality of life. To ensure the nutritional adequacy of the diet, micro- and macronutrient intake should be analysed and improved, if necessary. For pollen-associated food allergy and certain chronic conditions it might be necessary to be aware of possible triggering factors as well as potential cross-reactions. Moreover, individual tolerance should be maintained during elimination diets and not jeopardized by voluntary and unnecessary self-denial of certain foods.An effective individual management of food allergies by the patient is considerably dependent on the quality of the nutritional education and therapy.


Acute Myocarditis with Infarct-like Presentation in a Pediatric Population: Role of Cardiovascular Magnetic Resonance.
Chest pain is a typical symptom of acute myocarditis in adolescents. It may be indistinguishable from myocardial ischemia so it is called "infarct-like pattern." Cardiovascular magnetic resonance has an important role as a non-invasive diagnostic tool. The aim of our study is to provide a description of an acute myocarditis series with infarct-like pattern and to evaluate the cardiovascular magnetic resonance role in a pediatric population. We included all pediatric patients (0-16 years) admitted to our hospital (May 2007-May 2016) with clinical diagnosis of acute myocarditis and infarct-like presentation (chest pain, EKG alterations, and released cardiac biomarkers). Diagnosis was confirmed with cardiovascular magnetic resonance using Lake Louise criteria. Seven patients (five males, two females) with a median age of 14 years (12.5-15.2) were included. All patients showed ST-segment changes and released cardiac biomarkers. Three patients had left ventricular hypertrophy and two presented mild systolic left ventricular dysfunction. All patients had at least two positive Lake Louise criteria. Late gadolinium enhancement was positive in all of them. With a median follow-up of 23 months (8-47), all of them are alive, with no cardiac symptoms and normal ventricular function. Infarct-like pattern is a typical presentation of acute myocarditis in adolescents. CMR should be performed in this population and may be considered as a first-line diagnostic tool. Its high sensitivity in infarct-like acute myocarditis may allow us to avoid endomyocardial biopsy. Unlike what was described in adults, late gadolinium enhancement does not imply worse outcome in our series.


Radionuclide release from simulated waste material after biogeochemical leaching of uraniferous mineral samples.
Biogeochemical mineral dissolution is a promising method for the released of metals in low-grade host mineralization that contain sulphidic minerals. The application of biogeochemical mineral dissolution to engineered leach heap piles in the Elliot Lake region may be considered as a promising passive technology for the economic recovery of low grade Uranium-bearing ores. In the current investigation, the decrease of radiological activity of uraniferous mineral material after biogeochemical mineral dissolution is quantified by gamma spectroscopy and compared to the results from digestion/ICP-MS analysis of the ore materials to determine if gamma spectroscopy is a simple, viable alternative quantification method for heavy nuclides. The potential release of Uranium (U) and Radium-226 ((226)Ra) to the aqueous environment from samples that have been treated to represent various stages of leaching and passive closure processes are assessed. Dissolution of U from the solid phase has occurred during biogeochemical mineral dissolution in the presence of Acidithiobacillus ferrooxidans, with gamma spectroscopy indicating an 84% decrease in Uranium-235 ((235)U) content, a value in accordance with the data obtained by dissolution chemistry. Gamma spectroscopy data indicate that only 30% of the (226)Ra was removed during the biogeochemical mineral dissolution. Chemical inhibition and passivation treatments of waste materials following the biogeochemical mineral dissolution offer greater protection against residual U and (226)Ra leaching. Pacified samples resist the release of (226)Ra contained in the mineral phase and may offer more protection to the aqueous environment for the long term, compared to untreated or inhibited residues, and should be taken into account for future decommissioning.


Are measures of body habitus associated with mortality in cystic fibrosis?
Nutrition is an important component of clinical care for patients with cystic fibrosis. We aimed to test the hypothesis that increased BMI, height, and level of creatinine as a biomarker for lean muscle mass are associated with lower mortality and whether differences in these measures may contribute toward sex differences in survival in cystic fibrosis. Using a cohort study design, we analyzed data from the UK Cystic Fibrosis Registry for patients who attended an annual assessment visit in 2007 and were followed-up until July 2009. Of 1,517 individuals, 62 died during the follow-up period. The odds of death were higher among patients in the lowest quintile of serum creatinine compared with the rest of the study population (OR, 3.28; 95% CI, 1.79-5.98). Increased height and higher BMI were also associated with lower risk of death. The higher mortality in female patients (OR, 1.48; 95% CI, 0.93-2.34) was reversed by adjustment using the absolute values for height, BMI, and serum creatinine level (adjusted OR, 0.44; 95% CI, 0.21-0.90) but not by the use of sex-specific values for these exposure variables. Lower muscle mass, shorter stature, and a low BMI are associated with increased mortality in cystic fibrosis. These measures of body habitus may contribute to the sex-specific survival differences in individuals with cystic fibrosis.


Production of thromboxanes by transformed lymphocytes--effect on heart contractility.
We studied the effect of transformed lymphocytes from patients with chronic lymphocytic leukaemia (CLL) and the Raji cell (Raji) on the response of rat isolated atria to sodium arachidonate (AA). In contrast to normal lymphocytes, CLL cells and Raji cells decrease the contractile tension of rat isolated atria. Addition of exogenous AA (1.98 X 10(-6) M) to Raji, further reduced the isometric developed tension. Time of culture of Raji was important, as the negative inotropic effect was greater at 72 h than at 24 h of culture. Living cells were required and cell-free supernatants were inactive. Preincubation of CLL cells or Raji with cyclooxygenase inhibitors (acetyl salycilic acid, indomethacin) or inhibitors of thromboxane (TX) synthesis (imidazole, L-8027) abolished the negative inotropic response suggesting the contribution of TXs. L-8027 also reduced the growth rate of Raji cells, indicating that TXs may play a role in the regulation of cell division. The production of TXs by CLL and Raji cells from both endogenous and exogenous sources provided additional support to this hypothesis and suggested that activation of this metabolic pathway may be related to cell transformation.


Low-risk Prostate Cancer: Identification, Management, and Outcomes.
The incidence of low-risk prostate cancer (PCa) has increased as a consequence of prostate-specific antigen testing. In this collaborative review article, we examine recent literature regarding low-risk PCa and the available prognostic and therapeutic options. We performed a literature review of the Medline, Embase, and Web of Science databases. The search strategy included the terms: prostate cancer, low risk, active surveillance, focal therapy, radical prostatectomy, watchful waiting, biomarker, magnetic resonance imaging, alone or in combination. Prospective randomized trials have failed to show an impact of radical treatments on cancer-specific survival in low-risk PCa patients. Several series have reported the risk of adverse pathologic outcomes at radical prostatectomy. However, it is not clear if these patients are at higher risk of death from PCa. Long-term follow-up indicates the feasibility of active surveillance in low-risk PCa patients, although approximately 30% of men starting active surveillance undergo treatment within 5 yr. Considering focal therapies, robust data investigating its impact on long-term survival outcomes are still required and therefore should be considered experimental. Magnetic resonance imaging and tissue biomarkers may help to predict clinically significant PCa in men initially diagnosed with low-risk disease. The incidence of low-risk PCa has increased in recent years. Only a small proportion of men with low-risk PCa progress to clinical symptoms, metastases, or death and prospective trials have not shown a benefit for immediate radical treatments. Tissue biomarkers, magnetic resonance imaging, and ongoing surveillance may help to identify those men with low-risk PCa who harbor more clinically significant disease. Low-risk prostate cancer is very common. Active surveillance has excellent long-term results, while randomized trials have failed to show a beneficial impact of immediate radical treatments on survival. Biomarkers and magnetic resonance imaging may help to identify which men may benefit from early treatment.


A new pentanuclear cyano-bridged complex [(Fe(II)(tetraazamacrocycle))3{mu-NC-FeIII(CN)5)2]: synthesis, spectroscopic and magnetic characterization.
Hexacyanoferrate(III) reacts with [FeII(meso)(CH3CN)2](ClO4)2.2CH3CN (meso=5,5,7,12,12,14-hexamethyl-1,4,8,11-tetraazacyclotetradecane) in acetonitrile/water mixture producing the title complex, where three [Fe(meso)]2+ units are connected by two [Fe(CN)6](3-) anions. Molecular modeling (MM+) shows a fairly linear molecule and Mössbauer data are consistent with two terminal pentacoordinated low spin iron(II)-meso units linked to one hexacoordinated low spin iron(II)-meso through two hexacoordinated low spin iron(III) units. Spectroscopic characterization showed a typical mixed-valence charge transfer band and the degree of electron coupling was calculated to be H(AB)=678 cm(-1). Magnetic properties exhibited an antiferromagnetic exchange interaction between the iron(III) ions with a coupling constant J= -44 cm(-1).


The anti-tumor effects of CT-26 derived exosomes enriched by MicroRNA-34a on murine model of colorectal cancer.
As conventional therapeutics failed to provide satisfied outcomes against one of the most prevalent cancers, colorectal cancer (CRC), we purposed to implicate MicroRNA (miR)-34a, as a major tumor suppressor, to be delivered by tumor-derived exosomes (TEXs) and investigated its anti-tumor functions in-vivo. TEXs were isolated from CT-26 cell line and loaded with miR-34a mimic. Then, mice bearing CRC were treated with miR-34a-enriched TEX (TEX-miR-34a) and then examined for the relative tumor-suppressive impacts of the TEX as well as its potential in promoting an anti-tumor immune response. TEX-miR-34a significantly reduced tumor size and prolonged survival of mice bearing CRC. TEX-miR-34a was able to diminish gene expressions related to invasion, angiogenesis and immune evasion. It was also capable of inducing T cell polarization toward CD8+ T subsets among tumor-infiltrating lymphocytes, draining lymph nodes (DLNs) and spleen cells. Moreover, cytotoxic T cells were professionally induced in mice receiving TEX-miR-34a and the secretion of interleukin (IL)-6, IL-17A and tumor necrosis factor (TGF)-β was reduced in DLNs. However, the enhanced levels of interferon-γ were evaluated in DLN and spleen displaying the polarization of anti-tumor immune responses. Interestingly, mice receiving TEX alone showed a noticeable reduction in certain oncogenic gene expressions as well as IL-17A secretion in DLNs. TEX-miR-34a demonstrated the potential to induce beneficial anti-tumor immune responses and TEXs, aside from the delivery function of miRNA, revealed certain anti-tumor beneficial characteristics which could introduce TEX-miR-34a as a promising approach in CRC combination therapies.


Genetics of Gastric Cancer.
Gastric cancer represents a major cause of cancer mortality worldwide despite a declining incidence. New molecular classification schemes developed from genomic and molecular analyses of gastric cancer have provided a framework for understanding this heterogenous disease, and early findings suggest these classifications will be relevant for designing and implementing new targeted therapies. The success of targeted therapy and immunotherapy in breast cancer and melanoma, respectively, has not been duplicated in gastric cancer, but trastuzumab and ramucirumab have demonstrated efficacy in select populations. New markers that predict therapeutic response are needed to improve patient selection for both targeted and immunotherapies.


A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates.
Matriptase is an epithelial-derived type-II transmembrane serine protease. This protease is expressed prominently in the villus tip of small-intestinal epithelia at which senescent cells undergo shedding and/or apoptosis. The basement membrane of epithelial cells, including small-intestinal epithelial cells, contains extracellular matrix (ECM) proteins such as fibronectin and laminin. We found previously that high concentrations of a recombinant matriptase catalytic domain (r-MatCD) (e.g. 1 μM) caused an increased detachment of and increases in the activity of apoptotic effector caspase-3 in a rat small-intestinal epithelial IEC-6 line cultured on laminin-coated plates and proposed that at sites with its high level of expression, matriptase contributes to promoting shedding and/or detachment-induced death of epithelial cells through a mechanism mediating loss of cell-ECM adhesion. In this study, we found that even without increasing cell detachment, a high concentration of r-MatCD causes an increase in caspase-3 activity in IEC-6 cells cultured on fibronectin-coated plates, suggesting that the recombinant matriptase can cause apoptosis by a mechanism unrelated to cell detachment. Also, r-MatCD-treated IEC-6 cells on fibronectin were found to display spindle-like morphological changes. We suggest that r-MatCD causes apoptosis of IEC-6 on fibronectin by a mechanism involving the disruption of cell integrity.


Hydrogen-Bonding Motifs in Adducts of Allylamine with the 10 Simplest n-Alcohols: Single-Crystal X-ray Diffraction Studies and Computational Analysis.
In this paper, we analyzed the homologous series of 10 allylamine adducts with n-alcohols from methanol to decanol. These are the first adduct structures containing aliphatic n-alcohols and an aliphatic amine as co-formers. While all of the ingredients are liquids under ambient conditions, the phases were synthesized with the use of the in situ crystallization technique assisted by IR laser-focused radiation at atmospheric pressure. The structures were characterized by single-crystal X-ray diffraction. All of the phases contain the amine and alcohol in a 1:1 ratio. The architecture of the structures, based on hydrogen-bonding interactions between NH2 and OH moieties, depends on the size of the alcohol and changes in a systematic way. The three smallest alcohol adducts contain centrosymmetric layers of molecules of the L4(4)8(8) type. The next four alcohol adducts have the T4(2) topology. The structures with the biggest alcohols contain non-centrosymmetric L6(6) layers. The structural investigations were supported by periodic DFT calculations at the B3LYP/pobTZVP level. The cohesive and adhesive energies made up of layer (E lbe) and ribbon (E rbe) binding energies were used to predict which type of architecture can be formed. The thermal stabilities of the adducts correlate with the melting points of the co-forming alcohols, with no evident relation to the adduct architecture.


Peptide-based affinity media for solid-phase extraction of Ochratoxin A from wine samples: Effect of the solid support on binding properties.
A suitable sample clean up is a key point in the development of an analytical method. Peptide-based affinity media have recently gained attention in the selective extraction of defined target analytes from complex samples. In this paper we investigated the thermodynamic and kinetic binding properties of different stationary phases (Amberlite IRC-50, Lewatit CNP105, Toyopearl CM-650 M, porous silica gel beads and micrometric glass beads) functionalized with a hexapeptide sequence binding the Ochratoxin A. The highest values of the equilibrium binding constant (Keq) and binding site concentration (Bmax) were obtained for Lewatit CNP105 (Keq: 98.1×10(6) M(-1), Bmax: 30.8 μmol/g), followed by Toyopearl and micrometric glass beads, whereas the worst performances were obtained with Amberlite IRC-50 and porous silica gel beads. Also kinetic performances show the same trend. These results highlight that the surface chemical nature has a key role in the binding properties of solid supports used as affinity media for the selective extraction of well-defined target molecules. Finally, Lewatit CNP105 was compared with Amberlite IRC-50 as solid support in SPE extraction of OTA from 14 wine samples fortified with OTA at 2 and 4 μg l(-1) levels. The extracts were analyzed by HPLC with fluorescence detection (λexc 333 nm, λem 460 nm) showing no significant matrix effects, with a LOD and LOQ of 0.45 and 1.5 μgl(-1), respectively, and good recoveries between 71% and 108% for Amberlite IRC-50 and 91% and 101% for Lewatit CNP105. While both supports showed a statistically comparable extraction accuracy, a statistically significant difference was found in terms of extraction precision, confirming that the solid phase based on Lewatit CNP105 performs better than the solid phase based on Amberlite IRC-50.


Identification of three novel LRRK2 mutations associated with Parkinson's disease in a Calabrian population.
LRRK2 mutations are common in familial and sporadic Parkinson's disease (PD) cases. We present a screening of the most frequently mutated exons of LRRK2 in Calabrian population. Eighty-eight PD patients diagnosed according to standard criteria, underwent screening for LRRK2 mutations in exons 19, 21, 24, 25, 27, 29, 31, 32, 33, 35, 38, 40, 41, and 48. Eight LRRK2 variations were identified in nine patients affected by PD, including three novel missense variations (p.Phe1227Leu, p.Gly1520Ala, p.Ile2020Ser) and five previously identified mutations (p.Ala1151Thr, IVS31+3A>G, p.Arg1514Gln, p.Gly2019Ser, p.Thr2356Ile). LRRK2 frequency mutations were approximately 10.2% in all PD patients, 12% in familial, 8% in sporadic cases. The p.Gly2019Ser mutation was found in 2.3% of the total cohort and in 3.2% of sporadic cases. The clinical features of LRRK2-associated with PD in our patients were similar to those of idiopathic PD although most LRRK2 mutated patients presented with bradykinesia instead of tremor; 33.3% developed dementia. We identified three novel LRRK2 mutations and reported a higher frequency in Calabria compared to previously reported data possibly due to the relative genetic isolation of the Calabrian population. These findings contribute to the understanding of the role of LRKK2 variations in PD and provide additional genetic insight into this disease.


Malleability of the folding mechanism of the outer membrane protein PagP: parallel pathways and the effect of membrane elasticity.
Understanding the interactions between membrane proteins and the lipid bilayer is key to increasing our ability to predict and tailor the folding mechanism, structure and stability of membrane proteins. Here, we have investigated the effects of changing the membrane composition and the relative concentrations of protein and lipid on the folding mechanism of the bacterial outer membrane protein PagP. The folding pathway, monitored by tryptophan fluorescence, was found to be characterized by a burst phase, representing PagP adsorption to the liposome surface, followed by a time course that reflects the folding and insertion of the protein into the membrane. In 1,2-dilauroyl-sn-glycero-3-phosphocholine (diC(12:0)PC) liposomes, the post-adsorption time course fits well to a single exponential at high lipid-to-protein ratios (LPRs), but at low LPRs, a second exponential phase with a slower folding rate constant is observed. Interrupted refolding assays demonstrated that the two exponential phases reflect the presence of parallel folding pathways. Partitioning between these pathways was found to be modulated by the elastic properties of the membrane. Folding into mixed 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine:diC(12:0)PC liposomes resulted in a decrease in PagP adsorption to the liposomes and a switch to the slower folding pathway. By contrast, inclusion of 1,2-dilauroyl-sn-glycero-3-phosphoserine into diC(12:0)PC liposomes resulted in a decrease in the folding rate of the fast pathway. The results highlight the effect of lipid composition in tailoring the folding mechanism of a membrane protein, revealing that membrane proteins have access to multiple, competing folding routes to a unique native structure.


Anticonvulsant properties of hypothermia in experimental status epilepticus.
Status epilepticus in patients often does not respond to first-line anticonvulsants, and subsequent treatment escalation with continuous intravenous anesthetics may be associated with significant side-effects. Therefore, alternative treatment regimens are urgently needed. Hypothermia has been shown to reduce excitatory transmission and may thus serve as an interesting adjunct in the management of status epilepticus. In the current experiments, three treatment groups were compared. Animals with self-sustaining status epilepticus were treated with external cooling for 3 h, with low-dose diazepam, or with a combination of both. The effect of these regimens on epileptic activity was compared with untreated controls. Animals that underwent cooling were rewarmed, and all animals were monitored for 5 h to assess occurrence and severity of motor seizures and frequency and amplitude of spontaneous epileptic discharges. Cooling alone significantly reduced number and severity of motor seizures but did not alter epileptic discharges. Cooling in addition to low-dose diazepam significantly diminished amplitudes and frequencies of epileptic discharges, while diazepam alone had only a minor reducing effect on discharge amplitudes. However, at later stages of status epilepticus, diazepam significantly reduced motor seizures. Following rewarming, the discharge frequency tended to increase again, suggesting partial reversibility. The current experiments show that in status epilepticus hypothermia exhibits anticonvulsant effects which are most pronounced if co-administered with low-dose diazepam. The results still require confirmation in other animal models and also clinical studies are urgently needed. However, our data indicate that cooling could well become a future adjunct in the treatment of status epilepticus in patients.


Pediatric trauma care: an overview of pediatric trauma systems and their practices in 18 US states.
The aim of this study was to describe the state of pediatric trauma system development in the United States in 1997 and 1998 and to characterize the hospitalization patterns of injured children in states with different types of pediatric trauma systems. The authors also investigated the impact of sociodemographic, injury, and geographic characteristics on those hospitalization patterns. The authors combined statewide hospital discharge data on hospitalized trauma patients less than 15 years old with data from the American Hospital Association, the Area Resource File, the Office of Management and Budget, the states' Departments of Health, and the US Census. Besides conducting descriptive analyses, the authors evaluated the role of several parameters in determining the likelihood of treatment in trauma and nontrauma centers using multivariate multinomial logistic regression models. There were 15 states with adult and pediatric trauma designation systems; 9 of them had statewide hospital discharge data available. In these 9 states, 77% of the discharges were from trauma centers with no pediatric designation. More severely injured children and children with injuries to the head, face, thorax, and abdomen were more likely to be discharged from trauma centers, although large percentages of these children were treated in nontrauma centers. Older children and children with commercial insurance were less likely to be treated at trauma centers, even when injury severity, body region injured, and other factors were accounted for. Even in states with trauma systems, a large proportion of severely injured children are treated in nontrauma center facilities.


Properties of a reconstituted calcium pump.
1. The translocation of 45Ca2+ in vesicles reconstituted with purified Ca2+ ATPase of sarcoplasmic reticulum and phospholipids was dependent on ATP and varied greatly with the composition of the phospholipids. 2. In contrast to sarcoplasmic reticulum fragments, the reconstituted vesicles were impermeable to 14C-labeled oxalate, 3H- or 32P-labeled ATP, or 32P-i. There was no translocation of phosphate from gamma-labeled ATP during Ca2+ uptake. These results are inconsistent with some current formulations of the mechanism of pump action. 3. Reversal of the Ca2+ pump and generation of ATP and ADP and P-i was observed when vesicles loaded with Ca2+ were exposed to ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid. 4. Experiments on the formation of phosphoenzyme with 32P-labeled ATP showed that most if not all functional ATPase molecules in the reconstituted vesicles were oriented in the same direction, as in the case of sarcoplasmic reticulum fragments.


Ketosis treatment and milk yield in dairy cows related to milk acetoacetate levels.
Milk yield and milk acetoacetate (M-acac) were measured weekly for the first 6 weeks of lactation in 5 herds with a ketosis problem. Ketosis treatments and the corresponding ketotest score, were also recorded. The treatment rate was highest 7-16 days after calving. Most of these early cases were associated with low ketone levels in milk, whereas the treatment rate for cows with high ketone levels was highest 17-31 days after calving. Nearly half of the treated cows were low-ketone animals. They were classified as ketosis cases in the cow health card records, although probably suffering from other post partum disturbances in many instances. About 40% of the cows with high ketone levels recovered spontaneously. Reduction in milk yield associated with peak M-acac levels was transient and moderate. It was concluded that health card statistics overestimates the severity of the ketosis problem in Norway.


The potential role of juvenile hormone acid methyltransferase in methyl farnesoate (MF) biosynthesis in the swimming crab, Portunus trituberculatus.
Juvenile hormone (JH) and methyl farnesoate (MF) play essential roles in the development and reproduction of insects and crustaceans respectively. Juvenile hormone acid methyltransferase (JHAMT) catalyzes the methyl esterification in insect JH biosynthesis, while the corresponding step in crustacean MF biosynthesis was long thought to be catalyzed by farnesoic acid O-methyltransferase (FAMeT). However, the new discovery of JHAMT orthologs in crustaceans indicates that JHAMT may also play essential role in the MF biosynthesis in crustaceans. Here we cloned and characterized the full-length cDNA encoding JHAMT in the swimming crab Portunus trituberculatus (PtJHAMT). Sequence and structure analysis of PtJHAMT revealed that it was composed of a 6-stranded β sheet with 9 α helices, and contained a signature Sadenosyl-L-methionine (SAM) binding motif, which is the hallmark in all SAM dependent methyltransferases (SAM-MTs). Several active sites that are critical for the interaction of SAM and JH/FA substrate were also conserved in PtJHAMT. The gene expression of PtJHAMT was highly specific to the mandibular organ, which is the sole site of MF synthesis. PtJHAMT expression significantly increased in the late-vitellogenic stage and mature stage, which suggests a possible role of PtJHAMT in modulating ovarian development. The role of PtJHAMT and PtFAMeT in MF biosynthesis was further investigated by RNA interfering (RNAi). Injection of PtJHAMT and PtFAMeT dsRNA both led to a decrease in hemolymph MF titers. Injection of PtHMGR dsRNA caused the decrease in PtJHAMT expression, but had no effect on mRNA level of PtFAMeT. Together these results suggested that JHAMT and FAMeT are both involved in the MF biosynthesis in crustaceans, while the JHAMT is highly specific to FA substrate, and FAMeT may have more catalytic functions.


PI3K p110β subunit in leptin receptor expressing cells is required for the acute hypophagia induced by endotoxemia.
Hypophagia and increased energy expenditure under inflammatory conditions, such as that observed after bacterial lipopolysaccharide (LPS) administration, are associated with leptin secretion. The hypophagic effect of leptin depends in part on the activation of PI3K signaling pathway. However, the role of PI3K in the endotoxemia-induced hypophagia has not been determined. In an attempt to examine the functional contribution of the PI3K pathway in hypophagia and weight loss induced by LPS (100 ug/Kg, ip), we performed a central pharmacological PI3K inhibition (LY294002). Additionally, to gain mechanistic insights on the role of the catalytic PI3K p110α subunit in leptin responsive cells, mice expressing Cre-recombinase driven by the Lepr promoter (LepR-Cre) were crossed with mice carrying a loxP-modified p110α allele (Pi3kca gene) (LepR(Δp110α)). As studies have suggested that the PI3K p110β subunit has a dominant role over p110α in energy homeostasis, we further crossed LepR-Cre mice with loxP-modified p110α and p110β (Pi3kcb gene) alleles (LepR(Δp110α+β)). In order to verify the requirement of leptin in PI3K effects on food intake, we also used leptin-deficient ob/ob mice. We found that LPS stimulates PI3K and STAT3 signaling pathways in cells expressing the leptin receptor. Central PI3K inhibition prevented LPS-induced hypophagia and weight loss. Genetic deletion of p110α subunit selectively in LepR cells had no effect on LPS-induced hypophagia and weight loss. However, p110α and p110β double deletion in LepR cells prevented LPS-induced hypophagia and partially reversed the weight loss. Leptin deficiency blunted LPS-induced acute pAKT and pSTAT3 phosphorylation and the acute suppression of food intake. Our studies show that the PI3K p110β subunit in LepR cells is required for acute endotoxemic hypophagia. The data provide promising approaches for PI3K inhibition in preventing low energy balance and cachectic states during inflammatory challenges.


Solution NMR backbone assignment of the SASH1 SLy proteins associated disordered region (SPIDER).
SASH1 is a scaffold protein with context-dependent biological functions in cell adhesion, tumor metastasis, lung development, and pigmentation. As a member of the SLy protein family, it contains the conserved SLY, SH3, and SAM domains. The 19 kDa SLY domain harbors over 70% of the SASH1 variants associated with pigmentation disorders. However, its solution structure or dynamics have not been investigated yet, and its exact position in the sequence is not clearly defined. Based on the bioinformatic and experimental evidence, we propose renaming this region to the SLy Proteins Associated Disordered Region (SPIDER) and defining the exact position to be amino acids 400-554 of SASH1. We have previously identified a variant in this region linked to a pigmentation disorder, S519N. Here, we used a novel deuteration technique, a suite of TROSY-based 3D NMR experiments, and a high-quality HNN to obtain near complete solution backbone assignment of SASH1's SPIDER. A comparison with the chemical shifts of non-variant (S519) SPIDER shows that the S519N substitution does not alter the free form solution structural propensities of SPIDER. This assignment is the first step to characterize the role of SPIDER in SASH1-mediated cellular functions and provides a model for the future study of sister SPIDER domains in the SLy protein family.


GPR110 (ADGRF1) mediates anti-inflammatory effects of N-docosahexaenoylethanolamine.
Neuroinflammation is a widely accepted underlying condition for various pathological processes in the brain. In a recent study, synaptamide, an endogenous metabolite derived from docosahexaenoic acid (DHA, 22:6n-3), was identified as a specific ligand to orphan adhesion G-protein-coupled receptor 110 (GPR110, ADGRF1). Synaptamide has been shown to suppress lipopolysaccharide (LPS)-induced neuroinflammation in mice, but involvement of GPR110 in this process has not been established. In this study, we investigated the possible immune regulatory role of GPR110 in mediating the anti-neuroinflammatory effects of synaptamide under a systemic inflammatory condition. For in vitro studies, we assessed the role of GPR110 in synaptamide effects on LPS-induced inflammatory responses in adult primary mouse microglia, immortalized murine microglial cells (BV2), primary neutrophil, and peritoneal macrophage by using quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA) as well as neutrophil migration and ROS production assays. To evaluate in vivo effects, wild-type (WT) and GPR110 knock-out (KO) mice were injected with LPS intraperitoneally (i.p.) or TNF intravenously (i.v.) followed by synaptamide (i.p.), and expression of proinflammatory mediators was measured by qPCR, ELISA, and western blot analysis. Activated microglia in the brain and NF-kB activation in cells were examined microscopically after immunostaining for Iba-1 and RelA, respectively. Intraperitoneal (i.p.) administration of LPS increased TNF and IL-1β in the blood and induced pro-inflammatory cytokine expression in the brain. Subsequent i.p. injection of the GPR110 ligand synaptamide significantly reduced LPS-induced inflammatory responses in wild-type (WT) but not in GPR110 knock-out (KO) mice. In cultured microglia, synaptamide increased cAMP and inhibited LPS-induced proinflammatory cytokine expression by inhibiting the translocation of NF-κB subunit RelA into the nucleus. These effects were abolished by blocking synaptamide binding to GPR110 using an N-terminal targeting antibody. GPR110 expression was found to be high in neutrophils and macrophages where synaptamide also caused a GPR110-dependent increase in cAMP and inhibition of LPS-induced pro-inflammatory mediator expression. Intravenous injection of TNF, a pro-inflammatory cytokine that increases in the circulation after LPS treatment, elicited inflammatory responses in the brain which were dampened by the subsequent injection (i.p.) of synaptamide in a GPR110-dependent manner. Our study demonstrates the immune-regulatory function of GPR110 in both brain and periphery, collectively contributing to the anti-neuroinflammatory effects of synaptamide under a systemic inflammatory condition. We suggest GPR110 activation as a novel therapeutic strategy to ameliorate inflammation in the brain as well as periphery.


Effect of alcohol on emotional responses to stress.
When participating in tasks involving various probabilities of receiving an electric shock, men who had previously drunk alcohol maintained high levels of anxiety while anxiety decreased in men who had previously drunk a placebo.


Data-based psychiatric consultation: applying mainframe computer capability to consultation.
Critical, intertwined objectives for consultation psychiatry include 1) the development of data-based clinical practices, and 2) heightening the effectiveness of consultation. Toward these ends, the Consultation Psychiatry Service at the University of Minnesota has previously conducted systematic studies of consultation and established an ongoing data collection system for consultations. Both steps have relied extensively upon mainframe computer capability. This primary application of computer capability to the field of psychiatric consultation is reviewed by the authors.


Oscillatory correlates of auditory working memory examined with human electrocorticography.
This work examines how sounds are held in auditory working memory (AWM) in humans by examining oscillatory local field potentials (LFPs) in candidate brain regions. Previous fMRI studies by our group demonstrated blood oxygenation level-dependent (BOLD) response increases during maintenance in auditory cortex, inferior frontal cortex and the hippocampus using a paradigm with a delay period greater than 10s. The relationship between such BOLD changes and ensemble activity in different frequency bands is complex, and the long delay period raised the possibility that long-term memory mechanisms were engaged. Here we assessed LFPs in different frequency bands in six subjects with recordings from all candidate brain regions using a paradigm with a short delay period of 3 s. Sustained delay activity was demonstrated in all areas, with different patterns in the different areas. Enhancement in low frequency (delta) power and suppression across higher frequencies (beta/gamma) were demonstrated in primary auditory cortex in medial Heschl's gyrus (HG) whilst non-primary cortex showed patterns of enhancement and suppression that altered at different levels of the auditory hierarchy from lateral HG to superior- and middle-temporal gyrus. Inferior frontal cortex showed increasing suppression with increasing frequency. The hippocampus and parahippocampal gyrus showed low frequency increases and high frequency decreases in oscillatory activity. This work demonstrates sustained activity patterns during AWM maintenance, with prominent low-frequency increases in medial temporal lobe regions.


A new serially transplantable human prostatic cancer (HONDA) in nude mice.
A new serially transplantable human prostatic cancer (HONDA) in nude mice was established from a patient with metastatic prostate carcinoma. The tumor grows well in male nude mice. Doubling time of the tumor weight at passage #13 was 9.5 +/- 0.87 days (mean +/- SD). The tumor retains the original histological features of adenocarcinoma even after 6 years of continuous passage. High levels of human prostatic acid phosphatase were detected by radioimmunoassay in sera from the tumor-bearing mice. The tumor cells contain human prostate specific antigen. Electron microscopy showed particles resembling type A retroviruses in cisterns of endoplasmic reticulum, and particles resembling type C retroviruses in the intercellular space of the tumor cells. The tumor grew well in female mice treated with testosterone, but not in untreated female mice or castrated male mice.