Early diagnosis and successful treatment of acute cytomegalovirus encephalitis in a renal transplant recipient.
We report the case of a 40-year-old male HIV-negative renal transplant patient with allograft rejection and immunosuppressive therapy who presented with acute cytomegalovirus (CMV) encephalitis. CT and MRI of the brain were normal but EEG showed diffuse slowing and dysrhythmia. In cerebrospinal fluid (CSF) initially 81 cells/microliters were found and immunocytochemistry showed a decreased CD4/CD8 ratio and increased values of activated lymphocytes, natural killer cells and immunoglobulin-containing cells. CMV-specific IgM antibodies in CSF and serum, immunostaining of CMV antigen in CSF cells and virus culture from CSF and urine were negative. During the first 3 weeks of illness no intrathecal production of immunoglobulins could be detected. Early diagnosis of CMV encephalitis was made by in situ hybridization (ISH) on CSF cell preparations and the polymerase chain reaction (PCR) which was positive in CSF and blood. On day 26 diagnosis was confirmed by detection of CMV-specific intrathecal IgG production. The patient was treated with ganciclovir, anti-CMV immunoglobulins and intrathecal beta interferon. He recovered completely after 2 months. Our data demonstrate the usefulness of ISH and PCR in the early diagnosis of CMV encephalitis and perhaps may encourage the use of intrathecal beta interferon in other patients with this disease.


BAP1 mutant uveal melanoma is stratified by metabolic phenotypes with distinct vulnerability to metabolic inhibitors.
Cancer cell metabolism is a targetable vulnerability; however, a precise understanding of metabolic heterogeneity is required. Inactivating mutations in BRCA1-associated protein 1 (BAP1) are associated with metastasis in uveal melanoma (UM), the deadliest adult eye cancer. BAP1 functions in UM remain unclear. UM patient sample analysis divided BAP1 mutant UM tumors into two subgroups based on oxidative phosphorylation (OXPHOS) gene expression suggesting metabolic heterogeneity. Consistent with patient data, transcriptomic analysis of BAP1 mutant UM cell lines also showed OXPHOShigh or OXPHOSlow subgroups. Integrated RNA sequencing, metabolomics, and molecular analyses showed that OXPHOShigh BAP1 mutant UM cells utilize glycolytic and nucleotide biosynthesis pathways, whereas OXPHOSlow BAP1 mutant UM cells employ fatty acid oxidation. Furthermore, the two subgroups responded to different classes of metabolic suppressors. Our findings indicate that targeting cancer metabolism is a promising therapeutic option for BAP1 mutant UM; however, tailored approaches may be required due to metabolic heterogeneities.


Minimum requirements in infection control.
Infection control (IC) activities are facing new challenges, but the resources dedicated to IC are too frequently insufficient. Heterogeneity of resources among centres and countries is huge, a fact that at least partly explains the differences in the results obtained. In this article, we review and discuss the available recommendations for minimum requirements in IC related to organizational aspects, IC staffing and the training of these staff, ward staffing, structural issues, and microbiological support. A professional-based consensus on the minimum requirements for IC in European centres based on present challenges and societal demands is needed.


Chymase mediates mast cell-induced angiogenesis in hamster sponge granulomas.
We investigated the contribution of mast cell chymase in mast cell-dependent angiogenesis using the hamster sponge-implant model, where angiogenesis in the granulation tissue surrounding the subcutaneously implanted sponge was evaluated by measuring the hemoglobin content. Daily local injection of compound 48/80 (3-100 microg/site/day), a potent mast cell activator, induced formation of granulomas and angiogenesis in time- and dose-dependent manners. This angiogenic response was inhibited by chymase inhibitors including chymostatin (> or = 1 nmol/site/day), soybean trypsin inhibitor (SBTI; > or = 1.4 nmol/site/day) and lima bean trypsin inhibitor (LBTI; > or = 3.3 nmol/site/day), but not by a tryptase inhibitor like leupeptin (> or = 700 nmol/site/day). Although pyrilamine (> or = 2,580 nmol/site/day), a histamine H1 receptor antagonist, and protamine (300 microg/site/day) also inhibited angiogenesis, these effects were much less pronounced than those by chymase inhibitors. Furthermore, antigen-induced angiogenesis in hamsters pre-sensitized with ovalbumin was also inhibited by the chymase inhibitors by 60-70%. Our results suggest that chymase is a major mediator in mast cell-mediated angiogenesis.


Role of enteric plexuses in the competency of telescoped intestinal valves.
The mechanism of preventing intestinal regurgitation by means of a telescoped intestinal valve has been investigated in this study. Ten telescoped intestinal valves were uniformly created in dogs. A tolerance test to barium reflux was performed before the animals were sacrificed 16 weeks postoperatively, which yielded various barium tolerance values. Statistical analysis revealed that the differences in barium tolerance values in the uniformly constructed valves were closely related to the proportion of the enteric plexuses which contained ganglion cells with normal shapes (r = 0.97, p less than 0.001) and their intestinal segment coverage (r = 0.94, p less than 0.001), but no correlation was found between barium tolerance and the thickness of the valvular musculature (r = 0.04) or the length of the artificial valve (r = 0.39). The results in this study suggest that the intact enteric plexuses play an important role in modulating the competency of the telescoped intestinal valve.


Viruses, virulence and pathogenicity.
Pathogenicity is a complex process with stringent requirements of both the host cell and the infecting virion. Among these requirements are a port of entry into host cells, a means of replication for the virus, and a means by which infection damages host cells. Damage to the host can result from multiple mechanisms including transformation, suppression of cellular metabolism, apoptosis, autoimmune responses directed against infected or uninfected tissues, or by molecular mimicry. In the attempt to identify new associations between viral infection and disease, investigators should be mindful that variable host factors as well as viral infection may be required for pathogenesis. Efforts to associate specific viral infections with specific diseases may be obscured by final common pathways through which multiple agents damage host cells in similar ways.


The study of distance changes between lumbar bi-cortical pedicle screws and anterior large vessels in patients with lumbar spondylolisthesis.
This paper was a anatomical radiographic study of distance between lumbar bi-cortical pedicle screws (BPSs) and anterior large vessels (ALVs) in patients with lumbar spondylolisthesis, and to provide clinical basis for evaluating the safety of bi-cortical pedicle screw implantation during lumbar spondylolisthesis. Complete Computed tomography (CT) data of 104 patients with grade I lumbar spondylolisthesis (L4 52 and L5 52) and 107 non-spondylolisthesis patients (control group) were collected in this study. The distances between lumbar 4,5(L4,5) and sacrum 1(S1) BPSs and ALVs (abdominal aorta, inferior vena cava, left and right common iliac artery, internal and external iliac artery) were respectively measured at different transverse screw angles (TSAs) (L4:5°,10°; L5:10°,15°; S1:0°,5°,10°) and analyzed by SPSS (v25.0). There were three types of distances from the anterior vertebral cortex (AVC) to the ALVs (DAVC-ALV): DAVC-ALV N, DAVC-ALV ≥ 0.50 cm, and DAVC-ALV < 0.50 cm; these different distances represented non-contact, distant and close ALV respectively. We calculated the incidences of screw tip contacting large vessels at different TSAs and provided the appropriate angle of screw implantation. In non-spondylolisthesis group, in L4, the appropriate left TSA was 5°, and the incidence of the close ALV was 4.62%. In S1, the appropriate left TSA was 0° and the incidence of the close ALV was 22.4%, while the appropriate right TSA was 10° and the incidence of the close ALV was 17.8%. In L4 spondylolisthesis group, in L4, the appropriate left TSA was 5°, and the incidence of the close ALV was 3.8%. In L5 spondylolisthesis group, in S1, the appropriate left TSA was 0° and the incidence of the close ALV was 19.2%, while the appropriate right TSA was 10° and the incidence of the close ALV was 21.2%. The use of BPS was not appropriate on the right side of L4 or on the either side of L5 both in spondylolisthesis and control group. In patients with lumbar 4 spondylolisthesis, the incidences of screw tip contacting large vessels were less than the control group in both L4 and 5. In patients with lumbar 5 spondylolisthesis, the incidences of screw tip contacting large vessels were less than the control group in L5, while there were no significant difference in S1. It is very important that considering the anatomical relationship between the AVC and the ALVs while planning BPSs. The use of BPS does not apply to every lumbar vertebra. In patients with lumbar spondylolisthesis and non-spondylolisthesis patients, the incidences of screw tip contacting large vessels are different.


HLA studies in brucellar spondylitis.
Among the clinical manifestations of brucellosis, arthritis may occur in over one-third of the patients. Different articular syndromes have been well recognized: some are definitely infectious in nature, whereas others appear to be reactive. The possibility that B27 and B7-CREG antigens could predispose to the occurrence of brucellar spondylitis was investigated in 14 mestizo Peruvian patients. No association was found. Since there is a low frequency of B27 and B7-CREG in the control population, the possibility that a "native" antigen could be the predisposing one remains to be elucidated.


Interoceptive deficits differentiate suicide groups and associate with self-injurious thoughts and behaviors in a military sample.
Previous research shows that interoceptive deficits are associated with harmful behaviors such as nonsuicidal self-injury (NSSI), eating disorder pathology, and suicide attempts. The present study replicates and extends this area of research by examining the association between interoceptive deficits and suicidality in a military sample. In Study 1, respondents to an online survey (N = 134) answered self-report questionnaires related to interoceptive deficits. Study 2 consisted of a secondary data analysis of 3,764 military service members who had previously completed questionnaires on interoceptive indicators, NSSI, suicide thoughts and attempts, and other psychopathology. Study 1 demonstrated that our interoceptive deficits latent variable had adequate psychometric properties. In Study 2, multigroup confirmatory factor analysis showed that scores on the interoceptive deficits latent variable were highest among suicide attempters, lowest among those with no suicide history, and intermediary among participants who had thought about but not attempted suicide. The interoceptive deficits latent variable was more strongly related to NSSI and suicidality than were posttraumatic stress disorder symptoms, hopelessness, gender, and age. These results confirm-and extend to a military sample-previous research showing that interoceptive deficits can provide important information about suicide risk.


Patient factors predict complications after partial nephrectomy: validation and calibration of the Preoperative Risk Evaluation for Partial Nephrectomy (PREP) score.
To develop and validate the Preoperative Risk Evaluation for Partial Nephrectomy (PREP) score to predict the probability of major postoperative complications after partial nephrectomy (PN) based on patient comorbidities. The Premier Healthcare Database was used to identify patients who had undergone elective PN. Through review of International Classification of Diseases ninth revision codes, we identified patient comorbidities and major surgical complications (Clavien-Dindo Grade III-V). Multivariable logistic regression was used to identify predictors of major complications. We used half of the set as the training cohort to develop our risk score and the other half as a validation cohort. From 2003 to 2015, 25 451 PNs were performed. The overall rate of major complications was 4.9%. The final risk score consisted of 10 predictors: age, sex, congestive heart failure, coronary artery disease, chronic obstructive pulmonary disease, chronic kidney disease, diabetes, hypertension, obesity, and smoking. In the training cohort, the area under the receiver operating characteristic curve (AUC) was 0.75 (95% confidence interval [CI] 0.73-0.78), while the AUC for the validation cohort was 0.73 (95% CI 0.70-0.75). The predicted probabilities of major complication in the low- (≤10 points), intermediate- (11-20 points), high- (21-30 points), and very high-risk (>30 points) categories were 3% (95% CI 2.6-3.2), 8% (95% CI 7.2-9.2), 24% (95% CI 20.5-27.8), and 41% (95% CI 34.5-47.8), respectively. We developed and validated the PREP score to predict the risk of complications after PN based on patient characteristics. Calculation of the PREP score can help providers select treatment options for patients with a cT1a renal mass and enhance the informed consent process for patients planning to undergo PN.


Fabrication of functional fibronectin patterns by nanosecond excimer laser direct write for tissue engineering applications.
Laser direct write techniques represent a prospective alternative for engineering a new generation of hybrid biomaterials via the creation of patterns consisting of biological proteins onto practically any type of substrate. In this paper we report on the characterization of fibronectin features obtained onto titanium substrates by UV nanosecond laser transfer. Fourier-transform infrared spectroscopy measurements evidenced no modification in the secondary structure of the post-transferred protein. The molecular weight of the transferred protein was identical to the initial fibronectin, no fragment bands being found in the transferred protein's Western blot migration profile. The presence of the cell-binding domain sequence and the mannose groups within the transferred molecules was revealed by anti-fibronectin monoclonal antibody immunolabelling and FITC-Concanavalin-A staining, respectively. The in vitro tests performed with MC3T3-E1 osteoblast-like cells and Swiss-3T3 fibroblasts showed that the cells' morphology and spreading were strongly influenced by the presence of the fibronectin spots.


Two new species of Wockia heinemann (Lepidoptera: Urodidae) from Jamaica.
Wockia diabolica n. sp. and Wockia tetroidon n. sp. are described from female specimens collected from Jamaica. Both new species are distinguished from Wockia chewbacca Adamski, a Mexican species, in having a pair of signa within the corpus bursa of the female genitalia. Photographic images are provided for the imagoes, and illustrations of the genitalia are included. Distributions of these new taxa represent the first records of Wockia from West Indies.


A search for the "multiple sclerosis virus"--lack of effect of brain extracts on myelin development in chickens, mice and rats.
Attempts were made to transmit possible infectious agents from tissue of MS patients into three animal species, under conditions designed to enhance the development of such an agent. Myelination was monitored by measuring CNPase activity and myelin basic protein. Although no significant effects were observed, the usefulness of a new assay for CNPase was demonstrated. Nude mice were found to have a lower level of CNPase than their heterozygous littermates.


The ecology of testate amoebae (Protists) in sphagnum in North-western Poland in relation to peatland ecology.
We studied the relationship between testate amoebae (Protozoa) communities and the depth to the water table (DWT), pH, conductivity, and microhabitat type in Sphagnum dominated peatlands of north-western Poland and built predictive (transfer function) models for inferring DWT and pH based on the testate amoebae community structure. Such models can be used for peatland monitoring and paleoecology. A total of 52 testate amoebae taxa were recorded. In a redundancy analysis, DWT and pH explained 20.1% of the variation in the species data and allowed us to identify three groups of taxa: species that are associated with (1) high DWT and low pH, (2) low DWT and low pH, and (3) high pH and mid-range DWT. Our transfer function models allow DWT and pH to be estimated with mean errors of 9.89 cm and 0.71 pH units. The prediction error of the DWT model and the tolerance of the species both increase with increasing dryness. This pattern mirrors the ecology of Sphagnum mosses: Species growing in wet habitats are more sensitive to change in water table depth than the species growing in drier microhabitats. Our results are consistent with studies of testate amoeba ecology in other regions, and they provide additional support for the use of these organisms in paleoecological and biomonitoring contexts.


Contribution of circulating hemocytes to the regeneration of heavy ion beams (12C5+) irradiated hematopoietic organs in the silkworm, Bombyx mori, through the way of phagocytosis of injured cells after invasion.
Heavy ion beam irradiation has promising effects on tumor therapy. Our previous study using the domesticated silkworm, Bombyx mori, showed that this irradiation could seriously damage larval hematopoietic organs but they would regenerate later. In the in vitro irradiation, most hemocytes died when hematopoietic organs and wing discs connected with epidermis were directionally irradiated from epidermis to hematopoietic organ and then cultured so as to exclude circulating hemocytes. A few hemocytes had escaped irradiation according to extremely low hematopoiesis in vitro. Almost no hemocytes could incorporate BrdU at 60 h after irradiation, with which living and proliferating hemocytes are also labeled. In the absence of circulating hemocytes, the irradiation-escaped hemocytes in the organs were not enough for cleaning all dead cells because lots of small dead bodies remained in situ post-irradiation. After irradiating hematopoietic organs in larvae (in vivo irradiation), only a few apoptotic cells were found when given the same length of recovery time, and most hemocytes maintained normal morphology. Many hemocytes incorporated BrdU when tested at the same time as the in vitro irradiation but this number was lower than that measured for control organs. Circulating hemocytes, labeled by fluorescent microbeads through phagocytosis before irradiation, were found to have invaded the in vivo irradiated hematopoietic organs where they help the irradiation-escaped hemocytes to clear dead cells in the process of regeneration. Hematopoiesis of the regenerated hematopoietic organs did not fully recover to the level of the control organs according to the number of hemocytes produced in tissue culture. Some of the released hemocytes obviously underwent apoptosis, suggesting a far-reaching bystander effect of carbon ion beams irradiation on hemocytes inside. From these results, it is suggested that, together with irradiation-escaped hemocytes, the invaded circulating hemocytes took part in the regeneration of heavy ion beams irradiated hematopoietic organs through the way of phagocytosis of injured hemocytes in vivo.


Milk 99Tcm-EHIDA test for enterogastric bile reflux.
The study and clinical assessment of enterogastric bile reflux has been restricted for want of a simple non-invasive test for its detection and quantification. This paper describes such a test in which biliary excretion scintigraphy has been combined and a mild meal provocation. Two of 10 healthy volunteers studied showed probable reflux of approximately 5 per cent of total initial abdominal field activity. Among 73 patients studied, 37 patients showed definite reflux of up to 47 per cent. Reflux occurred in 19 of 22 post-gastric surgery patients and in 7 of 22 patients with peptic ulcer disease, gastritis or gastro-oesophageal reflux. None of 7 patients with 'non-specific' abdominal pain showed any reflux, but 11 of 22 patients with gallstone disease or previous cholecystectomy showed reflux of up to 35 per cent, including 9 of 11 patients with loss of gallbladder reservoir function.


Gene expression of ascorbic acid biosynthesis related enzymes of the Smirnoff-Wheeler pathway in acerola (Malpighia glabra).
The Smirnoff-Wheeler (SW) pathway has been proven to be the only significant source of l-ascorbic acid (AsA; vitamin C) in the seedlings of the model plant Arabidopsis thaliana. It is yet uncertain whether the same pathway holds for all other plants and their various organs as AsA may also be synthesized through alternative pathways. In this study, we have cloned some of the genes involved in the SW-pathway from acerola (Malpighia glabra), a plant containing enormous amount of AsA, and examined the expression patterns of these genes in the plant. The AsA contents of acerola leaves were about 8-fold more than that of Arabidopsis with 5-700-fold higher mRNA abundance in AsA-biosynthesizing genes. The unripe fruits have the highest AsA content but the accumulation was substantially repressed as the fruit transitions to maturation. The mRNAs encoding these genes showed correlation in their expression with the AsA contents of the fruits. Although very little AsA was recorded in the seeds the mRNAs encoding all the genes, with the exception of the mitochondrially located L-galactono-1,4-lactone dehydrogenase, were clearly detected in the seeds of the unripe fruits. In young leaves of acerola, the expression of most genes were repressed by the dark and induced by light. However, the expression of GDP-D-mannose pyrophosphorylase similar to that encoded by A. thaliana VTC1 was induced in the dark. The expressions of all the genes surged after 24h following wounding stress on the young leaves. These findings will advance the investigation into the molecular factors regulating the biosynthesis of abundant AsA in acerola.


Radiological diagnosis of inorganic lead poisoning.
A 43-year-old woman had abdominal pain and anemia due to inorganic lead poisoning. Plain abdominal x-ray films showed metallic densities within the lumen of the large bowel. The patient admitted that she used to chew the sheets of metallic lead enclosing wine bottle necks regularly. We report the case because of both the unusual radiographic findings and the exceptional source of lead intoxication.


Modulation of production of hepatitis B surface antigen by a human hepatoma cell line.
Three drugs were assayed for their capacity to inhibit hepatitis B surface antigen (HBsAg) production by the PLC/PRF/5 human hepatoma cell line. The effect on cell growth and HBsAg production of Cordycepin, 6-azauridine, and Hygromicin B is reported. Hygromicin B, a translation inhibitor unable to penetrate normal cells, greatly reduced HBsAg production by growing and confluent cells.


Microbial cell factories for bio-based biodegradable plastics production.
The misuse of petroleum-based plastics has resulted in serious environmental pollution and resource wastage. Biodegradable plastics can be used as green substitutes for traditional plastics. Here, we discuss the feasibility and technical bottlenecks in developing microbial cell factories for the production of biodegradable plastics from lignocellulosic wastes. First, we introduce the basic properties of the main biodegradable plastics on the market, including poly(lactic acid), poly(hydroxyalkanoate), and poly(butylene adipate-co-terephthalate). We then demonstrate the feasibility of synthesizing petroleum-based biodegradable plastic monomers from bio-based raw materials and propose strategies to further advance their commercial production through metabolic engineering and synthetic biology. We also analyze the main challenges facing the current development of bio-based biodegradable plastic biosynthesis technology. Finally, we discuss the current major lignocellulose bioconversion processes and explore way to further improve the utilization efficiency of the main carbohydrates in lignocellulosic hydrolysates by microorganisms, from the perspectives of sugar transport, sugar assimilation, and carbon catabolite inhibition.


Photoredox ketone catalysis for the direct C-H imidation and acyloxylation of arenes.
The photoexcited aryl ketone-catalyzed C-H imidation of arenes and heteroarenes is reported. Using 3,6-dimethoxy-9H-thioxanthen-9-one as a catalyst in combination with a bench-stable imidating reagent, C-N bond formation proceeds with high efficiency and a broad substrate scope. A key part of this method is that the thioxanthone catalyst acts as an excited-state reductant, thus establishing an oxidative quenching cycle for radical aromatic substitution. The synthetic potential of this photoexcited ketone catalysis is further demonstrated by application to the direct C-H acyloxylation of arenes.


Membrane-actin interactions in morphogenesis: Lessons learned from Drosophila cellularization.
During morphogenesis, changes in the shapes of individual cells are harnessed to mold an entire tissue. These changes in cell shapes require the coupled remodeling of the plasma membrane and underlying actin cytoskeleton. In this review, we highlight cellularization of the Drosophila embryo as a model system to uncover principles of how membrane and actin dynamics are co-regulated in space and time to drive morphogenesis.


Influence of Quiet Time on the Auditory Environment of Infants in the NICU.
To determine the influence of quiet time on the auditory environment of infants in the NICU and to compare the effect of quiet time by room type, bed type, and infant clinical acuity. Cross-sectional, descriptive comparison study. Sixty-six infants born at less than 38 weeks gestation who were between 32 and 40 weeks postmenstrual age. The auditory environments of infants in two level 3 NICUs within a pediatric hospital system in the Southeastern United States. Each NICU implemented quiet time for 4 hours per day. We assessed the auditory environment of the participants using Language Environmental Analysis technology. We used paired t tests to assess differences in the auditory environment during quiet versus nonquiet time and to compare the effect of quiet time by room type, bed type, and clinical acuity. During quiet time, the auditory environment of participants had 13% more silence, 17% fewer electronic sounds, 25% less speech, and 30% fewer words than during nonquiet time. We observed greater differences in quiet time versus nonquiet time for infants in open bays and incubators and infants who had greater acuity. Our results support the implementation of quiet time to increase silence and reduce exposure to electronic sounds for infants in the NICU. Additional research is necessary to further examine the effect of quiet time on the auditory environment of infants in the NICU with consideration of environmental and clinical variables.


A unitary mechanism underlies adaptation to both local and global environmental statistics in time perception.
Our duration estimation flexibly adapts to the statistical properties of the temporal context. Humans and non-human species exhibit a perceptual bias towards the mean of durations previously observed as well as serial dependence, a perceptual bias towards the duration of recently processed events. Here we asked whether those two phenomena arise from a unitary mechanism or reflect the operation of two distinct systems that adapt separately to the global and local statistics of the environment. We employed a set of duration reproduction tasks in which the target duration was sampled from distributions with different variances and means. The central tendency and serial dependence biases were jointly modulated by the range and the variance of the prior, and these effects were well-captured by a unitary mechanism model in which temporal expectancies are updated after each trial based on perceptual observations. Alternative models that assume separate mechanisms for global and local contextual effects failed to capture the empirical results.


Enlarged perivascular spaces and plasma Aβ42/Aβ40 ratio in older adults without dementia.
Dilation of perivascular spaces (PVS) in the brain may indicate poor fluid drainage due to the accumulation of perivascular cell debris, waste, and proteins, including amyloid-beta (Aβ). No prior study has assessed whether plasma Aβ levels are related to PVS in older adults without dementia. Independently living older adults (N = 56, mean age = 68.2 years; Standard deviation (SD) = 6.5; 30.4% male) free of dementia or clinical stroke were recruited from the community and underwent brain MRI and venipuncture. PVS were qualitatively scored and dichotomized to low PVS burden (scores 0-1,) or high PVS burden (score>1). Plasma was assayed using a Quanterix Simoa Kit to quantify Aβ42 and Aβ40 levels. A significant difference was observed in plasma Aβ42/Aβ40 ratio between low and high PVS burden, controlling for age (F[1, 53] = 5.59, p = 0.022, η2 = 0.10), with lower Aβ42/Aβ40 ratio in the high PVS burden group. Dilation of PVS is associated with a lower plasma Aβ42/Aβ40 ratio, which may indicate higher cortical amyloid deposition. Future longitudinal studies examining PVS changes, and the pathogenesis of AD are warranted.


Network motifs are enriched with transcription factors whose transcripts have short half-lives.
Network motifs, the recurring regulatory structural patterns in networks, can self-organize to produce networks because of the large ratio of genes to transcription factors (TFs) in genomes. We find a common design principle of these motifs: the TFs whose transcripts have short half-lives are significantly enriched in motifs and hubs. This enrichment becomes one of the driving forces for the emergence of the network scale-free topology, enables the network to adapt quickly to environmental changes and mitigates gene expression fluctuations. Motifs are classified into subtypes that are preferentially used in different cellular conditions.


Antipsychotic medications and drug-induced movement disorders other than parkinsonism: a population-based cohort study in older adults.
To study the relationship between initiating therapy with an antipsychotic medication and a subsequent new diagnosis of a drug-induced movement disorder other than parkinsonism in older adults with dementia. Retrospective, population-based cohort study. Ontario, Canada. Ontario residents aged 66 and older with a diagnosis of dementia newly started on treatment with typical or atypical antipsychotic therapy. Estimated relative risk of developing a drug-induced movement other than parkinsonism in the 1-year follow-up period after starting therapy with an antipsychotic medication. From April 1, 1997, to March 31, 2001, 21, 835 older adults with dementia who were newly started on antipsychotic medications were identified. Nine thousand seven hundred ninety subjects were started on atypical antipsychotics and 12,045 subjects started on typical antipsychotics. Demographic characteristics were similar between the groups. There were 5.24 cases of tardive dyskinesia (TD) or other drug-induced movement disorder per 100 person-years on therapy with a typical antipsychotic and 5.19 cases per 100 person-years on therapy with an atypical antipsychotic. The risk of developing drug-induced movement disorder while being treated with an atypical agent was not statistically different from that with a typical antipsychotic (relative risk=0.99, 95% confidence interval=0.86-1.15; P<.93). Older adults with dementia who are treated with typical or atypical antipsychotic therapy are at risk for developing TD and other drug-induced movement disorders.


Figure/ground segregation from temporal delay is best at high spatial frequencies.
Two experiments investigated the role of spatial frequency in performance of a figure/ground segregation task based on temporal cues. Figure orientation was much easier to judge when figure and ground portions of the target were defined exclusively by random texture composed entirely of high spatial frequencies. When target components were defined by low spatial frequencies only, the task was nearly impossible except with long temporal delay between figure and ground. These results are inconsistent with the hypothesis that M-cell activity is primarily responsible for figure/ground segregation from temporal delay. Instead, these results point to a distinction between temporal integration and temporal differentiation. Additionally, the present results can be related to recent work on the binding of spatial features over time.


[Hexachlorbenzene (HCB) induced porphyria in rats. Influence of HCB-metabolites on the biosynthesis of heme (author's transl)].
Female adult Wistar rats were fed with a diet containing 0.05% hexachlorobenzene (HCB) or its metabolites, pentachlorobenzene (PCB) and pentachlorphenole (PCP). These chlorinated aromatic hydrocarbons produced an increase in the liver cytochrome P-450 content in about the same degree, however, only the application of HCB showed an extremely high rise in the P-450 enzymatic activity expressed in terms of the O-dealkylation of 7-Ethoxycoumarine. No alteration was observed in the urinary porphyrin excretion in the PCB and PCP treated animals, whereas 60 days after the beginning of the HCB application a high level of porphyrins could be detected in the urine of the animals. It seems unlikely therefore that the HCB metabolites (PCB and PCP) are porphyrogenic agents. In addition, although induction of the liver cytochrome P-450 system was observed after PCP pretreatment of the rats over a period of 40 days, the consequent application of HCB did not influence the establishment of the experimental porphyria.


Metabolic engineering of riboflavin production in Ashbya gossypii through pathway optimization.
The industrial production of riboflavin mostly relies on the microbial fermentation of flavinogenic microorganisms and Ashbya gossypii is the main industrial producer of the vitamin. Accordingly, bioengineering strategies aimed at increasing riboflavin production in A. gossypii are highly valuable for industry. We analyze the contribution of all the RIB genes to the production of riboflavin in A. gossypii. Two important metabolic rate-limiting steps that limit the overproduction of riboflavin have been found: first, low mRNA levels of the RIB genes hindered the overproduction of riboflavin; second, the competition of the AMP branch for purinogenic precursors also represents a limitation for riboflavin overproduction. Thus, overexpression of the RIB genes resulted in a significant increase in riboflavin yield. Moreover, both the inactivation and the underexpression of the ADE12 gene, which controls the first step of the AMP branch, also proved to have a positive effect on riboflavin production. Accordingly, a strain that combines both the overexpression of the RIB genes and the underexpression of the ADE12 gene was engineered. This strain produced 523 mg/L of riboflavin (5.4-fold higher than the wild-type), which is the highest titer of riboflavin obtained by metabolic engineering in A. gossypii so far. Riboflavin production in A. gossypii is limited by a low transcription activity of the RIB genes. Flux limitation towards AMP provides committed substrate GTP for riboflavin overproduction without detrimental effects on biomass formation. A multiple-engineered Ashbya strain that produces up to 523 mg/L of riboflavin was generated.


Nitrogen dioxide inhalation induces genotoxicity in rats.
Nitrogen dioxide (NO(2)) is a ubiquitous reactive free-radical gas, which has been associated with momentary and chronic health effects. In the present study, comet, micronucleus (MN) and DNA-protein crosslinks (DPC) assays were used to investigate the genotoxicity following in vivo inhalation exposure of rats to NO(2). The results show that inhalation exposure of rats to NO(2) induced DNA strand breakage and the formation of DPC in the cells from various internal organs (brain, lung, liver, spleen, kidney and heart), as well as resulted in obvious increase of MN frequency in the bone marrow cells of rats. Furthermore, above genotoxic responses showed significant linear dose-dependent manners. These results implicate that NO(2) is a genotoxic agent and these observations are informative for understanding the mechanisms of adverse effects of nitrogen dioxide.


Environmental fate and chemistry of raloxifene hydrochloride.
Raloxifene hydrochloride is a selective estrogen receptor modulator (SERM) used for the prevention and treatment of osteoporosis in women. Excretion of raloxifene occurs through the feces of patients. Raloxifene has the potential to be discharged into waste treatment systems after therapeutic use. Raloxifene hydrochloride was investigated using a battery of studies designed to describe its physical/chemical characteristics and define its fate in the environment. The mean measured solubility of raloxifene hydrochloride (+/- standard deviation) was 345.2 +/- 15.6 microg/ml, 13.3 +/- 0.6 microg/ml, 0.9224 +/- 0.015 microg/ml, and 627.4 +/- 132.0 microg/ml in aqueous buffers at pH 5, 7, and 9 and in unbuffered water, respectively. Raloxifene exhibited a mean molar absorptivity of 34,000 and a wavelength absorbance maximum at 287 nm for pH 5 and 7 aqueous buffer solutions and 297 nm at pH 9. Mean measured Kow values were 516 +/- 17, 1,323 +/- 91, and 1,556 +/- 135 at pH 5, 7, and 9, respectively. After 5 d at 50 degrees C, raloxifene hydrolyzed 8.02, 10.61, and 23.81% in pH 5, 7, and 9 aqueous buffers, respectively. In a 28-d hydrolysis study at 25 degrees C, the calculated first-order hydrolysis rates were 6.92 x 10(-4), 1.70 x 10(-3), and 7.66 x 10(-3)/d, and the corresponding half-lives were 1,001, 410, and 90 d in pH 5, 7, and 9 aqueous buffers, respectively. Raloxifene sorbed significantly to sewage treatment solids with Freundlich isotherm adsorption coefficients K between 2,000 and 3,000. Raloxifene degraded rapidly in the presence of sewage solids. In a system containing 0.470 g/L sludge solids, the raloxifene biodegradation rate and half-life were 0.0966/h and 7.17 h, respectively. In a 28-d aerobic-aquatic biodegradation study containing 30 mg/L sludge solids, the raloxifene biodegradation rate and half-life were 0.0188/d and 37 d, respectively. Given the fate and behavior of raloxifene in these studies, it is anticipated that raloxifene would rapidly dissipate in the environment.


Distribution of polybrominated diphenyl ethers in Japanese autopsy tissue and body fluid samples.
Brominated flame retardants are components of many plastics and are used in products such as cars, textiles, televisions, and personal computers. Human exposure to polybrominated diphenyl ether (PBDE) flame retardants has increased exponentially during the last three decades. Our objective was to measure the body burden and distribution of PBDEs and to determine the concentrations of the predominant PBDE congeners in samples of liver, bile, adipose tissue, and blood obtained from Japanese autopsy cases. Tissues and body fluids obtained from 20 autopsy cases were analyzed. The levels of 25 PBDE congeners, ranging from tri- to hexa-BDEs, were assessed. The geometric means of the sum of the concentrations of PBDE congeners having detection frequencies >50 % (ΣPBDE) in the blood, liver, bile, and adipose tissue were 2.4, 2.6, 1.4, and 4.3 ng/g lipid, respectively. The most abundant congeners were BDE-47 and BDE-153, followed by BDE-100, BDE-99, and BDE-28+33. These concentrations of PBDE congeners were similar to other reports of human exposure in Japan but were notably lower than concentrations than those reported in the USA. Significant positive correlations were observed between the concentrations of predominant congeners and ΣPBDE among the samples analyzed. The ΣPBDE concentration was highest in the adipose tissue, but PBDEs were distributed widely among the tissues and body fluids analyzed. The PBDE levels observed in the present study are similar to those reported in previous studies in Japan and significantly lower than those reported in the USA.


Rise of pediatric surgery collaboratives to facilitate quality improvement.
Broad changes in pediatric surgical care delivery are limited by the rarity of pediatric surgical diseases and the geographic dispersion of pediatric surgical care across different hospital types. Pediatric surgical collaboratives and consortiums can provide the patient sample size, research resources, and infrastructure to advance clinical care for children with who require surgery. Additionally, collaboratives can bring together experts and exemplar institutions to overcome barriers to pediatric surgical research to advance quality surgical care. Despite challenges to collaboration, many successful pediatric surgical collaboratives emerged in the last decade and continue to push the field forward towards high-quality, evidence-based care and improved outcomes. This review will discuss the need for continued research and quality improvement collaboratives in pediatric surgery, identify challenges faced when building collaboratives, and introduce future directions to expand impact.


Direct inhibition of collagen production in vitro by diabetic rat serum.
Diabetes mellitus is associated with a generalized defect in connective tissue metabolism, including decreased growth, poor wound healing, and osteopenia. To determine the role of circulating factors in the etiology of these defects, we studied the effects of diabetic rat serum (DRS) on collagen, the major protein component of connective tissues. After preincubation of costal cartilage from hypophysectomized rats with experimental serum for 20 hours, [3H] proline was added for final four hours of incubation. Collagen and noncollagen protein were quantitated using purified bacterial collagenase. Compared to incubation of tissue in buffer without added serum, collagen production in cartilage incubated with 2% DRS was decreased by 23% (P less than .05), and with 4% serum by 88% (P less than .01). In contrast, serum from normal rats (NRS) increased collagen to 158% above buffer-incubated cartilage at 1.0% (P less than .02) and to 196% at 2% serum (P less than .01). Noncollagen protein production decreased below buffer only after addition of 2% or more DRS and increased above buffer after addition of 2% or more of NRS (178%, P less than .05). Addition of insulin at 10 and 100 mU/mL to DRS did not reverse defective collagen production, and addition of glucose (up to 900 mg/dL) or ketones (20 mmol/L) to NRS did not induce the changes in collagen production seen after addition of diabetic serum. Chromatographic separation of serum revealed that the inhibitory activity of DRS was in the high molecular weight fraction (less than 5000).(ABSTRACT TRUNCATED AT 250 WORDS)


Parent-Adolescent Conflict in African American Families.
Parent-adolescent conflict is frequent in families and has implications for youth adjustment and family relationships. Drawing on a family systems perspective, we examined mothers', fathers', and two adolescent-aged siblings' (50.5 % females) reports of parent-adolescent conflict in 187 African American families. Using latent profile analysis in the context of an ethnic homogeneous design, we identified three family types based on levels of and differences between parent and youth conflict reports: low conflict, father high conflict, and younger sibling high conflict. Compared to low conflict families, youth in younger sibling high conflict families reported more depressive symptoms and risky behaviors. The results for parents' acceptance revealed that, in comparison to low conflict families, older siblings in father high conflict families reported lower acceptance from mothers, and mothers in these families reported lower acceptance of their children; further, older siblings in younger sibling high conflict families reported less acceptance from fathers, and fathers in these families reported less acceptance of their children. Results underscore the significance of levels of and both differences between and direction of differences in parents' and youth's reports of their "shared" experiences, as well as the importance of examining the larger family contexts of dyadic parent-relationships.


Methyl 6-chloro-nicotinate.
The mol-ecule of the title compound, C(7)H(6)ClNO(2), is almost planar, with a dihedral angle of 3.34 (14)° between the COOMe group and the aromatic ring. In the crystal, the mol-ecules are arranged into (1[Formula: see text]2) layers by C-H⋯N hydrogen bonds and there are π-π stacking inter-actions between the aromatic rings in adjacent layers [centroid-centroid distance 3.8721 (4) Å].


Ultrastructure of serum high density lipoproteins: facts and models.
The complexity of the structure of plasma high density lipoproteins (HDL) has invited numerous approaches which have been directed at the study of the intact particles, their apolipoproteins and reassembled complexes. Parameters such as flotation and sedimentation coefficients, size and molecular weight have been determined and in addition, through scattering techniques, an understanding has been obtained on the long range organization between core (cholesteryl esters and triglycerides) and surface components (unesterified cholesterol, phospholipids and apoproteins). In the case of the apolipoproteins, the knowledge of their primary structure has facilitated the study of their physicochemical properties in solution and at the air-water interface and has also permitted realistic predictions of the two dimensional organization, not only of their alpha-helical segments but also of the beta-pleated sheets, random coil and beta-turns, all of which have amphipathic properties. When all of the information from the physical and chemical studies is put together, the various HDL can be described as spherical structures having a liquid core of radius, r - 20.2 A, surrounded by a monolayer of cholesterol and phospholipids with closely packed hydrophobic ends on the surface of the core. The organization of the apoproteins at the lipoprotein interface is comparatively less understood. However, reasonable predictions can be made on secondary structure considerations and on their behavior at the air-water interface. The emerging overall structural information can be translated into a space-filling model that not only provides a useful representation of HDL, but, more importantly, a basis for planning future studies on the elucidation of the structure of these particles on a molecular level.


Regeneration in the central nervous system of a pulmonate mollusc, Melampus.
The left cerebral ganglion was ablated from 72 anesthetized, adult Melampus bidentatus (Mollusca: Pulmonata). Skin incisions were well healed and normal feeding and locomotion observed four days after surgery. Dissections of animals sacrificed weekly showed that most nerves and connectives regrew within 30 days, attaching to the swollen end of the major labial nerve. The enlarged end of this nerve later developed into a distinctive bud; some of these buds contained cell bodies as soon as 42 days after surgery. As the first known report of central nervous tissue regeneration in molluscs, this study points to the need for controls in experiments involving section or ablation of nervous tissue in molluscs.


LDassoc: an online tool for interactively exploring genome-wide association study results and prioritizing variants for functional investigation.
Existing approaches to plot association results from genome-wide association studies (GWAS) are in the form of static Manhattan plots and often lack data integration with rich databases on variant regulatory potential as well as population-specific linkage disequilibrium patterns. We created an intuitive web module for uploading and efficiently exploring GWAS association results. Interactive plots and sortable tables allow researchers to query genomic regions of interest, facilitating the integration of data on linkage disequilibrium, variant regulatory potential and potential target genes. External links allow for visualization of association results in the UCSC genome browser as well as easy access to publically available databases (e.g. dbSNP and RegulomeDB). Through improved visualization and data integration, LDassoc offers genomic researchers a specialized environment to examine association signals and suggests variants for functional investigation. LDassoc is a free and publically available web tool which can be accessed online at https://analysistools.nci.nih.gov/LDlink/? tab=ldassoc. email@example.com.


Comparative study on pyrolysis of bamboo in microwave pyrolysis-reforming reaction by binary compound impregnation and chemical liquid deposition modified HZSM-5.
The deactivation of catalyst is a significant reason for its limited application during the catalytic fast pyrolysis (CFP) process. To reduce the coke formation, binary compound impregnation (BCI) and chemical liquid deposition (CLD) were used to modify HZSM-5 catalysts. At the same time, the self-designed microwave reactor separated the pyrolysis of bamboo and catalytic upgrading of primary vapor, which made the catalytic effect more thorough. Experimental results indicated that CLD used TiO2 deposition to cover external acid sites, while BCI by phosphorus-nickel could cover and partly destroy superficial acid sites through two different ways. Within the scope of the loaded amount studied, the yield of aromatic hydrocarbons in the oil phase increased at first and then decreased, while the coke formation reduced continuously. BTX (benzene, toluene and xylene), the most valuable product in bio-oil, drastically increased by 39.1% and 22.6% respectively over the CLD and BCI modified catalysts. Considering the catalytic performance as well as cost, CLD over HZSM-5 has more advantages in the CFP process to upgrade bio-oil.


Afferent connections of the median preoptic nucleus in the rat: anatomical evidence for a cardiovascular integrative mechanism in the anteroventral third ventricular (AV3V) region.
The afferent connections of cell groups in the anteroventral third ventricular (AV3V) region were investigated using wheat germ agglutinin-horseradish peroxidase conjugate injections into the AV3V area. The median preoptic nucleus was found to receive a unique set of inputs from the subfornical organ, the paraventricular nucleus of the hypothalamus, the parabrachial nucleus, the nucleus of the solitary tract and the ventrolateral medulla. As each of these structures has been implicated in various aspects of cardiovascular regulation, it is likely that the cardiovascular effects of AV3V lesions are due at least in part to median preoptic damage.


Mild electrical stimulation at 0.1-ms pulse width induces p53 protein phosphorylation and G2 arrest in human epithelial cells.
Exogenous low-intensity electrical stimulation has been used for treatment of various intractable diseases despite the dearth of information on the molecular underpinnings of its effects. Our work and that of others have demonstrated that applied electrical stimulation at physiological strength or mild electrical stimulation (MES) activates the PI3K-Akt pathway, but whether MES activates other molecules remains unknown. Considering that MES is a form of physiological stress, we hypothesized that it can activate the tumor suppressor p53, which is a key modulator of the cell cycle and apoptosis in response to cell stresses. The potential response of p53 to an applied electrical current of low intensity has not been investigated. Here, we show that p53 was transiently phosphorylated at Ser-15 in epithelial cells treated with an imperceptible voltage (1 V/cm) and a 0.1-ms pulse width. MES-induced p53 phosphorylation was inhibited by pretreatment with a p38 MAPK inhibitor and transfection of dominant-negative mutants of p38, MKK3b, and MKK6b, implying the involvement of the p38 MAPK signaling pathway. Furthermore, MES treatment enhanced p53 transcriptional function and increased the expression of p53 target genes p21, BAX, PUMA, NOXA, and IRF9. Importantly, MES treatment triggered G2 cell cycle arrest, but not cell apoptosis. MES treatment had no effect on the cell cycle in HCT116 p53(-/-) cells, suggesting a dependence on p53. These findings identify some molecular targets of electrical stimulation and incorporate the p38-p53 signaling pathway among the transduction pathways that MES affects.


Embryopathies due to spermatozoal impairment in Xenopus laevis.
An in vitro test system, involving gametes of Xenopus laevis has been used to study the effect of antifertility agents upon spermatozoa as manifest in developing embryos. Exposure to either the isomeric forms of dimethylmyleran, methyl methanesulphonate or gamma-radiation led to development of a range of embryopathies, whereas treatment with steroidal drugs or the rodent epididymal chemosterilants alpha-chlorohydrin and trimethylphosphate was compatible with production of apparently normal offspring.


miR-125b, a target of CDX2, regulates cell differentiation through repression of the core binding factor in hematopoietic malignancies.
MicroRNA-125b (miR-125b), a small noncoding RNA molecule, has been found to be deregulated and functions as an oncogene in many cancers including hematopoietic malignancies. However, the mechanisms accounting for miR-125b dysregulation remain to be elucidated. The present study aims to identify the factors that might contribute to up-regulation of miR-125b in human hematopoietic malignancies and its downstream targets for lineage-specific differentiation. We at first reported that CDX2, a homeobox transcription factor, binds to promoter regions of the miR-125b gene and activates transcriptional regulation of miR-125b in malignant myeloid cells. We further revealed that increasing levels of CDX2 in malignant myeloid cells activate miR-125b expression, which in turn inhibits core binding factor β (CBFβ) translation, thereby counteracting myeloid cell differentiation, at least for granulocytic lineage, and promoting leukemogenesis. Interestingly, we found that this novel pathway including CDX2, miR-125b, and CBFβ was mediated by undergoing all-trans-retinoic acid induction. Once differentiation ensues with all-trans-retinoic acid treatment, CDX2 activity decreases, leading to a reduction in miR-125b transcription and up-regulation of CBFβ in myeloid cells and in patients. The study provides a new mechanism that contributes to hematopoietic malignancies, which could involve deregulation of miR-125b and its up- and downstream factors. As altered expression of miRNAs has been reported in a wide range of malignancies, delineating the underlying molecular mechanisms of aberrant miRNA expression and characterizing the upstream and downstream factors will help to understand important steps in the pathogenesis of these afflictions.


Epidural and pericranial abscesses complicating frontal sinusitis in a comatose child.
Most complications of sinusitis have a significant risk of severe morbidity and possibly mortality. Therefore a keen awareness of the subtle signs and symptoms of bacterial spread beyond the paranasal sinuses must be maintained. This increased awareness is especially important in mentally retarded and psychomotor handicapped children because they can develop a rapidly fatal course despite the absence of any warning signs.


Internalization of human 5-HT4a and 5-HT4b receptors is splice variant dependent.
The family of 5-HT4 receptors comprises 16 putative splice variants. We have previously shown that there are differences in signal transduction of the h5-HT(4a) and h5-HT(4b) receptors. In the present study, the internalization of these two splice variants following receptor stimulation was investigated with confocal microscopy on living cells. Chimeric receptors, h5-HT(4a)-GFP and h5-HT(4b)-GFP were generated by fusing the coding sequence of the 5-HT4 receptor with the coding sequence of the GFP. The agonist stimulation of fluorescent receptors resulted in a time-dependent internalization of the h5-HT(4b)-GFP receptor, but not of the h5-HT(4a)-GFP receptor. The h5-HT(4b) receptor displays a dual coupling to G(alpha)i,o and G(alpha)s proteins, in contrast to the h5-HT4a receptor, which couples to Galphas proteins only. We investigated whether the difference in internalization of the two splice variant receptors was related to their differential coupling. Therefore, we performed agonist-stimulation of the receptor following inhibition of the G(alpha)i,o protein coupling using PTX. The h5-HT(4b) receptor internalization is PTX insensitive. We co-transfected the fluorescent chimeric receptors with other wild-type variants, which did not produce an alteration of the receptor trafficking. These findings provide the first evidence of differential internalization between the two splice variants, 5-HT(4a) and 5-HT(4b) receptors.


Proceedings from The 8th Annual International Society for Musculoskeletal Imaging in Rheumatology (ISEMIR) Conference.
The International Society for Musculoskeletal Imaging in Rheumatology (ISEMIR) was founded in 2005 with the goal of discussing matters related to imaging in rheumatology, particularly, validation, education, and use in both clinical practice and research. The field of musculoskeletal (MSK) imaging is continuously evolving; therefore, education for healthcare providers in this field is of paramount importance. ISEMIR's international faculty and world-renowned experts presented the newest information as it relates to the use of magnetic resonance imaging (MRI) and ultrasound (US) at the 8th annual ISEMIR meeting that took place on April 17-18 in Santa Monica, California. Presentations from the meeting can be viewed at www.isemir.org.


Isolation, structure elucidation and in vivo hepatoprotective potential of trans-tetracos-15-enoic acid from Indigofera tinctoria Linn.
The bioassay guided fractionation of the dried aerial part of Indigofera tinctoria Linn. led to the identification of an active fraction labelled as indigotin. On further chemical analysis, a compound isolated from indigotin was identified and characterized as trans-tetracos-15-enoic acid (TCA). The chemical structure of this compound was established on the basis of physical properties and spectral data, including NMR. It afforded significant hepatoprotection against carbon tetrachloride and paracetamol induced hepatotoxicity in experimental models. Silymarin, a well known plant based hepatoprotective agent, and N-acetylcysteine, which has proven efficacy as a replenisher of sulfhydryls, were used for relative efficacy. TCA was found to reverse the altered hepatic parameters in experimental liver damage. In the safety evaluation study the oral LD50 was found to be more than 2000 mg/kg, with no signs of abnormalities or any mortality for the 15 day period of observation after administration of a single dose of drug in mice. The studies revealed significant and concentration dependent hepatoprotective potential of TCA as it reversed the majority of the altered hepatic parameters in experimental liver damage in rats and mice and may be useful in the management of liver disorders.


Fibroblasts Fuel Immune Escape in the Tumor Microenvironment.
Immune escape is central to the persistence of most, if not all, solid tumors and poses a critical obstacle to successful cancer (immuno)therapy. Cancer-associated fibroblasts (CAFs) constitute the most prevalent, yet heterogeneous, component of the tumor stroma, where they 'cool down' the immune microenvironment. The central role played by CAFs, both as a physical barrier and source of immunosuppressive molecules, sets them as a target to enhance immunotherapy of cancer. We outline the current understanding of how CAFs fuel immune escape, as well as their potential clinical applications. Whether these therapeutics really have clinically significant activity remains to be seen, but the outlook is positive.


Refining biome labeling for large-scale microbial community samples: Leveraging neural networks and transfer learning.
Microbiome research has generated an extensive amount of data, resulting in a wealth of publicly accessible samples. Accurate annotation of these samples is crucial for effectively utilizing microbiome data across scientific disciplines. However, a notable challenge arises from the lack of essential annotations, particularly regarding collection location and sample biome information, which significantly hinders environmental microbiome research. In this study, we introduce Meta-Sorter, a novel approach utilizing neural networks and transfer learning, to enhance biome labeling for thousands of microbiome samples in the MGnify database that have incomplete information. Our findings demonstrate that Meta-Sorter achieved a remarkable accuracy rate of 96.7% in classifying samples among the 16,507 lacking detailed biome annotations. Notably, Meta-Sorter provides precise classifications for representative environmental samples that were previously ambiguously labeled as "Marine" in MGnify, thereby elucidating their specific origins in benthic and water column environments. Moreover, Meta-Sorter effectively distinguishes samples derived from human-environment interactions, enabling clear differentiation between environmental and human-related studies. By improving the completeness of biome label information for numerous microbial community samples, our research facilitates more accurate knowledge discovery across diverse disciplines, with particular implications for environmental research.


Molecular analysis of SAR1-related cDNAs from a mouse pituitary cell line.
Vesicular transport between the endoplasmic reticulum (ER) and the Golgi in the yeast Saccharomyces cerevisiae requires a Ras-like, small GTP-binding protein, Sar1p [1-3]. Whether a functional homologue operates in export from the ER in mammalian cells is unknown, nor is it clear if transport in other branches of the secretory pathway requires member(s) of a gene family. In this study, we used a PCR approach to examine the complexity of SAR1-related sequences expressed in mammalian cells that possess multiple secretory pathways. Amplification of cDNA sequences from rodent pituitary cells with primers corresponding to two conserved GTP binding domains of Sar1p yielded several clones with sequences homologous to Sar1 and/or the closely related ADP-ribosylation factor (ARF) family. Of these, only two showed closer homologies to S. cerevisiae Sar1 than members of the ARF family and are designated as mSARa and mSARb. Northern blot analysis shows that mSARa is expressed in most tissues including liver, heart, brain, skeletal muscle and kidney. In contrast, mSARb is preferentially expressed in skeletal muscle and liver. The full-length cDNA of mSARa isolated from a mouse pituitary AtT-20 cDNA library encodes a protein of 198 amino acids, and is 61.6% identical to Sar1p from S. cerevisiae. Thus in contrast to the large rab family of GTP-binding proteins, vesicular transport in mammalian cells appears to be mediated by a relatively small number of Sar1-related proteins.


Simultaneous determination of arabinitol and mannose by gas-liquid chromatography in experimental candidiasis.
A method is described for the simultaneous quantitation of D-arabinitol and D-mannose in serum by gas-liquid chromatography as an aid for the diagnosis of disseminated candidiasis. Both variables were observed as per-O-acetylated aldononitrile derivatives in each chromatographic run of sera from immunosuppressed rabbits experimentally infected with Candida albicans 3181A.


Phase resetting in a model of cardiac Purkinje fiber.
The phase-resetting response of a model of spontaneously active cardiac Purkinje fiber is investigated. The effect on the interbeat interval of injecting a 20-ms duration depolarizing current pulse is studied as a function of the phase in the cycle at which the pulse is delivered. At low current amplitudes, a triphasic response is recorded as the pulse is advanced through the cycle. At intermediate current amplitudes, the response becomes quinquephasic, due to the presence of supernormal excitability. At high current amplitudes, a triphasic response is seen once more. At low stimulus amplitudes, type 1 phase resetting occurs; at medium amplitudes, a type could not be ascribed to the phase resetting because of the presence of effectively all-or-none depolarization; at high amplitudes, type 0 phase resetting occurs. The modeling results closely correspond with published experimental data; in particular type 1 and type 0 phase resetting are seen. Implications for the induction of ventricular arrhythmias are considered.


Decreased proapolipoprotein A-I processing in liver disease: evidence for hepatic participation in proapolipoprotein A-I conversion.
We evaluated the proapolipoprotein A-I (proapo A-I)-converting activity to clarify the pathogenesis of the high proapo A-I/apo A-I ratio in the high-density lipoprotein (HDL) from patients with acute hepatitis and liver cirrhosis. The serum proapo A-I-converting activities were measured using 3H-labeled proapo A-I. 3H-labeled proapo A-I was purified from the media of cultured Hep G2 cells by immunoaffinity chromatography. The serum proapo A-I-converting activities were found to be significantly reduced in the patient with liver cirrhosis (140 +/- 53 dpm/ml per h) or acute hepatitis (140 +/- 48 dpm/ml per h) compared to normal subjects (315 +/- 32 dpm/ml per h). Serum proapo A-I-converting activity has a positive correlation with liver function tests such as serum albumin, choline esterase activity, ICG clearance and inverse correlation with proapo A-I/apo A-I ratio in HDL. These results suggest that the high proapo A-I/apo A-I ratio is due to the decreased proapo A-I-converting activity, and that the liver plays a significant role in the conversion of proapo A-I to apo A-I.


Feedback-controlled negative pressure ventilation in patients with stable severe hypercapnic chronic obstructive pulmonary disease.
In recent studies, the efficacy of intermittent rest of the inspiratory muscles as an option of treating patients with severe chronic obstructive pulmonary disease (COPD) has become questionable. The aim of our study was to analyze the effects of feedback-controlled intermittent negative pressure ventilation (INPV) on stable, but severely hypercapnic COPD patients. 21 clinically stable, hypercapnic patients with severe COPD underwent INPV with chest shells for 3 weeks, 6 h a day. The INPV sessions were optimized by a visual biofeedback system, which enabled control over the decrease in diaphragmatic activity. Respiratory muscle (RM) function parameters, lung function parameters, blood gases and exercise capacity were analyzed. In the end, 19 patients concluded INPV treatment. They had PaO(2) of 56.5 +/- 11.8 mm Hg, PaCO(2) of 50.2+/-2.7 mm Hg (mean +/- SD) and FEV(1) of 27.8 +/- 4.3% predicted before treatment. There was no statistically significant change in lung function parameters, RM function parameters, physical performance and level of dyspnea after 3 weeks of INPV. We conclude that intermittent RM rest induced by INPV can relax inspiratory muscles in most patients with stable severe COPD, but fails to improve RM function and exercise capacity.


The relationship of working memory to the immediate recall of stories following unilateral temporal or frontal lobectomy.
Patients undergoing unilateral temporal lobectomy were tested preoperatively for working memory capacity, sentence comprehension, and rapid-naming ability, and were retested on these same measures 3 to 5 days and 2 weeks postoperatively. Immediate recall of stories was assessed twice only: preoperatively, and 2 weeks postoperatively. The left temporal-lobe group demonstrated transient deficits in working memory capacity, sentence comprehension, and rapid-naming ability over the 2 week postoperative period. No significant group differences in working memory capacity were evident two weeks after operation, even though the story recall of the left temporal-lobe group was impaired. A second experiment, with additional patients, explored the possible contribution of the hippocampus and of the frontal lobes to performance on these same tasks. Extent of hippocampal excision did not affect performance differentially on any task. Story recall was impaired following left temporal lobectomy, whereas left frontal lobectomy impaired sentence comprehension and rapid-naming ability. These results indicate a double dissociation of verbal deficits 2 weeks after left temporal- and left frontal-lobe excisions.


Impact of different immunosuppressive regimens on the health-related quality of life following orthotopic liver transplantation.
The influence of immunosuppression on the recipients' quality of life (QoL) is of major importance after OLT and has not yet been evaluated. The impact of different immunosuppression regimens after OLT was evaluated in 275 patients using the Short Form 36 (SF-36) survey. The following immunosuppressive strategies were compared: (a) CNI, (b) mTOR inhibitors, and (c) mTOR combined with CNI. All regimens were prescribed alone (mono) or in combination (+) with prednisolone and/or mycophenolate mofetil (MMF). Highest scores were evident in patients in the mTOR+ group. There were significantly higher values for general health perceptions (GH, p = 0.049), vitality (VIT, p = 0.020), and physical component summary (PCS, p = 0.041) when compared to CNImono and for GH (p = 0.042) and VIT (p = 0.043), when compared to mTORmono. Early conversion to mTOR inhibitors (<two months after OLT) was associated with higher values for 7 of 10 scales, when compared to a late conversion (>two months after OLT), with a statistically significant improvement for the dimension role-emotional (RE, p = 0.027). mTOR inhibitor-based regimens appear to have beneficial effects on QoL after OLT, especially after an early conversion.


[Expression of plasminogen activator inhibitor-1 and tissue plasminogen activator in the kidney of KKAy mice with type 2 diabetes].
To study the role of plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (tPA) in the accumulation of extracellular matrix (ECM) in the kidney of KKAy mice with type 2 diabetes. KKAy mice, a type 2 diabetic animal model, and C57BL-J mice were sacrificed at 16, 20, and 24 weeks of age, respectively. The local expression of renal laminin was analyzed with immunohistochemistry. Chromogenic substance was used to show the activity of PAI-1. The mRNA expression of tPA was determined by RT-PCR. The mRNA expression of PAI-1 was measured by reverse transcription-polymerase chain reaction (RT-PCR). Laminnin expression was significantly increased in all age groups of KKAy mice. The tPA mRNA was significantly lower than that in C57BL mice, especially at the age of 16w (only 47%). Otherwise the PAI-1 mRNA expression was remarkably up-regulated than that in C57BL mice. In type 2 diabetes KKAy mice, the accumulation of ECM may be associated with the abnormal expression of PAI-1/tPA mRNA.


Perfluoroalkylated substances (PFASs) and legacy persistent organic pollutants (POPs) in halibut and shrimp from coastal areas in the far north of Norway: Small survey of important dietary foodstuffs for coastal communities.
Halibut (Hippoglossus hippoglossus) and shrimps (Pandalus borealis) are regular foodstuffs for communities in northern Norway and important species for the coastal fishing industry. This is the first study to present a comprehensive overview of the contaminant status of these species, with emphasis on unregulated perfluoroalkylated substances (PFAS). The contaminant concentrations were low and within tolerable levels for human dietary exposure. Median Σpolychlorinated biphenyls (PCB) were 4.9 and 2.5ng/g ww for halibut and unpeeled shrimps, respectively. Concentrations of perfluorooctane sulfonate (PFOS) - the most abundant PFASs - were 0.9 and 2.7ng/g ww in halibut and shrimp, respectively. The halibut fillets were dominated by PCBs, which contributed to 50% of the total POPs load, followed by ΣDDTs; 26% and PFASs (18%), whereas shrimps were dominated by PFASs (74%). ΣPBDEs (polybrominated diphenyl ethers) contributed to 1-4% of the total POP load. Local sources are not contributing significantly to the contaminant burden in these species.


Neuropathology in mice expressing mouse alpha-synuclein.
α-Synuclein (αSN) in human is tightly linked both neuropathologically and genetically to Parkinson's disease (PD) and related disorders. Disease-causing properties in vivo of the wildtype mouse ortholog (mαSN), which carries a threonine at position 53 like the A53T human mutant version that is genetically linked to PD, were never reported. To this end we generated mouse lines that express mαSN in central neurons at levels reaching up to six-fold compared to endogenous mαSN. Unlike transgenic mice expressing human wildtype or mutant forms of αSN, these mαSN transgenic mice showed pronounced ubiquitin immunopathology in spinal cord and brainstem. Isoelectric separation of mαSN species revealed multiple isoforms including two Ser129-phosphorylated species in the most severely affected brain regions. Neuronal Ser129-phosphorylated αSN occurred in granular and small fibrillar aggregates and pathological staining patterns in neurites occasionally revealed a striking ladder of small alternating segments staining either for Ser129-phosphorylated αSN or ubiquitin but not both. Axonal degeneration in long white matter tracts of the spinal cord, with breakdown of myelin sheaths and degeneration of neuromuscular junctions with loss of integrity of the presynaptic neurofilament network in mαSN transgenic mice, was similar to what we have reported for mice expressing human αSN wildtype or mutant forms. In hippocampal neurons, the mαSN protein accumulated and was phosphorylated but these neurons showed no ubiquitin immunopathology. In contrast to the early-onset motor abnormalities and muscle weakness observed in mice expressing human αSN, mαSN transgenic mice displayed only end-stage phenotypic alterations that manifested alongside with neuropathology. Altogether these findings show that increased levels of wildtype mαSN does not induce early-onset behavior changes, but drives end-stage pathophysiological changes in murine neurons that are strikingly similar to those evoked by expression of human wildtype or mutant forms.


Molecular and Structural Characterization of Lenalidomide-Mediated Sequestration of eIF3i.
Lenalidomide is a ligand of the E3 ligase substrate adapter cereblon (CRBN) that achieves its clinical effects in part by the promotion of substrate recruitment and degradation. In contrast to prior targets, eIF3i is recruited but not degraded upon complex formation with lenalidomide and CRBN, although the structural details and mechanistic outcomes of this interaction are unresolved. Here, we characterize the structural basis and mechanistic outcomes of lenalidomide-induced sequestration of eIF3i from the eIF3 complex. Identification of the binding interface on eIF3i by a covalent lenalidomide probe and mass spectrometry rationalizes the sequestration event. We further connect eIF3i and CRBN to lenalidomide-driven effects on angiogenic markers, Akt1 phosphorylation, and associated antiangiogenesis phenotypes. Finally, we find that eIF3i sequestration is observed in MM.1S and MOLM13 cells after the degradation of other substrates, such as IKZF1. The defined binding interface elucidated by chemical proteomics and the observation of eIF3i sequestration as a lenalidomide function open future directions in designing new chemical adapters for protein sequestration as a strategy to selectively control protein functions.


Use of chromophoric ligands to visually screen co-crystals of putative protein-nucleic acid complexes.
Distinguishing between crystals of protein-nucleic acid complexes and those containing protein alone is a common problem in structural studies of protein-nucleic acid interactions. Currently, there are several methods available for detecting nucleic acid in crystals, including gel electrophoresis, SYBR Gold fluorescence dye staining, and methyl violet staining. However, they require either that the crystals be sacrificed or access to a fluorescence microscope. In this protocol, we describe an approach that allows direct visualization of either the presence or absence of oligonucleotides in crystals grown from solutions containing both protein and nucleic acid--labeling with the Cy5 dye. In addition to offering the advantage of being able to distinguish between crystals of complex and protein alone with the naked eye or a light microscope, crystals of covalently Cy5-labeled DNA can be directly used for X-ray diffraction data collection.


Higher testosterone is associated with open-angle glaucoma in women: a genetic predisposition?
Testosterone may be a possible modifiable risk factor for open-angle glaucoma (OAG) and intraocular pressure (IOP), but evidence has been scarce and conflicting. In this study we evaluated the association of testosterone and its genetic predisposition with incident (i) OAG, IOP, retinal nerve fiber layer (RNFL), and ganglion cell-inner plexiform layer (GCL +). Participants aged 45-100 years were derived from the prospective, population-based Rotterdam Study. Ophthalmic examinations and serum testosterone measurements (including bioavailable and free testosterone) were performed from 1991 onwards. Follow-up took place every 4-5 years. A total of 187 out of 7898 participants were diagnosed with incident (i) OAG during follow-up. Genotyping was performed in 165 glaucoma cases and 6708 controls. We calculated sex-specific weighted genetic risk scores (GRS) for total and bioavailable testosterone. Associations with iOAG were analyzed using multivariable logistic regression. Associations with IOP, RNFL, and GCL + were analyzed with multivariable linear regression. Analyses were stratified on sex and adjusted for at least age, body mass index, and follow-up duration. In men, testosterone was not associated with iOAG. However, the GRS for higher total testosterone was associated with an increased iOAG risk (odds ratio [OR] with 95% confidence interval [95% CI]: 2.48 [1.18; 5.22], per unit). In women, higher values of bioavailable testosterone (2.05 [1.00; 4.18] per nmol/L) and free testosterone (1.79 [1.00; 3.20] per ng/dL) were significantly associated with increased risk of iOAG. Moreover, the GRS for higher bioavailable testosterone was associated with an increased iOAG risk (2.48 [1.09; 5.65], per unit). Higher bioavailable and free testosterone were adversely associated with IOP (0.58 [0.05; 1.10] per nmol/L and 0.47 [0.04; 0.90] per ng/dL). Higher total testosterone was inversely associated with peripapillary RNFL and GCL + (Beta [95% CI]: - 3.54 [- 7.02; - 0.06] per nmol/L and - 2.18 [- 4.11; - 0.25] per nmol/L, respectively). In women, higher testosterone levels increased the risk of iOAG. Both IOP-dependent and IOP-independent mechanisms may underlie this association. Managing testosterone levels may be particularly relevant for the prevention of neurodegeneration in the eye. Future research should confirm these findings.


Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines.
We previously reported that blood acetylcholine (ACh) mainly originates from T-lymphocytes and that muscarinic (Ms) ACh receptor mRNA is expressed in both T- and B-lymphocytes. In the present study, we used confocal laser scanning microscopy and fluo-3, a calcium-sensitive indicator, to investigate the effects of Ms-ACh receptor agonists on the intracellular free Ca2+ concentration ([Ca2+]i) in single cells from human T-cell (CEM) and B-cell (Daudi) lines, which we used as models of lymphocytes. In both cell lines, stimulation of Ms-ACh receptors with ACh (0.1-100 microM), bethanechol (100 microM), car-bachol (100 microM) or oxotremorine-M (Oxo-M; 0.1-100 microM) induced [Ca2+]i-dependent increases in fluo-3 fluorescence, which in the presence of extracellular Ca2+ were followed by oscillations in [Ca2+]i that persisted for at least 10 min. All effects were completely blocked by atropine (1 microM), an Ms-ACh receptor antagonist. In both cell lines Oxo-M (100 microM) up-regulated expression of c-fos mRNA in an extracellular Ca2+-dependent manner. Again, the effect was blocked by 1 microM atropine. These results provide the first evidence that stimulation of Ms-ACh receptors induces Ca2+ oscillations and up-regulates c-fos gene expression in T- and B-lymphocytes, which is consistent with the notion that ACh released from T-lymphocytes triggers nuclear signaling via Ms-ACh receptors.


Effect of CYP2D6 pharmacogenetic phenotype and phenoconversion on serum concentrations of antidepressants and antipsychotics: a retrospective cohort study.
Pharmacogenetics (PGx), especially in regard to CYP2D6, is gaining more importance in routine clinical settings. Including phenoconversion effects (PC) in result interpretation could maximize its potential benefits. However, studies on genetics of pharmacokinetic genes including the functional enzyme status are lacking. The retrospective analyses of clinical routine data aimed to investigating how the CYP2D6 functional enzyme status affects serum concentrations and metabolite-to-parent ratios of seven common psychotropic drugs and allows an evaluation of the relevance of this information for patient care. Two patient cohorts (total n = 316; 44.2 ± 15.4 years) were investigated for the CYP2D6 functional enzyme status and its associations with drug exposure and metabolism of venlafaxine, amitriptyline, mirtazapine, sertraline, escitalopram, risperidone and quetiapine. We found an increase in intermediate and poor metabolizers, as well as a decrease in normal metabolizers of CYP2D6 when including PC. Moreover, we found associations between amitriptyline exposure with the phenoconversion-corrected activity score of CYP2D6 (Spearman correlation; p = 0.03), and risperidone exposure with CYP2D6 functional enzyme status (Kruskal-Wallis test; p = 0.01), as well as between metabolite-to-parent ratio of venlafaxine and risperidone with CYP2D6 functional enzyme status (Kruskal-Wallis test; p < 0.001; p = 0.05). The data stress the relevance of PC-informed PGx in psychopharmacological treatment and suggest that PC should be included in PGx result interpretation when PGx is implemented in routine clinical care, especially before initiating amitriptyline- or risperidone-treatment, to start with a dose adequate to the respective CYP2D6 functional enzyme status. Moreover, PGx and therapeutic drug monitoring should be used complementary but not alternatively.


Efficient removal of aqueous Cr(VI) with ferrous sulfide/N-doped biochar composites: Facile, in-situ preparation and Cr(VI) uptake performance and mechanism.
FeS nanoparticles loaded on nitrogen-doped biochar (FeS/BNC) were fabricated by pyrolyzing coffee husks pretreated with Mohr's salt. The nitrogen doping and FeS loading of biochar are simultaneously achieved in one-pot pyrolysis. The elemental analysis, SEM, TEM, XRD, XPS, Raman, FTIR and N2 adsorption-desorption technologies were used to characterize the composition and structure of FeS/NBC. The appraisement for removing aqueous Cr(VI) testified that FeS/NBC offered a synergistic scavenging effect of Cr(VI) by FeS and NBC. The effect of crucial experimental conditions (FeS/NBC dosage, foreign ions, initial pH and concentration of Cr(VI) solution) were investigated. The Cr(VI) removal capacity was as high as 211.3 ± 26 mg g-1 under the optimized condition. The practicability of FeS/NBC was examined by using simulated actual samples from tap water and lake water. The mechanism examination showed that surface adsorption/reduction and solution reduction were implicated in the removal of Cr(VI). The current work introduces a novel FeS/NBC composite prepared by an in situ pyrolysis method with excellent potential for chromium pollution remediation.


Activite succino-deshydrogenasique des microsomes et mode d'incorporation du succinate 2,3-(14)C dans les acides gras des microsomes de foie de rat in vitro.
The incorporations of 2,3-(14)C-succinate 2-(14)C-acetate into fatty acids of different cellular fractions of rat liver were studied. Acetate was incorporated mainly into supernatant and succinate into microsomal fatty acids. Mitochondria only could intensively decarboxylate pyruvate. Avidine inhibited fatty acid synthesis from succinate mainly in the supernatant. It is suggested that succinate is an important physiological precursor of fatty acids in the liver and that an active succino-dehydrogenase is present in microsomes.


Expression of IL-1β in rhesus EAE and MS lesions is mainly induced in the CNS itself.
Interleukin (IL)-1β is a pro-inflammatory cytokine that plays a role in the pathogenesis of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), the animal model for MS. Yet, detailed studies on IL-1β expression in different stages of MS lesion development and a comparison of IL-1β expression in MS and EAE are lacking. Here, we performed an extensive characterization of IL-1β expression in brain tissue of MS patients, which included different MS lesion types, and in brain tissue of rhesus macaques with EAE. In rhesus EAE brain tissue, we observed prominent IL-1β staining in MHC class II(+) cells within perivascular infiltrates and at the edges of large demyelinating lesions. Surprisingly, staining was localized to resident microglia or differentiated macrophages rather than to infiltrating monocytes, suggesting that IL-1β expression is induced within the central nervous system (CNS). By contrast, IL-1β staining in MS brain tissue was much less pronounced. Staining was found in the parenchyma of active and chronic active MS lesions and in nodules of MHC class II(+) microglia in otherwise normal appearing white matter. IL-1β expression was detected in a minority of the nodules only, which could not be distinguished by the expression of pro- and anti-inflammatory markers. These nodules were exclusively found in MS, and it remains to be determined whether IL-1β(+) nodules are destined to progress into active lesions or whether they merely reflect a transient response to cellular stress. Although the exact localization and relative intensity of IL-1β expression in EAE and MS is different, the staining pattern in both neuroinflammatory disorders is most consistent with the idea that the expression of IL-1β during lesion development is induced in the tissue rather than in the periphery.


The effect of PARO robotic seals for hospitalized patients with dementia: A feasibility study.
Robotic seals have been studied in long-term care settings; though, no studies of patients with dementia in the acute care setting have been reported. The purpose of this study was to evaluate the feasibility of PARO interventions for hospitalized patients with dementia, determine physiological effects and describe social-affective interactions. Using a prospective descriptive design with pre-post PARO intervention physiological measurements, we studied 55 participants who received up to five 15-min PARO interventions. The PARO was favorably accepted for 212 (95%) of the 223 PARO interventions. Differences in pre- and post-physiological measures for mean arterial pressure, pulse, respiration, oxygenation, stress, and pain levels were evaluated using Wilcoxon Signed Rank test with statistically significant pre and post differences (p=<0.05); however, the differences were not clinically significant. Participants (95%) demonstrated beneficial PARO interactions with the most frequent interactions being speaking and petting. The PARO shows promise for enhancing social and affective responses for hospitalized patients with dementia.


Conditional gene expression systems to study herpesvirus biology in vivo.
Cytomegalovirus (CMV), a prototypic beta-herpesvirus, is an important human pathogen causing protean clinical manifestations in immature and immunocompromised patients. Mechanisms of infection can be studied in a mouse model. Mouse cytomegalovirus (MCMV) resembles in pathogenesis its human counterpart in many ways. Although MCMV infection is studied extensively on the level of organs, the contribution of specific cell types to viral replication in vivo is still elusive. Here we describe our approach based on the the Cre/loxP-system to investigate MCMV infection at the level of cell types in vivo. Using bacterial artificial chromosome (BAC)-technology, we created an MCMV virus containing an enhanced green fluorescent protein (egfp) reporter-gene which is not expressed due to a 'Stop' cassette flanked by two loxP-sites between promoter and coding sequence. Infection of cre-transgenic mice with this reporter virus results in the deletion of the 'Stop' cassette and expression of EGFP in a cell type-specific manner. Using this conditional gene expression system we are able to quantify viral productivity in specific cell types and to determine their contribution to viral dissemination in vivo. Furthermore, the deletion of viral genes can be used to screen for cell type-specificity of viral gene functions. Hence, conditional MCMV mutants allow the study of herpesvirus biology on the level of cell types in vivo.


Purification of molecular machines and nanomotors using phage-derived monoclonal antibody fragments.
Molecular machines and nanomotors are sophisticated biological assemblies that convert potential energy stored either in transmembrane ion gradients or in ATP into kinetic energy. Studying these highly dynamic biological devices by X-ray crystallography is challenging, as they are difficult to produce, purify, and crystallize. Phage display technology allows us to put a handle on these molecules in the form of highly specific antibody fragments that can also stabilize conformations and allow versatile labelling for electron microscopy, immunohistochemistry, and biophysics experiments.Here, we describe a widely applicable protocol for selecting high-affinity monoclonal antibody fragments against a complex molecular machine, the A-type ATPase from T. thermophilus that allows fast and simple purification of this transmembrane rotary motor from its wild-type source. The approach can be readily extended to other integral membrane proteins and protein complexes as well as to soluble molecular machines and nanomotors.


Decellularized retinal matrix: Natural platforms for human retinal progenitor cell culture.
Tissue decellularization strategies have enabled engineering of scaffolds that preserve native extracellular matrix (ECM) structure and composition. In this study, we developed decellularized retina (decell-retina) thin films. We hypothesized that these films, mimicking the retina niche, would promote human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. Retinas isolated from bovine eyes were decellularized using 1% w/v sodium dodecyl sulfate (SDS) and pepsin digested. The resulting decell-retina was biochemically assayed for composition and cast dried to develop thin films. Attachment, viability, morphology, proliferation and gene expression of hRPC cultured on the films were studied in vitro. Biochemical analyses of decell-retina compared to native retina indicated the bulk of DNA (94%) was removed, while the majority of sulfated GAGs (55%), collagen (83%), hyaluronic acid (87%), and key growth factors were retained. The decell-retina films supported hRPC attachment and growth, with cell number increasing 1.5-fold over a week. RT-PCR analysis revealed hRPC expression of rhodopsin, rod outer membrane, neural retina-specific leucine zipper neural and cone-rod homeobox gene on decell-retina films, indicating photoreceptor development. In conclusion, novel decell-retina films show promise as potential substrates for culture and/or transplantation of retinal progenitor cells to treat retinal degenerative disorders. In this study, we report the development of a novel biomaterial, based on decellularized retina (decell-retina) that mimics the retina niche and promotes human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. We estimated, for the first time, the amounts of collagen I, GAGs and HA present in native retina, as well as the decell-retina. We demonstrated that retinas can be decellularized using ionic detergents and can be processed into mechanically stable thin films, which can act as substrates for culturing hRPCs. Rhodopsin, ROM1, NRL and CRX gene expression on the decell-retina films indicated photoreceptor development from RPCs. These results support the potential of decell-retina as a cell delivery platform to treat and manage retinal degenerative disease like AMD.


Histone H3 lysine 4 methylation disrupts binding of nucleosome remodeling and deacetylase (NuRD) repressor complex.
Histone N-terminal tails are post-translationally modified in many ways. At lysine residues, histones can be either acetylated or methylated. Both modifications lead to the binding of specific proteins; bromodomain proteins, such as GCN5, bind acetyl lysines and the chromodomain protein, HP1, binds methyl lysine 9 of histone H3. Here we show that the previously characterized transcriptional repressor complex NuRD (nucleosome remodeling and deacetylase) binds to the histone H3 N-terminal tail and that methylation at lysine 4, but not lysine 9, prevents binding. Given that lysine 4 methylation is found at sites of active transcription, these results suggest that a function of lysine 4 methylation is to disrupt the association of histones with a repressor complex.


[Allergy to cow's milk].
Cow's milk allergy is a disease of infancy and usually appears in the first few months of life. The evaluation of infants for possible cow's milk allergy is one of the more common problems encountered by pediatricians. Unfortunately, both over and under diagnosis is frequently seen. In this review the pathophysiology, the clinical manifestations, the diagnosis and management of infants with cow's milk allergy as well as the prevention of the condition in predisposed infants are discussed.


[Intrathoracic injuries].
Severe intrathoracic injuries are uncommon but immediately life-threatening. These injuries are mostly associated with polytrauma. After stabilization of polytraumatized patients imaging is a prerequisite for treatment and operation planning. The assessment warrants an interdisciplinary approach primarily between the specialties of anesthesia, trauma surgery and thoracic surgery and further specialties should be involved depending on the injury pattern. This article gives an overview about the current management of the most important intrathoracic injuries.


Behavioural analysis and susceptibility to CNS injury of four inbred strains of mice.
Interpretation of data from gene targeting studies can be confounded by the inherent traits of the background inbred strains used in the generation of transgenic and null mutant mice. We have therefore compared the behaviour and response to CNS injury of four inbred strains commonly used in molecular genetic studies to produce models of neurological disease. Adult, male 129/Ola, BALB/c, C57BL/6 and FVB/N mice (2-4 months) were initially subjected to behavioural tests that comprised a neurological examination, determination of motor function and cognitive testing in the Morris water maze. Also the response to CNS injury following an acute kainic acid (KA) challenge (30 mg kg-1, i.p.) was determined. The 129/Ola and BALB/c strains showed significant motor deficits when compared with the C57BL/6 and FVB/N strains. In contrast, only the FVB/N strain showed evidence of apparent cognitive impairments in the water maze as evidenced by increased pathlengths to locate the escape platforms and impaired performance in a probe trial. In addition, the FVB/N strain showed the most severe seizure response and mortality rate (62%) following administration of KA (30 mg kg-1, i.p.). These behavioural changes were also associated with a greater degree of cell body and synaptophysin loss in the pyramidal CA3 hippocampal cell layer and astrogliosis 72-h post-dose. These data suggest that the FVB/N strain may not be the most suitable background strain for the development of new transgenic mice for the study of genes implicated in the learning and memory process.


[About the certainty of study results. What p-value and confidence intervalls tell us on it].
In clinical trials data of individuals are collected, who are subjected to different treatments under controlled conditions. These studies investigate differences between interventions, and their results--provide the basis for clinical decision making in individuals or for policy decisions concerning entire patient populations. This requires reliable results. How confident a result of the study actually is, is illustrated by using various statistical parameters: p-value or confidence interval. What these parameters are and how to interpret them, is the objective of this article.