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Expression of ceramide anabolizing and catabolizing enzymes in MIN and CIN colorectal cells upon DL-PDMP treatment.

UGCG and SGMS1 knockdown efficiencies with different shRNA and siRNA.

DL-PDMP inhibits the proliferation of myeloid leukemia cell lines.

DL-PDMP does not affect proliferation of MEFs and colorectal cancer cells.

DL-PDMP treatment does not cause weight loss in mice.

Aneuploidy levels in Bub1b H/H and Cdc20 AAA/AAA MEFs.

Aneuploidy, a hallmark of cancer cells, poses an appealing opportunity for cancer treatment and prevention strategies. Using a cell-based screen to identify small molecules that could selectively kill aneuploid cells, we identified the compound N-[2-hydroxy-1-(4-morpholinylmethyl)-2-phenylethyl]-decanamide monohydrochloride (DL-PDMP), an antagonist of UDP-glucose ceramide glucosyltransferase. DL-PDMP selectively inhibited proliferation of aneuploid primary mouse embryonic fibroblasts and aneuploid colorectal cancer cells. Its selective cytotoxic effects were based on further accentuating the elevated levels of ceramide, which characterize aneuploid cells, leading to increased apoptosis. We observed that DL-PDMP could also enhance the cytotoxic effects of paclitaxel, a standard-of-care chemotherapeutic agent that causes aneuploidy, in human colon cancer and mouse lymphoma cells. Our results offer pharmacologic evidence that the aneuploid state in cancer cells can be targeted selectively for therapeutic purposes, or for reducing the toxicity of taxane-based drug regimens. Cancer Res; 77(19); 5272–86. ©2017 AACR.

Sphingolipid composition of aneuploid and euploid cell lines upon DL-PDMP treatment.

Table S1: Summary of compound effect in aneuploid Ts13 MEF compared to euploid wild-type MEF. Table S2: shRNA and siRNA sequences used in this study. Table S3: Primers used for quantitative Real-Time PCR.

Single cell biology has the potential to elucidate many critical biological processes and diseases, from development and regeneration to cancer. Single cell analyses are uncovering the molecular diversity of cells, revealing a clearer picture of the variation among and between different cell types. New techniques are beginning to unravel how differences in cell state-transcriptional, epigenetic, and other characteristics-can lead to different cell fates among genetically identical cells, which underlies complex processes such as embryonic development, drug resistance, response to injury, and cellular reprogramming. Single cell technologies also pose significant challenges relating to processing and analyzing vast amounts of data collected. To realize the potential of single cell technologies, new computational approaches are needed. On March 17-19, 2021, experts in single cell biology met virtually for the Keystone eSymposium "Single Cell Biology" to discuss advances both in single cell applications and technologies.

Cobalt germanides have been widely studied as semiconductor contact materials, but recent theoretical studies suggest that they may also be excellent catalysts for methane steam reforming with stabilities and activities comparable to more expensive noble metal catalysts. We have sputter deposited CoGe alloy films and characterized their structure and morphology after post-deposition annealing in high vacuum up to 1000 °C. We used X-ray photoelectron spectroscopy to study the initial oxidation of amorphous and crystalline CoGe alloy surfaces under low pressures of O2 and H2O. The oxidation rate in O2 was found to be faster for an amorphous CoGe surface compared to a crystalline surface. We also found that there was little difference in the oxidation rate in H2O for either amorphous or crystalline surfaces. During O2 oxidation, the crystalline surface preferentially forms GeO and the amorphous surface preferentially forms GeO2. We have also observed preferential oxidation of Ge in the CoGe thin films. During temperature programmed desorption studies, we found that GeO desorption begins near 350 °C and that GeO2 decomposes to GeO and desorbs near 700 °C. More studies of CoGe catalysts are warranted, however GeO desorption may be a concern under reaction conditions when the film is subjected to an oxidizing environment.

SUMMARYThe blood-brain barrier (BBB) is critical for protecting the brain and maintaining neuronal homeostasis. Although the BBB is a unique feature of the central nervous system (CNS) vasculature, not all brain regions have the same degree of impermeability. Differences in BBB permeability are important for controlling the local extracellular environment of specific brain regions to regulate the function and plasticity of particular neural circuits. However, how BBB heterogeneity occurs is poorly understood. Here, we demonstrate how regional specialization of the BBB is achieved. With unbiased cell profiling in small, defined brain regions, we compare the median eminence, which has a naturally leaky BBB, with the cortex, which has an impermeable BBB. We identify hundreds of molecular differences in endothelial cells (ECs) and demonstrate the existence of differences in perivascular astrocytes and pericytes in these regions, finding 3 previously unknown subtypes of astrocytes and several key differences in pericytes. By serial electron microscopy reconstruction and a novel, aqueous-based tissue clearing imaging method, we further reveal previously unknown anatomical specializations of these perivascular cells and their unique physical interactions with neighboring ECs. Finally, we identify ligand-receptor pairs between ECs and perivascular cells that may regulate regional BBB integrity in ECs. Using a bioinformatic approach we identified 26 and 26 ligand-receptor pairs underlying EC-pericyte and EC-astrocyte interactions, respectively. Our results demonstrate that differences in ECs, together with region-specific physical and molecular interactions with local perivascular cells, contribute to BBB functional heterogeneity. These regional cell inventories serve as a platform for further investigation of the dynamic and heterogeneous nature of the BBB in other brain regions. Identification of local BBB specializations provides insight into the function of different brain regions and will permit the development of region-specific drug delivery in the CNS.

The blood–brain barrier (BBB) restricts free access of molecules between the blood and the brain and is essential for regulating the neural microenvironment. Here, we describe how the BBB was initially characterized and how the current field evaluates barrier properties. We next detail the cellular nature of the BBB and discuss both the conservation and variation of BBB function across taxa. Finally, we examine our current understanding of mouse and zebrafish model systems, as we expect that comparison of the BBB across organisms will provide insight into the human BBB under normal physiological conditions and in neurological diseases.

A bounding risk assessment is presented that evaluates possible human health risk from a hypothetical scenario involving a 10,000-gallon release of flowback water from horizontal fracturing of Marcellus Shale. The water is assumed to be spilled on the ground, infiltrates into groundwater that is a source of drinking water, and an adult and child located downgradient drink the groundwater. Key uncertainties in estimating risk are given explicit quantitative treatment using Monte Carlo analysis. Chemicals that contribute significantly to estimated health risks are identified, as are key uncertainties and variables to which risk estimates are sensitive. The results show that hypothetical exposure via drinking water impacted by chemicals in Marcellus Shale flowback water, assumed to be spilled onto the ground surface, results in predicted bounds between 10-10 and 10-6 (for both adult and child receptors) for excess lifetime cancer risk. Cumulative hazard indices (HICUMULATIVE ) resulting from these hypothetical exposures have predicted bounds (5th to 95th percentile) between 0.02 and 35 for assumed adult receptors and 0.1 and 146 for assumed child receptors. Predicted health risks are dominated by noncancer endpoints related to ingestion of barium and lithium in impacted groundwater. Hazard indices above unity are largely related to exposure to lithium. Salinity taste thresholds are likely to be exceeded before drinking water exposures result in adverse health effects. The findings provide focus for policy discussions concerning flowback water risk management. They also indicate ways to improve the ability to estimate health risks from drinking water impacted by a flowback water spill (i.e., reducing uncertainty).

Only thin-film technology can satisfy the requirements of high spatial selectivity at high-channel-count electrode array designs by simultaneously good conformability to the targeted tissue through mechanical flexibility enriching future applications of functional neural stimulation. However, caused by the high impact of the microstructure on the mechanical and electrochemical film properties, varying fabrication processes of the same thin-film makes the difference between acute and chronic long-term stable electrodes. The influence of standard clinical electrical pulsing on flexible polyimide-based thin-film platinum electrodes for neuroprostheses, either sputter deposited or evaporated, and different diameters was assessed and compared. The electrochemical and morphological analysis showed a higher corrosion susceptibility and electrochemical degradation for the sputter deposited platinum electrodes with even total failures of smaller diameters. In contrast, the evaporated thin-films provided itself as more stable and reliable metallization with also smaller electrodes keeping their film integrity intact over the experimental period, -appearing to be the preferable material for improving thin-film electrodes' longevity.

The high complexity of the biological response to implanted materials builds a serious barrier against implanted recording and stimulation electrode arrays to succeed in clinically relevant chronic studies. Some of the cell and molecular interactions and their contribution to inflammation and device failure are still unclear. The interrelated mechanisms leading to tissue damage and electrode array failure during simultaneous faradaic, electrochemical reactions and biological response under electrical stimulation are not understood sufficiently. One variable, with which inflammatory and electrode surface processes can be analyzed and assessed, is the pH change in the immediate environment of the material-tissue interface. Here, the greatest challenges are in the biocompatibility and in-vivo long-term stability of selected sensor materials, the measurement of small transient pH oscillations and positioning of the sensor at a defined and nearest possible distance in the micrometer range, to the site of activity without the pH sensing being affected by the material- issue interactions itself. This work represents the in-situ measurement of local and transient pH changes at apulsed electrode with an embedded in-vivo compatible pH sensor and therein differentiating from current approaches of pH sensing during electrical stimulation.

The aim of the current study was to evaluate the effectiveness of intraoperative neuromonitoring (INM) as an adjunct in performing tarsal tunnel decompression surgery. We reviewed 38 patients who met inclusion criteria. INM was used to measure the voltage of the abductor hallucis and digiti quinti muscles both before and after decompression. Observed changes intraoperatively were acute and within minutes of the decompression performed by the surgeon. Patient outcomes were ascertained from clinical findings and classified as excellent, fair, or poor. Patient outcomes and the voltage change were measured and assessed for association, and statistically significant differences were found between outcome groups. Of the 38 patients, 29 (76%) had excellent outcomes, with a mean change in microvolts of 2088.28 ± 1172.44 (684%) (p = .0004) and 2173.24 ± 1228.39 (742%) (p = .0014) for abductor hallucis and abductor digiti quinti, respectively. The study supports INM as a useful adjunct in performing tarsal tunnel decompression.

Aneuploidy, a hallmark of cancer cells, poses an appealing opportunity for cancer treatment and prevention strategies. Using a cell-based screen to identify small molecules that could selectively kill aneuploid cells, we identified the compound N-[2hydroxy-1-(4-morpholinylmethyl)-2-phenylethyl]-decanamide monohydrochloride (DL-PDMP), an antagonist of UDP-glucose ceramide glucosyltransferase. DL-PDMP selectively inhibited proliferation of aneuploid primary mouse embryonic fibroblasts and aneuploid colorectal cancer cells. Its selective cytotoxic effects were based on further accentuating the elevated levels of ceramide, which characterize aneuploid cells, leading to increased apoptosis. We observed that DL-PDMP could also enhance the cytotoxic effects of paclitaxel, a standard-of-care chemotherapeutic agent that causes aneuploidy, in human colon cancer and mouse lymphoma cells. Our results offer pharmacologic evidence that the aneuploid state in cancer cells can be targeted selectively for therapeutic purposes, or for reducing the toxicity of taxane-based drug regimens., National Cancer Institute (U.S.) (Grant P30-CA14051)

Miniaturization of electrodes is a prerequisite of selective and targeted interaction with single neurons, enabling more applications in the continuously growing field of neuroprostheses. Miniaturization in all three dimensions of the electrical contact sites should maintain or increase longevity and electrical functionality. The thin-film metallization of the electrode site, which is only a couple of hundreds of nanometers thick, has to withstand high chemical load through the corrosive environment in the body and the electrochemical processes during electrical stimulation in vivo. Platinum (Pt), which is known to be chemically inert and mechanical stable as bulk material shows a lack of chemical and mechanical integrity applied in thin-film microelectrodes. In our study we investigated failure mechanisms of thin-film Pt electrodes under conditions of electrode aging and electrical stimulation in different physiological media. To understand and eventually overcome stability loss, we investigated the intrinsic structural stress and deformations that arose from mechanical loading through chemical impact and electrical stimulation using optical microscopy and white-light interferometry. Electrochemical measurements indicated oxidation and surface roughening as two of the degradation processes in thin-film electrodes. From the results presumptions about the underlying microstructural changes were made.

We investigate the compact integration of passive capacitive filter units embedded in low-temperature cofired ceramics for active implantable medical devices. The goal lies in avoiding volume consumption by large passive components and to achieve a miniaturized integration of filters for the reduction of induced radio frequency currents and voltages during magnetic resonance imaging. Embedding such filter units inside the substrates carrying the electronics can be a promising solution for implants to allow higher channel densities with only a marginal increase in implant's size. By combining laser ablation and screen printing manufacturing technologies, we investigated a fabrication process for multilevel ceramic capacitors (MLCCs) in low temperature co-fired ceramic (LTCC) substrates resulting in mean capacitances of 220 pF, (10 × 0.4 × 0.62 mm3) with a dissipation factor of 0.42% and a breakdown voltage of 500 V. Dissolution of the material during accelerated aging tests however raises concerns over its biostability and biocompatibility during chronic implantation and needs further testing.

A bounding risk assessment is presented that evaluates possible human health risk from a hypothetical scenario involving a 10,000-gallon release of flowback water from horizontal fracturing of Marcellus Shale. The water is assumed to be spilled on the ground, infiltrates into groundwater that is a source of drinking water, and an adult and child located downgradient drink the groundwater. Key uncertainties in estimating risk are given explicit quantitative treatment using Monte Carlo analysis. Chemicals that contribute significantly to estimated health risks are identified, as are key uncertainties and variables to which risk estimates are sensitive. The results show that hypothetical exposure via drinking water impacted by chemicals in Marcellus Shale flowback water, assumed to be spilled onto the ground surface, results in predicted bounds between 10

A method for determining a safety range for non-cancer risks is proposed, similar in concept to the range used for cancer in the management of waste sites. This safety range brings transparency to the chemical specific Reference Dose or Concentration by replacing their “order of magnitude” definitions with a scientifically-based range. EPA’s multiple RfCs for trichloroethylene (TCE) were evaluated as a case study. For TCE, a multi-endpoint safety range was judged to be 3 μg/m3 to 30 μg/m,3 based on a review of kidney effects found in NTP (1988), thymus effects found in Keil et al. (2009) and cardiac effects found in the Johnson et al. (2003) study. This multi-endpoint safety range is derived from studies for which the appropriate averaging time corresponds to different exposure durations, and, therefore, can be applied to both long- and short-term exposures with appropriate consideration of exposure averaging times. For shorter-term exposures, averaging time should be based on the time of cardiac development in humans during fetal growth, an average of approximately 20–25 days.

Aneuploidy, an imbalanced chromosome number, is associated with both cancer and developmental disorders such as Down syndrome (DS). To determine how aneuploidy affects cellular and organismal physiology, we have developed a system to evaluate aneuploid cell fitness in vivo. By transplanting hematopoietic stem cells (HSCs) into recipient mice after ablation of recipient hematopoiesis by lethal irradiation, we can directly compare the fitness of HSCs derived from a range of aneuploid mouse models with that of euploid HSCs. This experimental system can also be adapted to assess the interplay between aneuploidy and tumorigenesis. We hope that further characterization of aneuploid cells in vivo will provide insight both into the origins of hematopoietic phenotypes observed in DS individuals as well as the role of different types of aneuploid cells in the genesis of cancers of the blood., National Institutes of Health (U.S.) (Grant GM056800)

Aneuploidy, an imbalanced karyotype, is a widely observed feature of cancer cells that has long been hypothesized to promote tumorigenesis. Here we evaluate the fitness of cells with constitutional trisomy or chromosomal instability (CIN) in vivo using hematopoietic reconstitution experiments. We did not observe cancer but instead found that aneuploid hematopoietic stem cells (HSCs) exhibit decreased fitness. This reduced fitness is due at least in part to the decreased proliferative potential of aneuploid hematopoietic cells. Analyses of mice with CIN caused by a hypomorphic mutation in the gene Bub1b further support the finding that aneuploidy impairs cell proliferation in vivo. Whereas nonregenerating adult tissues are highly aneuploid in these mice, HSCs and other regenerative adult tissues are largely euploid. These findings indicate that, in vivo, mechanisms exist to select against aneuploid cells., National Institutes of Health (U.S.) (CA206157), Kathy and Curt Marble Cancer Research Fund, David H. Koch Institute for Integrative Cancer Research at MIT (Support Grant P30-CA14051), National Institute of General Medical Sciences (U.S.) (Training Grant T32GM007753)

Aneuploidy is frequently associated with disease and developmental abnormalities. It is also a key characteristic of cancer. Several model systems have been developed to study the role of chromosomal instability and aneuploidy in tumorigenesis. The results are surprisingly complex, with the conditions sometimes promoting and sometimes inhibiting tumour formation. Here, we review the effects of aneuploidy and chromosomal instability in cells and model systems of cancer, propose a model that could explain these complex findings and discuss how the aneuploid condition could be exploited in cancer therapy.

Aneuploidy decreases cellular fitness, yet it is also associated with cancer, a disease of enhanced proliferative capacity. To investigate one mechanism by which aneuploidy could contribute to tumorigenesis, we examined the effects of aneuploidy on genomic stability. We analyzed 13 budding yeast strains that carry extra copies of single chromosomes and found that all aneuploid strains exhibited one or more forms of genomic instability. Most strains displayed increased chromosome loss and mitotic recombination, as well as defective DNA damage repair. Aneuploid fission yeast strains also exhibited defects in mitotic recombination. Aneuploidy-induced genomic instability could facilitate the development of genetic alterations that drive malignant growth in cancer.

Sintering of metal nanowire (NW) networks on transparent polymers is an emerging approach for fabricating transparent conductive electrodes used in multiple devices. Pulsed light sintering is a scalable sintering process in which large-area, broad-spectrum xenon lamp light causes rapid NW fusion to increase network conductivity, while embedding the NWs in the polymer to increase mechanical robustness. This paper develops a multiphysical approach for predicting evolution of conductivity, NW fusion and nanoscale temperature gradients on the substrate during pulsed light sintering of silver NWs on polycarbonate. Model predictions are successfully validated against experimentally measured temperature and electrical resistance evolution. New insight is obtained into the diameter-dependent kinetics of NW fusion and nanoscale temperature gradients on the substrate, which are difficult to obtain experimentally. These observations also lead to the understanding that NW embedding in intense pulsed light sintering (IPL) can occur below the glass transition temperature of the polymer, and to a new differential thermal expansion-based mechanism of NW embedding during IPL. These insights, and the developed model, create a framework for physics-guided choice of NWs, substrate and process parameters to control conductivity and prevent substrate damage during the process.

Recent studies have indicated that nickel gallium alloys can be effective catalysts for the hydrogenation of CO2 to methanol. To simplify the characterization of NiGa catalysts, the authors are developing model systems using sputter deposited NiGa thin films. The NiGa thin films used in this study were deposited using an equimolar alloy target and annealed in ultrahigh vacuum. Atomic force microscopy (AFM), x-ray diffraction (XRD), and x-ray photoelectron spectroscopy (XPS) were used to characterize the NiGa films before and after annealing. AFM results showed that film roughness and grain size significantly increased as the film was annealed above 700 °C. XRD patterns indicated that NiGa thin films were nanocrystalline as deposited and then transitioned to the Ni13Ga9 phase after annealing above 500 °C. XPS results indicated that gallium and oxygen segregated to the surface after annealing up to 600 °C, and formed a surface Ga2O3 layer. For anneals above 600 °C, the Ga2O3 XPS signal was reduced in intens...

Introduction. True aneurysms of the extracranial internal carotid artery are rare lesions. Surgical treatment is considered to be the best therapeutic option. However, the use of the intraluminal shunt remains controversial. Case report. We reported a case of a giant extracranial internal carotid artery aneurysm treated by reconstructive surgery. A 76-year-old woman was referred with a pulsatile mass inside her mouth, associated with dizziness and dysarthria. There was no history of cerebrovascular symptoms, neck pain, or cervical trauma. A magnetic resonance scan showed a 45 mm aneurysm of the internal carotid artery (ICA), and kinking of ICA. Angiography demonstrated a saccular ICA aneurysm, with a lengthening and tortuosity of the ICA. The aneurysm and the carotid artery branches were easily exposed through a standard anterior cervical incision. After resection of the aneurysm, a Javid shunt was inserted between the common and internal carotid arteries, and end-to-end repair of ICA was easily performed due to ICA redundancy. The aneurysm was of atherosclerotic origin. Four months after the operation, the patient showed a complete recovery from peripheral neurological deficit. Discussion. Our results show that surgical reconstruction is a satisfactory therapeutic choice in the management of extracranial carotid artery aneurysms in order to avoid rupture, thromboembolism and cerebrovascular insufficiency. To date, there has been little experience with endoluminal exclusion techniques and the long-term effectiveness is still uncertain. .

Whole-genome sequencing, particularly in fungi, has progressed at a tremendous rate. More difficult, however, is experimental testing of the inferences about gene function that can be drawn from comparative sequence analysis alone. We present a genome-wide functional characterization of a sequenced but experimentally understudied budding yeast, Saccharomyces bayanus var. uvarum (henceforth referred to as S. bayanus), allowing us to map changes over the 20 million years that separate this organism from S. cerevisiae. We first created a suite of genetic tools to facilitate work in S. bayanus. Next, we measured the gene-expression response of S. bayanus to a diverse set of perturbations optimized using a computational approach to cover a diverse array of functionally relevant biological responses. The resulting data set reveals that gene-expression patterns are largely conserved, but significant changes may exist in regulatory networks such as carbohydrate utilization and meiosis. In addition to regulatory changes, our approach identified gene functions that have diverged. The functions of genes in core pathways are highly conserved, but we observed many changes in which genes are involved in osmotic stress, peroxisome biogenesis, and autophagy. A surprising number of genes specific to S. bayanus respond to oxidative stress, suggesting the organism may have evolved under different selection pressures than S. cerevisiae. This work expands the scope of genome-scale evolutionary studies from sequence-based analysis to rapid experimental characterization and could be adopted for functional mapping in any lineage of interest. Furthermore, our detailed characterization of S. bayanus provides a valuable resource for comparative functional genomics studies in yeast.

Polymer coatings, both with and without pigments, have been subjected to solar radiation and water spray weathering. The degrees of penetration of the weathering effects have been measured by injecting positrons of varying energy, i.e. to variable depths, into the films and observing the Doppler broadening of the annihilation radiation. The method is capable of detecting changes due to weathering effects at very early stages, long before visual examination reveals degradation. As little as one week of exposure caused measurable changes in the polymer structure, which were reflected in the Doppler broadening. Given further development, positron spectroscopy could possibly become a useful complement to the other methods of determining weatherabilities of protective polymer coatings.

Development of pathology varies widely between different strains of mice after intracerebral inoculation with the so-called 'docile' isolate of Lymphocytic Choriomeningitis (LCM) virus. The C3HeB/FeJ and B10. Br/SgSnJ mouse strains have been of special interest because they display autoimmune haemolytic anaemia with varying degrees of apparent immunological involvement. In this report, we examined the role of CD4+ T helper cells in this autoimmune response by treating mice with the CD4-specific GK1.5 monoclonal antibody. We also determined if polyclonal activation of B lymphocytes, induced either by LCM virus or by lactate dehydrogenase-elevating virus, another well known B cell activator, correlated with the development of anaemia in these mice. Our results strengthened the central role of the immune system in the anaemia in C3H mice by showing that depletion of CD4+ cells largely, if not completely, abrogated this anti-erythrocyte autoimmune reaction. As reported by others, we found that the anaemia was more mild in B10.BR mice than in C3H mice. However, we could not confirm the difference in the degree of B lymphocyte polyclonal activation between these mice. Furthermore, lactate dehydrogenase-elevating virus had no apparent effect on erythrocytes, even though this virus also induced a sharp increase in plasma IgG levels.

Natural or deliberate activation of the immune system of pathogen-free mice markedly affected their response to an autoimmune-inducing stimulus. Specifically, mice immunized with rat red blood cells were found to make antibodies reactive with both rat and mouse erythrocytes. Animals housed for an extended period in a conventional environment developed an autoimmune response twice as fast as those kept in isolators. In an attempt to emulate this effect, mice kept in a sterile environment were infected with a potent polyclonal activator of B lymphocytes, lactate dehydrogenase-elevating virus, at the same time as they were inoculated with rat erythrocytes. Whereas uninfected animals developed a progressively increasing autoantibody titer, infected mice quickly attained high anti-erythrocyte autoantibody titers that remained rather constant. Contrary to circulating autoantibodies, bound anti-erythrocyte antibodies decreased with time. Virus infection enhanced all the IgG subclass responses, with the exception of IgG1, to both rat and mouse erythrocytes. None of the modifications of the autoimmune responses resulted in anemia.

GaAs substrates were pre-patterned by electron beam (e-beam) lithography and wet-chemically etched holes. The hole structures, presented here, enable growth of site-controlled InAs quantum dots with low dot density in the range of 10^8cm^-^2 by molecular beam epitaxy (MBE). In order to reduce defect related non-radiative recombination processes at the etched surface, wet-chemical etching and a modified growth of stacked quantum dot layers are developed. The total buffer layer thickness could be increased to 55nm GaAs by only one overgrowth step of a quantum dot seed layer in comparison to strain-coupled multi-stacked structure with typically 10nm buffer layer thickness between each stack.

An aneurysm has been defined as a permanent local dilatation of the diameter of an artery by at least 50% of its normal value. A splenic artery aneurysm is most frequently a visceral artery aneurysm and clinically it is usually asymptomatic but potentially life-threatening at the same time, with the incidence of its rupturing being 2-10% and then the mortality rate ranges from 20 to 36%. A 51-year-old female patient was admitted to the Department of Vascular and Transplantation Surgery in Novi Sad having been found to have a big splenic artery aneurysm during the ultrasound examination of her abdomen after cholecystectomy. The additional diagnostic procedure - computerized tomography of the abdomen with i.v contrast subtraction angiography-confirmed the splenic artery aneurysm to have the diameter of 5 cm and therefore the elective surgical treatment was indicated after the preoperative preparation and risk assessment. The aneurysm was exposed through Chevron incision, and the detailed surgical exploration was done after the omental bursa had been opened. The aneurysmectomy and the reconstruction of the splenic artery by the termino-terminal anastomosis were performed after the weakening of the wall had been verified. The biopsies of the liver and the aneurysmal sac were done during the surgery. The pathohistological finding confirmed the atherosclerotic etiology of the aneurysm. Since the postoperative course was normal, the patient was discharged on the eighth postoperative day.

microRNAs play a critically important role in a wide array of biological processes including those implicated in cancer, neuro-degenerative and metabolic disorders, and viral infection. Although we have begun to understand microRNA biogenesis and function, experimental demonstration of their functional effects and the molecular mechanisms by which they function remains a challenge. Members of the let-7/miR-98 family play a critical role in cell cycle control with respect to differentiation and tumorigenesis. In this study, we show that exogenous addition of pre-let-7 in primary human fibroblasts results in a decrease in cell number and an increased fraction of cells in the G2/M cell cycle phase. Combining microarray techniques with DNA sequence analysis to identify potential let-7 targets, we discovered 838 genes with a let-7 binding site in their 3′-untranslated region that were down-regulated upon overexpression of let-7b. Among these genes is cdc34, the ubiquitin-conjugating enzyme of the Skp1/cullin/F-box (SCF) complex. Cdc34 protein levels are strongly down-regulated by let-7 overexpression. Reporter assays demonstrated direct regulation of the cdc34 3′-untranslated region by let-7. We hypothesized that low Cdc34 levels would result in decreased SCF activity, stabilization of the SCF target Wee1, and G2/M accumulation. Consistent with this hypothesis, small interfering RNA-mediated down-regulation of Wee1 reversed the G2/M phenotype induced by let-7 overexpression. We conclude that Cdc34 is a functional target of let-7 and that let-7 induces down-regulation of Cdc34, stabilization of the Wee1 kinase, and an increased fraction of cells in G2/M in primary fibroblasts.

Lymphocytic Choriomeningitis (LCM) virus, substrain Docile, causes a chronic infection in adult C3HeB/FeJ mice. The virus also induces a severe anemia which, unlike the viremia, eventually resolves. Initially, there is frank bone marrow deficit, but the anemia persists well beyond a strong erythroid compensatory response. An immune-mediated basis for the hemolytic anemia was suggested by its abrogation in cyclophosphamide-treated mice, as well as an abnormal number of spherocytes in the circulation. We now show by ELISA assay, using either anti-mouse Ig or RBC membrane ghosts as catching antigen, that unusually high quantities of antibodies can be eluted from the RBCs of virus-infected mice. Furthermore, the high transient antibody concentration correlates with the severity of the anemia. With no evidence for complement playing a role in the anemia, these data indicate that erythrophagocytosis (via macrophage FcRs) may be the mechanism for RBC elimination. The possibility of molecular mimicry (antibody cross-reactivity between LCM and RBC membrane epitopes) was considered but appeared unlikely since the RBC antibody eluates gave no signal in an LCM-specific ELISA (which showed an ever increasing serum titer of virus-specific antibody). Isotype determination of the RBC eluates revealed the following: IgG2a much greater than IgG1 greater than IgG2b greater than IgG3 greater than IgM. The precise role, if any, of LCM-virus induced polyclonal activation (most strikingly in the IgG2a subclass) has yet to be determined.

In the marine red alga Polyneura hilliae photoinhibition of photosynthesis was investigated by measuring the photosynthetic oxygen production. The extent of photoinhibition in this alga depends on the fluence rate as well as on the time of exposure. Strongly photoinhibited algae recover slowly. Full recovery is reached only in weak light after 3 days. Two phases of recovery can be distinguished: a first relatively fast phase of recovery that is independent of light, and a second slow phase which begins after about 6 hours and requires weak light. Consequently, even in complete darkness partial recovery is observed. An action spectrum of photoinhibition and its comparison with the in vivo absorption spectrum of Polyneura demonstrates that photoinhibition is mainly caused by light which is absorbed by phycobiliproteins, chlorophyll a and carotenoids. This fact and the ineffectiveness of light above 686 nm clearly indicate that the main photoinhibition site in this alga is PS II. No photosynthetic oxygen production is detectable after 3 h irradiation at 402 nm, 150 μmol quanta m−2s−1. As photosynthetic activity recovers slowly and to a limited extent, the suppression of oxygen production is apparently only in part the result of photoinhibition sensu strictu and in part due to permanent photodamage.

The Docile strain of lymphocytic choriomeningitis virus (LCMV) induces anemia in a number of inbred strains of mice, including C3HeB/FeJ and CBA/Ht animals. A difference in the kinetics of anemia and in compensatory reticulocytosis suggested that impaired erythropoiesis was the major pathogenic mechanism Involved in CBA/Ht mice, but not in C3HeB/FeJ mice. In both mouse strains an antierythrocyte autoantibody production that depended on the presence of functional CD4+ T lymphocytes was observed. Although depletion of T helper lymphocytes prevented anemia in C3HeB/FeJ mice, this treatment largely failed to inhibit the development of the disease in CBA/Ht animals. This observation indicated that the antierythrocyte autoimmune response induced by the infection was at least Partly responsible for the anemia of C3HeB/FeJ mice, but not of CBA/Ht mice. Erythrophagocytosis was enhanced in both mouse strains after LCMV infection, but did not appear to be a major cause of anemia. These data clearly indicate that similar disease profiles induced by the same virus in two different host strains can be the result of distinctly different mechanisms.

A monolithically integrated 5/sup th/ order lowpass filter with a tunable cutoff frequency is presented. The cutoff frequency can be tuned via a control voltage between 30 kHz and 180 kHz. This circuit may serve as channel selecting filters in direct conversion receivers for various cellular radio services with different bandwidth (e.g. IS-54, GSM, etc.). The main design goals are low power consumption, a large tuning range, good linearity and a high dynamic range.

Aortoenteric fistula represents a communication between aortic and duodenal lumen. It is a very rare complication of aortic aneurysmal disease as well as aortic prosthetic surgery. There are only few surgeons who have seen more than one case of aortoenteric fistula. At the Institute of Surgery one case of aortoenteric fistula was treated. It was well diagnosed at the Institute for Internal Medicine, and then presented to the vascular surgeon who operated immediately. The whole diagnostic and therapeutic procedure was well done, but stenosing carcinoma of ascendent colon was found during the operation. Resection were done leading to the dehiscence of bowel anastomosis and fatal outcome for the patient. This case is instructive in a diagnostic and surgical manner.

The effect of LCMV on CD4+ T lymphocytes was analyzed in C3HeB/FeJ mice after infection with the Docile strain of this virus. Our results indicated that LCMV triggers: i) an inhibition of Th2 lymphocyte differentiation induced by concomitant immunization with a nonviral protein antigen; ii) a depression of T helper-dependent antibody responses elicited by such an immunization; and iii) a CD4+ cell-mediated proliferation of spleen cells leading to increased interleukin-4 and interferon-gamma message expression and IgG2a-restricted total immunoglobulin secretion. Taken together, these results indicate that LCMV profoundly affects CD4+ cell-mediated immune responses in infected animals. Such modulations of T-helper functions may explain the preponderance of IgG2a in the antierythrocyte autoimmune response induced by the virus in C3HeB/FeJ mice.

Background. Healthy Start is a 3-year demonstration and education research project designed to evaluate the effectiveness of a multidimensional cardiovascular (CV) risk reduction intervention in preschool centers over a 3-year period of time. Methods. Two primary interventions are employed. The first is the preschool food service intervention program designed to reduce the total fat in preschool meals and snacks to less than 30% of calories and reduce the saturated fat to less than 10% of calories. The second major intervention is a comprehensive preschool health education curriculum, focused heavily on nutrition. Results. Effectiveness of the intervention will be determined through evaluation of changes in dietary intake of preschool children at school meals and snacks, especially with respect to intake of total and saturated fat. Evaluation of the education component will include assessment of program implementation by teachers, assessment of changes in nutrition knowledge by preschool children, and assessment of changes in home meals that children consume (total and saturated fat content). Blood cholesterol will be evaluated semiannually to evaluate changes that may be due to modification of dietary intake. Growth and body fatness will also be assessed. Conclusions. While substantial efforts have targeted CV risk reduction and health education for elementary school children, similar efforts aimed at preschool children have been lacking. The rationale for beginning CV risk reduction programs for preschool children is based upon the premise that risk factors for heart disease are prevalent by 3 years of age and tend to track over time, most commonly hypercholesterolemia and obesity, both related to nutrition. Since the behavioral antecedents for nutritional risk factors begin to be established very early in life, it is important to develop and evaluate new educational initiatives such as Healthy Start , aimed at the primary prevention of cardiovascular risk factors in preschool children. The purpose of this publication is to describe the rationale and methods for the Healthy Start project.

This paper analyzes results of 5-year surgical treatment of patients with ruptured abdominal aorta aneurysms (1991.-1995.) at the Clinic for Vascular and Transplantation Surgery of the Institute of Surgery in Novi Sad. 105 patients with abdominal aorta aneurysm underwent surgery, whereas in 31 patients there was a suspicion of rupture and it was confirmed by US and CT examination. One of basic factors to decrease mortality in these patients is early diagnosis and surgery before hemorrhagic shock occurs. Results in hemodynamic stabile patients with blood pressure over 100 mmHg and regular diuresis are much better with mortality of 20%. In order to estimate the correlation of hemodynamic state and outcome of the operation, patients were divided into three groups--hemodynamic stable with blood pressure over 100 mmHg and regular diuresis at admission: hemodynamic unstable patients with signs of mild or moderate shock and blood pressure under 100 mmHg and without initial diuresis which was regulated at the beginning of therapy and hemodynamic unstable patients in severe shock and unmeasurable blood pressure. The highest survival rate (10% mortality) and the least complications occurred in the first group of patients. The total mortality of patients after surgery was 48.48%. Timely diagnosis, suspicion of rupture and adequate first and with urgent transfer to a competent surgical institution are key factors in treatment of this disease and its outcome.

C3HeB/FeJ mice infected with the docile strain of lymphocytic choriomeningitis virus (LCMV-d) develop a persistent infection with a transient haemolytic anaemia. Immunoglobulin can be eluted from the red blood cells (RBC) of these mice but it cannot be detected on the RBC by a conventional antiglobulin test. The present study demonstrates that RBC from such mice bear erythrocyte autoantibodies which are predominantly of the IgG2a subclass, with lower levels of autoantibodies of the IgG1, IgG2b and IgG3 subclasses. To identify the target antigen the autoantibodies were eluted from the RBC of LCMV-infected mice. The eluted autoantibody bound to intact normal RBC and precipitated a 105000 MW component that corresponds to murine Band 3 protein. A monoclonal antibody derived from mice infected with LCMV-d also precipitated mouse Band 3, and reacted specifically by enzyme-linked immunosorbent assay against a purified preparation of Band 3. This study has shown that in C3H mice infected with LCMV-d which develop autoimmune haemolytic anaemia, the target autoantigen is erythrocyte membrane Band 3.