Contrast-enhanced computed tomography as a preoperative indicator of vascular invasion from adrenal masses in dogs.
The purpose of this study was to determine if contrast-enhanced computed tomography (CT) accurately reflected vascular luminal invasion by adrenal masses in dogs. Medical records of 15 dogs with 17 adrenal masses that underwent preoperative CT imaging were reviewed. Presence or absence of vascular invasion was confirmed by surgery or necropsy/histology. Vascular invasion was identified correctly using contrast-enhanced CT in 11/12 dogs. The sensitivity and specificity of contrast-enhanced CT for vascular invasion compared with surgery or necropsy was 92% and 100%, respectively. The positive predictive value and negative predictive value of contrast-enhanced CT was 100% and 90%, respectively. The accuracy was 95%. In one dog, invasion of the phrenicoabdominal vein was not identified on CT images. Six of eight masses with vascular invasion where a histologic diagnosis was obtained were malignant. Four of four pheochromocytomas invaded adjacent vasculature. Contrast-enhanced CT provided accurate preoperative assessment of adrenal masses. Vascular invasion by adrenal masses in this study occurred by way of the lumen of the phrenicoabdominal vein with subsequent intraluminal extension into other veins, rather than by erosion through vessel walls.


Growth hormone-dependent and -independent regulation of cytochrome P-450 isozyme expression in streptozotocin-diabetic rats.
The sexually dimorphic GH secretory pattern is thought to be the major factor regulating constitutive expression of hepatic P450IIC11 (P-450h) and P450IIC12 (P-450i). In this study we investigated whether factors other than the diabetes-induced decrease in GH secretion contribute to alterations in P-450 isozyme expression in streptozotocin (STZ)-diabetic rats. In male rats, hepatic P-450h apoprotein and mRNA decreased to 13% and 24% of control male levels, respectively, within 14 days of STZ injection. STZ-diabetes had little effect on expression of P-450i in females. Treatment of diabetic male rats with GH did not reverse the suppression of P-450h. STZ treatment also suppressed P-450h expression in GH-treated hypophysectomized (Hx) male rats, but incompletely. Thus, GH can partially reverse diabetic suppression of P-450h. However, in Hx male rats without GH supplementation, STZ treatment suppressed P-450h apoprotein and mRNA expression to 16% and 6% of nondiabetic Hx male levels, respectively, demonstrating the existence of GH-independent regulation of P-450h expression. In Hx female rats, P-450h apoprotein levels were 40% of those in intact control males and were not significantly decreased by STZ. Concomitantly, STZ produced a greater decrease in serum insulin levels and a greater increase in serum glucagon in Hx male rats than in Hx females. The results provide evidence for the existence of STZ-sensitive GH-independent expression of P-450h and further document the gender differences in STZ sensitivity.


Parental imprinting of the mouse H19 gene.
THE mouse H19 gene encodes one of the most abundant RNAs in the developing mouse embryo. It is expressed at the blastocyst stage of development, and accumulates to high levels in tissues of endodermal and mesodermal origin (H. Kim, unpublished result). After birth the gene is expressed in all tissues except skeletal muscle. It lacks a common open reading frame in the 2.5-kilobase RNA, but has considerable nucleotide sequence similarity between the genes of rodents and humans. Expression of the gene in transgenic mice results in late prenatal lethality, suggesting that the dosage of its gene product is strictly controlled. The H19 gene maps to the distal segment of mouse chromosome 7, in a region that is parentally imprinted, a process by which genes are differentially expressed on the maternal and paternal chromosomes. We have now used an RNase protection assay that can distinguish between H19 alleles in four subspecies of Mus, to demonstrate that the H19 gene is parentally imprinted, with the active copy derived from the mother. This assay will be of general use in assaying allele-specific gene expression.


Daytime radiative cooler using porous TiO2: new approach.
Passive daytime radiative cooling remains a topic of intense interest that has gained great attention recently, mainly due to its important applications, such as electronic heat dissipation, solar cells, and photo-thermal technologies. In order to achieve the daytime radiative cooling for thermal management applications, a new smart shield should be designed to have both a low absorptivity in the solar range of 0.3-2.5 µm and high emissivity in the atmospheric transparency windows of 8-13 µm and 16-26 µm. In this work, we propose and analyze a new, inexpensive, and highly scalable double-layer coating to reach an efficient passive daytime radiative cooling. Double-layer coatings consisting of a top TiO2 porous adsorbent layer and a bottom providing excellent transmittance in the solar spectrum are achieved properly by borosilicate-crown glass (BK7). We have shown that the proposed device preserves an average absorptivity/emissivity well below 7% in the entire solar spectrum and almost 95% in the atmospheric transparency windows. Besides, our design allows for sub-ambient temperature drops of 45° and diurnal cooling power output of 133W/m2, even in the case of solar irradiance up to 1000W/m2. The cooling performance persists under high humidity, even when taking the non-radiative heat exchange progress into account. The results revealed that the proposed design can be easily applied to a large area and promotes a significant step towards achieving large scale application in solar cells and related systems.


An Accurate Model for the Ion Current-Distance Behavior in Scanning Ion Conductance Microscopy Allows for Calibration of Pipet Tip Geometry and Tip-Sample Distance.
The scanning ion conductance microscope (SICM) is an emerging tool for noncontact topography imaging and multiphysical investigation of soft samples in aqueous environments such as living cells. Despite the increasing popularity of SICM, several aspects of the imaging process are still unknown; for example, there is still no accurate description of the behavior of the ion current for a varying tip-sample distance. To predict this ion current-distance behavior, we provide a new numerical model based on finite element modeling. The model allows, for the first time, accurately determining the tip-sample distance during an SICM experiment. Furthermore, we present a nondestructive method for calibrating the pipet tip geometry by fitting the numerical model to the experimental ion current-distance data and verify this method using pipets with opening radii between 30 and 300 nm.


Loss of pluripotency in human embryonic stem cells directly correlates with an increase in nuclear zinc.
The pluripotency of human embryonic stem cells (hESCs) is important to investigations of early development and to cell replacement therapy, but the mechanism behind pluripotency is incompletely understood. Zinc has been shown to play a key role in differentiation of non-pluripotent cell types, but here its role in hESCs is directly examined. By mapping the distribution of metals in hESCs at high resolution by x-ray fluorescence microprobe (XFM) and by analyzing subcellular metal content, we have found evidence that loss of pluripotency is directly correlated with an increase in nuclear zinc. Zinc elevation not only redefines our understanding of the mechanisms that support pluripotency, but also may act as a biomarker and an intervention point for stem cell differentiation.


MEMRI and tumors: a method for the evaluation of the contribution of Mn(II) ions in the extracellular compartment.
The purpose of the work was to set-up a simple method to evaluate the contribution of Mn(2+) ions in the intra- and extracellular tumor compartments in a MEMRI experiment. This task has been tackled by "silencing" the relaxation enhancement arising from Mn(2+) ions in the extracellular space. In vitro relaxometric measurements allowed assessment of the sequestering activity of DO2A (1,4,7,10-tetraazacyclododecane-1,7-diacetic acid) towards Mn(2+) ions, as the addition of Ca-DO2A to a solution of MnCl2 causes a drop of relaxivity upon the formation of the highly stable and low-relaxivity Mn-DO2A. It has been proved that the sequestering ability of DO2A towards Mn(2+) ions is also fully effective in the presence of serum albumin. Moreover, it has been shown that Mn-DO2A does not enter cell membranes, nor does the presence of Ca-DO2A in the extracellular space prompt migration of Mn ions from the intracellular compartment. On this basis the in vivo, instantaneous, drop in SE% (percent signal enhancement) in T1 -weighted images is taken as evidence of the sequestration of extracellular Mn(2+) ions upon addition of Ca-DO2A. By applying the method to B16F10 tumor bearing mice, T1 decrease is readily detected in the tumor region, whereas a negligible change in SE% is observed in kidneys, liver and muscle. The relaxometric MRI results have been validated by ICP-MS measurements.


Doxorubicin and octreotide induce a 40 kDa breakdown product of p53 in human hepatoma and tumoral colon cell lines.
The chemotherapeutic drug doxorubicin and the anti-proliferative long-acting somatostatin analogue octreotide, both used in cancer treatment, have been shown to increase the expression of the p53 tumour suppressor protein. In the present study, we demonstrate by Western-blot analysis that, in addition to the p53 protein, these molecules were able to induce the expression of a shorter protein with an apparent molecular mass of 40 kDa (p40), recognized by antibodies raised against the N-terminus of p53. This induction was present in tumoral and non-tumoral cells and did not depend on the status of the endogenous p53 protein. The p40 protein was significantly induced after 3 h of cell treatment with doxorubicin or octreotide, remained stable until 24 h and was located in the nuclear extract. Using reverse primers corresponding to each exon of the p53 gene, only one transcript was amplified by reverse transcriptase-PCR. This suggested that p40 was issued from a post-translational modification and not from an alternative splicing. This protein was not recognized by the PAb421 antibody, suggesting that it was issued from a cleavage of the p53 C-terminal region (p40deltaC). Furthermore, this cleavage was not dependent on caspase activity. In conclusion, these results support the hypothesis that this post-translational modification plays a significant role in the regulation of multiple p53 signalling pathways. These results also suggest that octreotide, a molecule with different signalling pathways, was able as doxorubicin to generate a p53 breakdown product.


Acute stress-induced increases in exhaled nitric oxide in asthma and their association with endogenous cortisol.
psychosocial stress is known to influence the pathophysiology of asthma. Although stress has been linked to serum markers of inflammatory activity and exaggerated response to allergen challenge in asthma, few studies have examined inflammatory activity in the airways linked to psychosocial stress alone. Furthermore, although studies have demonstrated lower levels or reactivity of endogenous cortisol in asthma, the association with airway inflammatory activity in stress remains unexplored. we therefore studied airway inflammation and cortisol response to a standardized laboratory task inducing acute psychosocial stress. airway inflammation by the fraction of exhaled nitric oxide (FeNO) and saliva cortisol were sampled before and up to 45 minutes after experimental challenge with the Trier Social Stress Test in 20 adult patients with asthma and 19 healthy control subjects. Respiratory inductive plethysmography was used to control for changes in ventilatory activity that impact FeNO levels. FeNO levels were generally higher in patients with asthma than healthy control subjects and salivary cortisol levels were lower. Increases in cortisol levels were observed after the stress protocol in both groups (P < 0.001). FeNO levels at the time of peak cortisol increase after stress were significantly higher than before stress in both groups (P < 0.05). FeNO increases were independent of changes in ventilation. In patients with asthma, higher cortisol levels and stronger increases in cortisol after stress were significantly associated with smaller increases in FeNO (P < 0.05). acute psychosocial stress alone increases airway inflammatory markers and this increase is attenuated by stronger stress-related activity of the hypothalamic-pituitary-adrenal axis in asthma.


Persistent and transient replication of full-length hepatitis C virus genomes in cell culture.
The recently developed subgenomic hepatitis C virus (HCV) replicons were limited by the fact that the sequence encoding the structural proteins was missing. Therefore, important information about a possible influence of these proteins on replication and pathogenesis and about the mechanism of virus formation could not be obtained. Taking advantage of three cell culture-adaptive mutations that enhance RNA replication synergistically, we generated selectable full-length HCV genomes that amplify to high levels in the human hepatoma cell line Huh-7 and can be stably propagated for more than 6 months. The structural proteins are efficiently expressed, with the viral glycoproteins E1 and E2 forming heterodimers which are stable under nondenaturing conditions. No disulfide-linked glycoprotein aggregates were observed, suggesting that the envelope proteins fold productively. Electron microscopy studies indicate that cell lines harboring these full-length HCV RNAs contain lipid droplets. The majority of the core protein was found on the surfaces of these structures, whereas the glycoproteins appear to localize to the endoplasmic reticulum and cis-Golgi compartments. In agreement with this distribution, no endoglycosidase H-resistant forms of these proteins were detectable. In a search for the production of viral particles, we noticed that these cells release substantial amounts of nuclease-resistant HCV RNA-containing structures with a buoyant density of 1.04 to 1.1 g/ml in iodixanol gradients. The same observation was made in transient-replication assays using an authentic highly adapted full-length HCV genome that lacks heterologous sequences. However, the fact that comparable amounts of such RNA-containing structures were found in the supernatant of cells carrying subgenomic replicons demonstrates a nonspecific release independent of the presence of the structural proteins. These results suggest that Huh-7 cells lack host cell factors that are important for virus particle assembly and/or release.


Improvement of Growth, Yield, Seed Production and Phytochemical Properties of Satureja khuzistanica Jamzad by Foliar Application of Boron and Zinc.
Satureja khuzistanica Jamzad is a valuable and endemic medicinal plant. Boron and zinc are essential elements for the vegetative and reproductive growth of plants and have significant effects on yield, essential oil composition and the seed production of plants. To investigate the effects of the foliar application of zinc and boron on the growth, yield, seed production and phytochemical properties of S. khuzistanica, a study was conducted in a factorial experiment with three replicates in two consecutive years based on a randomized complete block design. The foliar application of boron (B) at three concentrations (control or distilled water, 0.4% and 0.8% as H3BO3) and zinc (Zn) at three concentrations (control or distilled water, 0.3% and 0.6% as ZnSO4) was carried out. Our results showed that the foliar application of B resulted in a significant increase in the fresh and dry weights of plants, the dry weight of stems, drug yield, seed yield, seed germination and 1000-seed weight. At the same time, the application of B resulted in a significant decrease in seed emptiness. The fresh and dry weights of plants, drug yield, seed yield, 1000-seed weight and seed germination were also significantly improved by Zn foliar spraying compared to the control. Application of 0.8% B resulted in a significant decrease in seed emptiness by 14.16% and 22.37%, as compared to the control. The foliar spraying of B and Zn improved the total phenolic content, the essential oil content and the yield and antioxidant activity of S. khuzistanica. Moreover, B application generally concentrated more carvacrol in the essential oil (in the first experimental year). In contrast, no significant differences were observed between Zn treatments in carvacrol content and total flavonoids. The use of several microelements, such as B and Zn, could improve both the quantity and quality of S. khuzistanica. Additionally, improvement of seed set and seed quality by the foliar spraying of Zn and B may be useful for growing plants in arid and semi-arid areas.


Care home medicine.
Care homes are changing, with improved standards and expanding roles, and it is vital that their residents are not excluded or discriminated against by health and social services. Staff are keen to be educated, and investment of time and money is rewarded with improved outcomes in many areas.


Staging of urinary bladder neoplasms with MR imaging: is Gd-DTPA helpful?
This study was performed to investigate whether intravenous administration of Gd-DTPA can improve the accuracy of MR imaging in the detection and staging of bladder neoplasms. In 68 patients with suspected urinary bladder neoplasms, MR examinations were performed with T1-weighted SE sequences before and after intravenous administration of Gd-DTPA. The findings were compared with surgical staging using the TNM classification. Overall staging accuracy of contrast enhanced MR was 46%; if stages Ta-T3a were combined into one group, the accuracy was 69%. Accuracy was low (19%) in tumors without muscular bladder wall invasion (Ta). In cases with extravesical spread (greater than or equal to T3b), the accuracy of staging was 87%. Contrast enhanced MR detected extravesical extension of tumor with a sensitivity of 93% and a specificity of 95%. Contrast enhancement increased the sensitivity for detection of urinary bladder neoplasms from 70% on precontrast T1-weighted scans to 79% on postcontrast scans. In comparison with T2-weighted scans, the Gd-DTPA enhanced T1-weighted scans had better image quality and lower acquisition times.


Tracheal Intubation Using Nelaton Catheter: A Simple Device, an Airway Rescue Option.
Although anticipated, difficult airway could turn into a catastrophe for health care providers, where planned strategies and equipment become useless. We report a challenging airway management process due to narrowed upper airway by the application of a Nelaton catheter as a rescue device.


Transcriptional regulation of ULBP1, a human ligand of the NKG2D receptor.
Tumor cells expressing ligands of the NKG2D receptor stimulate anti-tumor immunity mediated by natural killer and T cells. In humans, NKG2D ligands (NKG2DL) are encoded by MIC and ULBP proteins. NKG2DL exhibit highly restricted expression in healthy tissues but are widely expressed in tumors. However, regulation of each NKG2DL differs substantially in different cancer cells. In this study, we characterized the mechanisms that regulate the expression of ULBP1. We show that the transcription of ULBP1 strictly depends on the binding of Sp1 and Sp3 to a CRE(1) site located in the ULBP1 minimal promoter. The mutation or deletion of this Sp1/Sp3 binding site abolished the transcription of ULBP1. It also diminished the transactivation of ULBP1 promoter by Sp3 overexpression, but not by Sp1, indicating that Sp3 is the main transcription factor that regulates ULBP1 through the CRE(1) site. Experiments in SL2 cells showed that the ULBP1 promoter was inactive in the absence of the Sp proteins and indicate that Sp3 is the essential activator of ULBP1 transcription, because the overexpression of Sp3 up-regulated its promoter activity > 500-fold. Additionally, we demonstrated that AP-2alpha repressed the expression of ULBP1 in HeLa cells by interfering with the binding of Sp3 and Sp1 to the ULBP1 promoter. These data indicate that Sp1, Sp3, and AP-2alpha may play an important role in the immunosurveillance against cancer. Finally, the definition of ULBP1 regulation may have implications for development of new therapeutic strategies against cancer cells.


Decomposing the variability of match physical performance in professional soccer: Implications for monitoring individuals.
The aims of this study were to establish sources of variability in match physical performance of professional soccer players and provide a method for monitoring individual between-match changes. Eleven players meeting the final inclusion criteria were monitored through an entire in-season competition phase (n = 240 individual match observations). Ten Hertz global positioning systems were used to measure match total distance (TD), total high-speed running distance (≥ 21 km·h-1; HSRD), total accelerations (TAcc) and maximum running velocity (Vmax). Between-player, between-position, between-match and within-player variability were determined through linear mixed effects models. These data were then used to establish the practical significance of individual changes using a Minimum Effects Testing framework. All sources of variability were greater for HSRD (13-36%) when compared with all other metrics (<6%). Using combined between-match and within-player variability along with the smallest worthwhile change (0.2 × between-player SD), between-match individual changes of ±∼10-15% in TD, TAcc and Vmax were established as practically significant. For HSRD, these thresholds were considerably higher (≥60%). In conclusion, the ability for soccer practitioners to identify meaningful changes in match physical performance can aid decision making around player management following competition. Our study provides a method to flag changes beyond the normal match-to-match variability and by a substantial magnitude. This may have implications for recovery but should be combined with other sources of data (internal load and response) and used only as an adjunct to practitioner domain knowledge/experience.


Centromeres: the wild west of the post-genomic age.
Centromeric repetitive DNA is the largest missing piece of the Human Genome Project. Rather than being necessary or sufficient to specify the site of mitotic spindle attachment to the chromosome, centromeric DNA sequence is all but irrelevant. Instead, centromeres are thought to be specified by a protein component, a histone H3 variant called Centromere Protein A (CENP-A). This review includes a brief overview of the history of the centromere field, and the current status of knowledge on CENP-A assembly. New evidence for CENP-A recruitment in response to DNA damage implies a mechanism for neocentromere formation, and raises new questions about the epigenetic model of centromere maintenance.


Genetic diversity of the potentially therapeutic tapeworm Hymenolepis diminuta (Cestoda: Cyclophyllidea).
The cestode Hymenolepis diminuta is highly prevalent in wild rat populations and has also been observed rarely in humans, generally causing no apparent harm. The organism has been studied for decades in the laboratory, and its colonization of laboratory rats has recently been shown as protective against some inflammation-associated disorders. Recently, H. diminuta has become a leading candidate for helminth therapy, an emerging method of "biota enrichment" used to treat or prevent inflammatory diseases of humans in Western society. While most of the experimental isolates of H. diminuta are identified based on typical morphological features, hymenolepidid tapeworms may represent complexes of cryptic species as detected by molecular sequence data. In the present study, we explored the diversity of laboratory-kept strains using partial sequences of two genes (lsrDNA and cox1) and determined that H. diminuta isolates currently considered for therapeutic purposes in the US and Europe belong to a single, genetically nearly uniform lineage, showing only little genetic deviation from wild-caught isolates.


[A case of mitochondrial cytopathy (MERFF) without ragged red fibers at the onset].
We report a case of a mitochondrial cytopathy (MERRF) with a late diagnosis because of the absence of Ragged-Red Fibers (RRF) in the first muscular biopsy performed in childhood. Eight years after the onset of the disease the familial history and the clinical manifestations were very suggestive of a mitochondrial cytopathy and a second biopsy was performed confirming the diagnosis of MERRF. The authors discuss several hypotheses to explain the negativity of the first muscular biopsy. The absence of ragged-red fibers does not exclude the diagnosis of a mitochondrial cytopathy and other investigations are justified such as the biochemical study of the respiratory chain and the molecular analysis of mitochondrial DNA.


Multiple cytoskeletal pathways and PI3K signaling mediate CDC-42-induced neuronal protrusion in C. elegans.
Rho GTPases are key regulators of cellular protrusion and are involved in many developmental events including axon guidance during nervous system development. Rho GTPase pathways display functional redundancy in developmental events, including axon guidance. Therefore, their roles can often be masked when using simple loss-of-function genetic approaches. As a complement to loss-of-function genetics, we constructed a constitutively activated CDC-42(G12V) expressed in C. elegans neurons. CDC-42(G12V) drove the formation of ectopic lamellipodial and filopodial protrusions in the PDE neurons, which resembled protrusions normally found on migrating growth cones of axons. We then used a candidate gene approach to identify molecules that mediate CDC-42(G12V)-induced ectopic protrusions by determining if loss of function of the genes could suppress CDC-42(G12V). Using this approach, we identified 3 cytoskeletal pathways previously implicated in axon guidance, the Arp2/3 complex, UNC-115/abLIM, and UNC-43/Ena. We also identified the Nck-interacting kinase MIG-15/NIK and p21-activated kinases (PAKs), also implicated in axon guidance. Finally, PI3K signaling was required, specifically the Rictor/mTORC2 branch but not the mTORC1 branch that has been implicated in other aspects of PI3K signaling including stress and aging. Our results indicate that multiple pathways can mediate CDC-42-induced neuronal protrusions that might be relevant to growth cone protrusions during axon pathfinding. Each of these pathways involves Rac GTPases, which might serve to integrate the pathways and coordinate the multiple CDC-42 pathways. These pathways might be relevant to developmental events such as axon pathfinding as well as disease states such as metastatic melanoma.


[Effects of interleukin-1 receptor antagonist on the apoptosis of eosinophil in guinea pig with asthma].
To evaluate the effect of interleukin-1 receptor antagonist(IL-1ra) on apoptosis and associated mechanism of eosinophil in guinea pig with asthma. A model of guinea pig with asthma was established. After inhalation of different concentrations of IL-1ra, asthma was induced in the guinea pig for 8 days, the concentration of eosinophil cationic protein (ECP) in serum and bronchoalveolar lavage fluid (BALF), IL-5 in serum, the eosinophil counts and apoptosis were assayed by radioimmunology, enzyme-linked immunosorbent assay(ELISA), fluoromicroscope and light microscope. IL-1ra indirectly decreased the level of IL-5 in serum, improved the apoptosis of eosinophil(EOS) in lung, then decreased the level of ECP in serum and BALF. Inhalation of nebulized IL-1ra showed protective effect against asthma through change of the activity and infiltration of EOS in lung.


Relationship between residential district and health-related quality of life in Chungnam industrial complex area.
This study aimed to evaluate the relationship between residential district of people, such as power plant, steel-mill and petrochemical industries, and health-related quality of life (HRQoL). Using a cross-sectional study design, we randomly recruited participants for our study from industrial areas (thermoelectric power plant, steel-mill, petrochemical industry) and rural areas. Logistic regression analysis was used to identify the relationships between Euro quality of life-5 dimension (EQ-5D) scores and living region, while controlling for sociodemographic characteristics. In adjusted model, quality of life decreased with increasing category of age and were lower for females than males. EQ-5D scores of people living in the vicinity of thermoelectric power plant were significant lower than those of people living the vicinity of comparison region (odds ratio, 1.59; 95% confidence interval, 1.00 to 2.53). Living region of thermoelectric power plant, was strongly associated with scores on the EQ-5D. More research is needed to elucidate the mechanisms which makes the relationship with the living regions and HRQoL.


Optimized strategies for sequence-tagged-site selection in genome mapping.
The physical mapping of complex genomes is based on the construction of a genomic library and the determination of the overlaps between the inserts of the mapping clones in order to generate an ordered, cloned representation of nearly all the sequences present in the target genome. Evaluation of the relative efficiency of experimental procedures used to accomplish this goal must minimally include a comparison of the fraction of the genome covered by the ordered arrays (or "contigs"), the average size of the contigs, and the cost, in terms of time and resources, required to generate the map. Sequence-tagged-site (STS) content mapping is one strategy that has been proposed and is being utilized for this type of experiment. This paper describes three STS selection schemes and presents computer simulations of contig-building experiments based on these procedures. The results of these simulations suggest that a nonrandom STS strategy that uses paired probes requires one-third to one-fourth as many STS assays as are required in random and nonpaired approaches, and also results in a map that has both greater genome coverage and a larger average contig size. This strategy promises to reduce the time and cost required to build a high-quality physical map.


Immunological and HPLC detection of aflatoxin adducts in human tissues after an acute poisoning incident in S.E. Asia.
Acid hydrolysed, purified DNA, extracted from formalin fixed human tissues from persons acutely exposed to aflatoxins during a poisoning incident, was found to inhibit antibody binding in a competitive aflatoxin inhibition ELISA both before and after immunoaffinity column purification. HPLC analysis of acid hydrolysates of the DNA revealed a peak with a retention time 3 min earlier than 8,9-dihydro-2-(N7-guanyl)-9-hydroxy AFB1 synthesized either by peracid activation or direct reaction of the 8,9-oxide with DNA. The major peak seen when DNA was extracted from formalin fixed tissues from rats treated with aflatoxin B1 was identical to that seen in the formalin fixed human tissues. Adduct levels ranged from 0 to 170 x 10(6) nucleotides depending on tissue type and individual examined.


Long-term outcome of ketoconazole and tacrolimus co-administration in kidney transplant patients.
To study the long-term outcome of ketoconazole and tacrolimus combination in kidney transplant recipients. From 2006 to 2010, ketoconazole was given in 199 patients and was continued for at least 1 year or until graft failure (Group 1), while 149 patients did not receive any ketoconazole (Group 2). A combination of tacrolimus, mycophenolate and steroid was used as maintenance therapy. High risk patients received basiliximab induction. Basic demographic data was similar between the 2 groups. The 5-year cumulative incidence of biopsy-confirmed and clinically-treated acute rejection was significantly higher in Group 1 than in Group 2 (34% vs 18%, P = 0.01). The 5-year Kaplan-Meier estimated graft survival (74.3% vs 76.4%, P = 0.58) and patient survival (87.8% vs 87.5%, P = 0.93) were not different between the 2 groups. Multivariable analyses identified ketoconazole usage as an independent risk of acute rejection (HR = 2.33, 95%CI: 1.33-4.07; P = 0.003) while tacrolimus dose in the 2(nd) month was protective (HR = 0.89, 95%CI: 0.75-0.96; P = 0.041). Co-administration of ketoconazole and tacrolimus is associated with significantly higher incidence of acute rejection in kidney transplant recipients.


In vivo effects of Ashwagandha (Withania somnifera) extract on the activation of lymphocytes.
This study investigated the immunologic effects of Ashwagandha (Withania somnifera) on four types of immune cells in a human sample to determine the immunologic mechanism. Five (5) participants consumed 6 mL of an Ashwagandha root extract twice daily for 96 hours. Ashwagandha was administered with anupana (whole milk). Peripheral blood samples were collected at 0, 24, and 96 hours and compared for differences in cell surface expression of CD4, CD8, CD19, CD56, and CD69 receptors by flow cytometry. Significant increases were observed in the expression of CD4 on CD3+ T cells after 96 hours. CD56+ NK cells were also activated after 96 hours as evidenced by expression of the CD69 receptor. At 96 hours of use, mean values of receptor expression for all measured receptor types were increased over baseline, indicating that a major change in immune cell activation occurred across the sample. Effects on immune cell activation with use of Ashwagandha warrant further study.


Electroconvulsive therapy for the treatment of refractory mania.
We present the case of a patient with treatment-refractory mania. The patient had been tried on numerous medications, to which she either did not respond well or on which she developed severe side effects, However, the patient improved rapidly when treated with unilateral electropercussive therapy (ECT) following a court order. We outline the legal barriers that have been raised against the use of ECT in patients with mania, who often refuse treatment, and the irony that ECT can be safer than medications for some patients. ECT is underutilized in mania but deserves more frequent consideration. (Journal of Psychiatric Practice. 2011;17:61-66).


Characterization of trichome-specific BAHD acyltransferases involved in acylsugar biosynthesis in Nicotiana tabacum.
Glandular trichomes of tobacco (Nicotiana tabacum) produce blends of acylsucroses that contribute to defence against pathogens and herbivorous insects, but the mechanism of assembly of these acylsugars has not yet been determined. In this study, we isolated and characterized two trichome-specific acylsugar acyltransferases that are localized in the endoplasmic reticulum, NtASAT1 and NtASAT2. They sequentially catalyse two additive steps of acyl donors to sucrose to produce di-acylsucrose. Knocking out of NtASAT1 or NtASAT2 resulted in deficiency of acylsucrose; however, there was no effect on acylsugar accumulation in plants overexpressing NtASAT1 or NtASAT2. Genomic analysis and profiling revealed that NtASATs originated from the T subgenome, which is derived from the acylsugar-producing diploid ancestor N. tomentosiformis. Our identification of NtASAT1 and NtASAT2 as enzymes involved in acylsugar assembly in tobacco potentially provides a new approach and target genes for improving crop resistance against pathogens and insects.


TMBP200, a XMAP215 homologue of tobacco BY-2 cells, has an essential role in plant mitosis.
TMBP200 from tobacco BY-2 cells is a member of the highly conserved family of microtubule-associated proteins that includes Xenopus XMAP215, human TOGp, and Arabidopsis MOR1/GEM1. XMAP215 homologues have an essential role in spindle assembly and function in animals and yeast, but their role in plant mitosis is not fully clarified. Here, we show by immunoblot analysis that TMBP200 levels in synchronously cultured BY-2 cells increased when the cells entered mitosis, thus indicating that TMBP200 plays an important role in mitosis in tobacco. To investigate the role of TMBP200 in mitosis, we employed inducible RNA interference to silence TMBP200 expression in BY-2 cells. The resulting depletion of TMBP200 caused severe defects in bipolar spindle formation and resulted in the appearance of multinucleated cells with variable-sized nuclei. This finding indicates that TMBP200 has an essential role in bipolar spindle formation and function.


Novel 175Yb-Poly (L-Lactic Acid) Microspheres for Transarterial Radioembolization of Unrespectable Hepatocellular Carcinoma.
Novel biodegradable Poly (L-lactic acid) (PLLA) microspheres containing ytterbium were designed for intra-tumoral radiotherapy, especially for radioembolization. 175Yb possess both therapeutic beta and diagnostic gamma radiations. In this work, a process of making ready radiomicrospheres 175Yb (acac)3-loaded PLLA for more consideration has been investigated. The radiomicrospheres were prepared with approximate size of 20-40 µm, and radionuclidic purity > 92%. The radiomicrospheres were stable in-vitro for up to 72 h in normal saline, and also in human serum albumin (HSA). Biodistribution in mice bearing 4T1 tumor showed specific radionuclide uptake over 48 h. Tumor necrosis was also observed at the injection site up to 12 days after injection. These data indicated that 175Yb-PLLA microspheres could be prepared and considered further for radiomicrospheres tumor therapy.


Kidney cachexia or protein-energy wasting in chronic kidney disease: facts and numbers.
Weight loss and homeostatic disturbances of both energy and protein balances are characteristics of several illnesses including cancer, heart failure, and chronic kidney disease (CKD). Different definitions have been used to describe this deleterious process. The term protein-energy wasting (PEW) has been proposed for CKD patients by the International Society of Renal Nutrition and Metabolism. We searched the publication in Medline from February 2008 to September 2018 using PEW or cachexia in their title. Since its inception, the term PEW has been exceptionally successful, highlighted by 327 original publications referenced in PubMed over 10 years. Using this classification, several studies have confirmed that PEW is among the strongest predictors of mortality in CKD patients [hazard ratio of 3.03; confidence interval of 1.69-5.26 in 1068 haemodialysis patients and 1.40 (1.04-1.89) in 1487 non-dialysed patients across PEW stages 0 to 4]. Based on this classification, prevalence of PEW is 28% to 54% among 16 434 adults undergoing maintenance dialysis. PEW prevalence increases when renal function declines, that is, from <2% in CKD stages 1-2 to 11-54% in CKD stages 3-5. A more general definition of cachexia for all chronic diseases proposed by the Society on Sarcopenia, Cachexia and Wasting Disorders was also published concurrently. In the CKD area, we found 180 publications using 'cachexia' underlining that some confusion or overlap may exist. The definitions of PEW and cachexia are somewhat similar, and the main difference is that a loss of body weight >5% is a mandatory criterion for cachexia but supportive for PEW. The recent understanding of cachexia physiopathology during CKD progression suggests that PEW and cachexia are closely related and that PEW corresponds the initial state of a continuous process that leads to cachexia, implicating the same metabolic pathways as in other chronic diseases. Despite the success of the definition of PEW, using a more uniform term such as 'kidney disease cachexia' could be more helpful to design future research through collaborative groups of researchers with focus on cachexia.


TRH synthesis in "mute" thyrotropinomas: cause-effect or coincidence?
In the pathogenesis of thyrotropin (TSH) immunopositive pituitary adenomas, trigger mutagenetic events are well recognized. However, the way towards a clinical significant tumor is followed under the pressure of growth factors, among which the intrapituitary synthesis of releasing factors could bring a significant contribution. In this study, the production of thyrotropin releasing hormone (TRH) and beta TSH chain was evaluated at the mRNA level by in situ hybridization and end product level by immunohistochemistry, in 18 patients submitted to neurosurgery for pituitary macroadenomas. The hormonal sampling showed abnormal secretion for FSH in 5 and TSH in 4 patients. Seven cases were immunopositive for TSH, and expressed TSH beta mRNA. All but one out of these expressed also TRH mRNA. FSH immunoreactivity was documented in 12/ 18, only one of these being negative for TRH mRNA. Paracrine TRH could contribute to the pathogenesis of these "mute" adenomas.


Cerebral salt wasting syndrome following neurosurgical intervention in tuberculous meningitis.
Cerebral salt wasting is characterized by inappropriate natriuresis and volume contraction in the presence of cerebral pathology. Diagnosis can be difficult and therapy is challenging. We report two children with tuberculous meningitis and hydrocephalus who developed cerebral salt wasting following neurosurgical intervention. The first patient was managed with rigorous salt and water replacement whereas the second patient required the addition of fludrocortisone for control of salt-wasting.


Serum albumin and von Willebrand factor: possible markers for early detection of vascular damage in children undergoing peritoneal dialysis.
Cardiovascular diseases are the main causes of morbidity and mortality in children with end-stage renal disease and the relationships among several relevant potential biomarkers were investigated in pediatric peritoneal dialysis patients. Serum homocysteine, von Willebrand factor (vWF), apolipoproteins A and B, lipoprotein-a, high sensitive-CRP, hemoglobin, phosphorus and parathyroid hormone (PTH) levels, systolic (SBP) and diastolic (DBP) blood pressure, carotid intima-media thickness (cIMT) and left ventricular mass index (LVMI) were measured in 21 pediatric peritoneal dialysis patients and control subjects. All risk factors were higher in patients than controls. LVMI values were negatively correlated with hemoglobin and positively correlated with PTH and phosphorus levels (p=0.007, r= - 0.573; p=0.013, r= 0.532 and p=0.035, r= 0.461, respectively). cIMT was negatively associated with serum albumin and positively correlated with vWF levels and with SBP and DBP (p=0.006, r= - 0.578; p=0.039, r= 0.453; p=0.02, r= 0.503; p=0.024, r= 0.491, respectively). Robust regression analyses showed that hemoglobin was an independent predictor of LVMI and serum albumin was an independent predictor of cIMT. Only uremia-related factors were independent risk factors for predicting LVMI and cIMT. Hemoglobin level may be a critical factor in the development of left ventricular hypertrophy; therefore, effective treatment of anemia is crucial. Low serum albumin and high hsCRP and vWF levels, and their correlations with cIMT, indicate these patients could be at risk of developing malnutrition-inflammation-atherosclerosis syndrome and suggest that serum albumin and vWF levels may be useful markers for early detection of vascular damage.


[Loss of CDKN2 gene and amplification of cyclin D1 gene in human esophageal cancer].
The products of both CDKN2 and cyclin D1 genes are negative and positive regulators respectively in cell cycle. To study the involvement of the CDKN2 tumor suppressor gene and cyclin D1 oncogene in esophageal cancer development, we examined the situation of both genes in 21 pairs of primary human esophageal cancers and the mucosa adjacent to the cancers and also in four esophageal cancer cell lines by means of molecular biological and immunohistochemical techniques. Homozygous deletion was observed in 6 out of the 21 primary cancers and with lymph node metastasis in 5 out of the 6 cases. Loss of expression of p16 protein was identified immunohistochemically in 8 out of 21 primary cancers. Amplification of cyclin D1 was found in 12 out of 21 primary cancers and in 5 esophageal mucosa adjacent to the tumors, accompanying with overexpression of cyclin D1 protein. Homozygous deletion was observed in one (EC8712) out of the four cell lines, while by Northern and immunohisto chemistry analysis, loss of transcription of CDKN2 mRNA and loss of p16 protein expression were observed in two cell lines (EC8712 and EC8501). Amplification and overexpression of cyclin D1 were found in two cell lines (EC8733 and EC8501). These findings suggest that loss of CDKN2 gene and amplification of cyclin D1 gene are involved in esophageal cancers and that cyclin D1 alteration may be a earlier molecular event while CDKN2 to be a later one.


Analytical approach to the dynamics of facilitated spin models on random networks.
Facilitated spin models were introduced some decades ago to mimic systems characterized by a glass transition. Recent developments have shown that a class of facilitated spin models is also able to reproduce characteristic signatures of the structural relaxation properties of glass-forming liquids. While the equilibrium phase diagram of these models can be calculated analytically, the dynamics are usually investigated numerically. Here we propose a network-based approach, called approximate master equation (AME), to the dynamics of the Fredrickson-Andersen model. The approach correctly predicts the critical temperature at which the glass transition occurs. We also find excellent agreement between the theory and the numerical simulations for the transient regime, except in close proximity of the liquid-glass transition. Finally, we analytically characterize the critical clusters of the model and show that the departures between our AME approach and the Monte Carlo can be related to the large interface between blocked and unblocked spins at temperatures close to the glass transition.


Effects of rehydration on physiological and transcriptional responses of a water-stressed rhizobium.
As a microsymbiont of soybean, Bradyrhizobium japonicum plays an important role in symbiotic nitrogen fixation and sustainable agriculture. However, the survival of B. japonicum cells under water-deplete (e.g., drought) and water-replete (e.g., flood) conditions is a major concern affecting their nitrogen-fixing ability by establishing the symbiotic relationship with the host. In this study, we isolated a water stress tolerant rhizobium from soybean root nodules and tested its survival under water-deplete conditions. The rhizobium was identified as Bradyrhizobium japonicum and named strain 5038. Interestingly, both plate counting and live/dead fluorescence staining assays indicate that a number of viable but non-culturable cells exist in the culture medium upon the rehydration process which could cause dilution stress. Bradyrhizobium japonicum 5038 cells increased production of exopolysaccharide (EPS) and trehalose when dehydrated, suggesting that protective responses were stimulated. As expected, cells reduced their production upon the subsequent rehydration. To examine differential gene expression of B. japonicum 5038 when exposed to water-deplete and subsequent water-replete conditions, whole-genome transcriptional analysis was performed under 10% relative humidity (RH), and subsequent 100% RH, respectively. A total of 462 differentially expressed genes (DEGs, > 2.0-fold) were identified under the 10% RH condition, while 3,776 genes showed differential expression during the subsequent rehydration (100% RH) process. Genes involved in signal transduction, inorganic ion transport, energy production and metabolisms of carbohydrates, amino acids, and lipids were far more up-regulated than down-regulated in the 10% RH condition. Notably, trehalose biosynthetic genes (otsAB, treS, and treYZ), genes ligD, oprB, and a sigma factor rpoH were significantly induced by 10% RH. Under the subsequent 100% RH condition, genes involved in transcription, translation, cell membrane regulation, replication and repair, and protein processing were highly up-regulated. Interestingly, most of 10%-RH inducible genes displayed rehydration-repressed, except three genes encoding heat shock (Hsp20) proteins. Therefore, this study provides molecular evidence for the switch of gene expression of B. japonicum cells when encountered the opposite water availability from water-deplete to water-replete conditions.


Identification of 12(S)-hydroxyeicosatetraenoic acid in the young rat lens.
Evidence is presented indicating that young rat lens has the capacity to synthesize 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) from exogenous arachidonic acid (AA). A 9,000 xg supernatant prepared from 15 day old rat lenses, when incubated with calcium and U-[14C]-AA, generated a radiolabelled product with a retention time identical to authentic unlabelled 12-HETE in two different HPLC solvent systems. Mass spectral analysis provided evidence that the metabolite was 12-HETE while chiral studies demonstrated the exclusive presence of the 12(S) isomer. Radiolabelled 12-HETE synthesis was inhibited by preincubating the 15 day old rat lens supernatant with 0.2 Mm curcumin, a mixed cyclooxygenase/lipoxygenase inhibitor. Radiolabelled 12(S)-HETE synthetic capacity was highest in the 4 day old rat lens, the earliest time period measured, and rapidly declined with age reaching negligible levels at about 4 months. These results suggest that the young rat lens possess the biosynthetic capacity to generate radiolabelled 12(S)-HETE from U-[14]C-AA. The profile of 12-HETE synthetic activity suggests a possible regulatory function of the hydroxylated AA metabolite in the developing lens.


Visible Light-Based Ag3PO4/g-C3N4@MoS2 for Highly Efficient Degradation of 2-Amino-4-acetylaminoanisole (AMA) from Printing and Dyeing Wastewater.
In this research, the preparation of a Ag3PO4/g-C3N4@MoS2 photocatalyst and the performance and mechanism of degradation of 2-amino-4-acetaminoanisole (AMA) were studied. The phase composition and morphology of the synthesized samples were comprehensively characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), ultraviolet-visible diffuse reflectance (UV-Vis), and photoelectron spectroscopy (XPS). The catalytic performance of the photocatalyst was evaluated by the visible-light catalytic degradation of the AMA. The experimental results show that the Ag3PO4/g-C3N4@MoS2 composite photocatalyst has stronger photocatalytic oxidation and reduction capabilities than Ag3PO4 and Ag3PO4/g-C3N4. The material only decreases by 31.3% after five cycles of use, indicating that the material has good light stability. Free radical capture experiments prove that photo-generated holes (h+) and superoxide radicals (·O2-) are the main active substances in the photocatalytic process. The fundamental studies in the present research provide a new perspective for constructing an innovative type of visible-light photocatalyst and a new way to promote the photocatalytic degradation of organic pollutants.


Angiotensin-Converting Enzyme Genotype-Specific Immune Response Contributes to the Susceptibility of COVID-19: A Nested Case-Control Study.
Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of coronavirus disease 2019 (COVID-19), which has resulted in a global pandemic. Methodology: We used a two-step polymerase chain reaction to detect the ACE genotype and ELISA kits to detect the cytokine factor. We also used proteomics to identify the immune pathway related to the ACE protein expression. Result: In this study, we found that the angiotensin-converting enzyme (ACE) deletion polymorphism was associated with the susceptibility to COVID-19 in a risk-dependent manner among the Chinese population. D/D genotype distributions were higher in the COVID-19 disease group than in the control group (D/D odds ratio is 3.87 for mild (p value < 0.0001), 2.59 for moderate (p value = 0.0002), and 4.05 for severe symptoms (p value < 0.0001), logic regression analysis. Moreover, genotype-specific cytokine storms and immune responses were found enriched in patients with the ACE deletion polymorphism, suggesting the contribution to the susceptibility to COVID-19. Finally, we identified the immune pathway such as the complement system related to the ACE protein expression of patients by lung and plasma proteomics. Conclusion: Our results demonstrated that it is very important to consider gene polymorphisms in the population to discover a host-based COVID-19 vaccine and drug design for preventive and precision medicine.


Compatibility of Bacillus thuringiensis var. Israelensis and Bacillus sphaericus with the fungal pathogen Lagenidium giganteum (Oomycetes: Lagenidiales).
Larvae of Culex quinquefasciatus were exposed to infection by Lagenidium giganteum and various concentrations of B.t.i. or B. sphaericus. The resulting larval mortalities, percentages of infected dead larvae and percentages of larval body regions containing the fungus were compared. Overall, the effectiveness of Lagenidum giganteum against the larvae was not significantly affected by the presence of B.t.i. or B. sphaericus, and the fungal and bacterial agents were compatible. In experiments using 3-day-old larvae, the extent of growth of the fungus in the infected larvae and the percentage of the larvae infected were related to the concentration of B.t.i. in the range of 0.057-0.456 ITU/ml tested but were not related to the concentration of B. sphaericus in the range of 0.6-4.8 x 10(4) spores/ml. With larvae of various ages treated with a low concentration of B.t.i. (0.114 ITU/ml), exposure to the fungus increased the mortality rate in early but not late instars. After single and multiple applications of B.t.i. and B. sphaericus in the presence of the fungus, followed by drying and reflooding, the fungus persisted and reinfected larvae while the B. sphaericus persisted but the B.t.i. did not.


Effect of the unfolded protein response and oxidative stress on mutagenesis in CSF3R: a model for evolution of severe congenital neutropenia to myelodysplastic syndrome/acute myeloid leukemia.
Severe congenital neutropenia (SCN) is a rare blood disorder characterised by abnormally low levels of circulating neutrophils. The most common recurrent mutations that cause SCN involve neutrophil elastase (ELANE). The treatment of choice for SCN is the administration of granulocyte-colony stimulating factor (G-CSF), which increases the neutrophil number and improves the survival and quality of life. Long-term survival is however linked to the development of myelodysplastic syndrome/acute myeloid leukemia (MDS/AML). About 70% of MDS/AML patients acquire nonsense mutations affecting the cytoplasmic domain of CSF3R (the G-CSF receptor). About 70% of SCN patients with AML harbour additional mutations in RUNX1. We hypothesised that this coding region of CSF3R constitutes a hotspot vulnerable to mutations resulting from excessive oxidative stress or endoplasmic reticulum (ER) stress. We used the murine Ba/F3 cell line to measure the effect of induced oxidative or ER stress on the mutation rate in our hypothesised hotspot of the exogenous human CSF3R, the corresponding region in the endogenous Csf3r, and Runx1. Ba/F3 cells transduced with the cDNA for partial C-terminal of CSF3R fused in-frame with a green fluorescent protein (GFP) tag were subjected to stress-inducing treatment for 30 days (~51 doubling times). The amplicon-based targeted deep sequencing data for days 15 and 30 samples show that although there was increased mutagenesis observed in all the three genes of interest (partial CSF3R, Csf3r and Runx1), there were more mutations in the GFP region compared with the partial CSF3R region. Our findings also indicate that there is no correlation between the stress-inducing chemical treatments and mutagenesis in Ba/F3 cells. Our data suggest that oxidative or ER stress induction does not promote genomic instability, affecting partial C-terminal of the transduced CSF3R, the endogenous Csf3R and the endogenous Runx1 in Ba/F3 cells that could account for these targets to being mutational hotspots. We conclude that other mechanisms to acquire mutations of CSF3R that help drive the evolution of SCN to MDS/AML.


Policies Shaping Nursing Home Medication Practices: Involving Nurses to Advance Individualized Deprescribing.
Deprescribing is a common practice in the care of older adults, including those living in nursing homes (NHs). Deprescribing represents an individualized approach to optimizing medication use; it considers the risks, benefits, and goals of an individual, and can mitigate the effects of polypharmacy and potentially inappropriate medications. In NH settings, prescribing practices are shaped directly and indirectly by historical and contemporaneous policies at federal, state, and local levels, which have primarily targeted chemical restraints and unnecessary medications. Understanding these policies, their impact, and potentially unintended consequences is essential for gerontological nursing to transition toward individualized practices and approaches to deprescribing. [Journal of Gerontological Nursing, 49(9), 15-20.].


Use of personal EEG monitors in a behavioral neuroscience course to investigate natural setting sleep patterns and the factors affecting them in college students.
Sleep is often a topic of avid interest to college students, yet it is one that does not yield itself well to hands-on, interactive learning modules. Supplementing classroom learning with interactive "real world" laboratory activities provides students with a deeper understanding of behavior and its neural control. The project described here was designed to supplement the teaching of EEGs, sleep and circadian rhythms and involved students in the empirical process from hypothesizing about the factors that affect sleep, to personal data collection, data analysis and writing in the style of a peer-reviewed manuscript. Students enrolled in Behavioral Neuroscience at Connecticut College were provided with a home-based personal EEG monitor used to collect sleep data in their natural sleep setting. Participants recorded sleep data with the use of the ZEO® Personal Sleep Coach system and completed a nightly sleep journal questionnaire for seven nights. The ZEO® system uses EEG patterns to define sleep stages including wakefulness, light, deep and REM sleep. The journal included questions about factors known to affect sleep such as stress, caffeine, academic activity, exercise and alcohol. A class data set was compiled and used by students to perform univariate correlations examining the relationships between ZEO® variables and sleep journal variables. The data set allowed students to choose specific variables to investigate, analyze and write a peer-reviewed style manuscript. Significant class-wide correlations were found between specific sleep stages and behavioral variables suggesting that the ZEO® system is sophisticated yet inexpensive enough to be used as an effective tool in the classroom setting. Overall student feedback on the exercise was positive with many students indicating that it significantly enhanced their understanding of sleep architecture and made them keenly aware of the factors that affect quality of sleep.


The buttonhole technique: strategies to reduce infections.
The buttonhole technique has gained popularity over the last decade in the United States. The fact that it is a relatively new technique, which requires a change to current cannulation practice, has contributed to several unforeseen complications that have led to increased infection rates in AV fistulae. To keep this technique a viable option for patients, it will be necessary to understand the potential infection risks and implement strategies to reduce the incidence of infection.


Influence of hypothermia and cardioplegic solutions on expression of alpha-Gal epitope on porcine aortic endothelial cells.
The Galalpha1-3Galbeta1-4GlcNAc-R is the major antigen on pig tissue bound by human xenoreactive natural antibodies in xenotransplant. We have investigated in vitro the influence of hypothermic storage with cardioplegic solutions on expression of Galalpha1-3Galbeta1-4GlcNAc-Rs and hyperacute xenograft rejection. To analyze effects of hypothermia on the Galalpha1-3Galbeta1-4GlcNAc-Rs, cultured porcine aortic endothelial cells were exposed to a temperature of 4 degrees C for 1 hour, 4 hours, and 6 hours. Cell cultures of the control groups were incubated at the same time at 38 degrees C. To investigate the influence of cardioplegic solutions on the Galalpha1- 3Galbeta1-4GlcNAc-Rs, porcine aortic endothelial cells were exposed to 4 degrees C for 4 hours in the presence of University of Wisconsin solution or histidinetryptophan- ketoglutarate solution. Cells of the control groups were cooled at 4 degrees C for 4 hours without cardioplegic solution. After treatment, porcine aortic endothelial cells were submitted to fluorescence-activated cell sorter. Hypothermia of 4 degrees C showed no significant effect on the quantity of Galalpha1-3Galbeta1-4GlcNAc-Rs. However, the treatment of porcine aortic endothelial cells with University of Wisconsin solution resulted in a highly significant reduction of Galalpha1-3Galbeta1- 4GlcNAc-Rs by 50% (P = .006). Treatment of porcine aortic endothelial cells with histidine-tryptophanketoglutarate solution decreased Alpha-Gal quantity significantly by 32% (P = .011). Our data offer new perspectives in the prevention of hyperacute, humoral xenograft rejection by reducing the Galalpha1-3Galbeta1-4GlcNAc-Rs after exposure to different cardioplegic solutions.


Effect of Cyclic Loading on Surface Instability of Silicone Rubber under Compression.
This work combines experiments and finite element simulations to study the effect of pre-imposed cyclic loading on surface instability of silicon rubber under compression. We first fabricate cuboid blocks of silicon rubber and pinch them cyclicly a few times. Then, an in-house apparatus is set to apply uniaxial compression on the silicon rubber under exact plane strain conditions. Surprisingly, we find multiple creases on the surface of silicone rubber, significantly different from what have been observed on the samples without the cyclic pinching. To reveal the underlying physics for these experimentally observed multiple creases, we perform detailed nanoindentation experiments to measure the material properties at different locations of the silicon rubber. The modulus is found to be nonuniform and varies along the thickness direction after the cyclic pinching. According to these experimental results, three-layer and multilayer finite element models are built with different materials properties informed by experiments. The three-layer finite element model can excellently explain the nucleation and pattern of multiple surface creases on the surface of compressed silicone rubber, in good agreement with experiments. Counterintuitively, the multilayer model with gradient modulus cannot be used to explain the multiple creases observed in our experiments. According to these simulations, the experimentally observed multiple creases should be attributed to a thin and stiff layer formed on the surface of silicon rubber after the pre-imposed cyclic loading.


Discovery and characterization of de novo sialic acid biosynthesis in the phylum Fusobacterium.
Sialic acids are nine-carbon backbone carbohydrates found in prominent outermost positions of glycosylated molecules in mammals. Mimicry of sialic acid (N-acetylneuraminic acid, Neu5Ac) enables some pathogenic bacteria to evade host defenses. Fusobacterium nucleatum is a ubiquitous oral bacterium also linked with invasive infections throughout the body. We employed multidisciplinary approaches to test predictions that F. nucleatum engages in de novo synthesis of sialic acids. Here we show that F. nucleatum sbsp. polymorphum ATCC10953 NeuB (putative Neu5Ac synthase) restores Neu5Ac synthesis to an Escherichia coli neuB mutant. Moreover, purified F. nucleatum NeuB participated in synthesis of Neu5Ac from N-acetylmannosamine and phosphoenolpyruvate in vitro Further studies support the interpretation that F. nucleatum ATCC10953 NeuA encodes a functional CMP-sialic acid synthetase and suggest that it may also contain a C-terminal sialic acid O-acetylesterase. We also performed BLAST queries of F. nucleatum genomes, revealing that only 4/31 strains encode a complete pathway for de novo Neu5Ac synthesis. Biochemical studies including mass spectrometry were consistent with the bioinformatic predictions, showing that F. nucleatum ATCC10953 synthesizes high levels of Neu5Ac, whereas ATCC23726 and ATCC25586 do not express detectable levels above background. While there are a number of examples of sialic acid mimicry in other phyla, these experiments provide the first biochemical and genetic evidence that a member of the phylum Fusobacterium can engage in de novo Neu5Ac synthesis.


US regional health information organizations and the nationwide health information network: any lessons for Canadians?
The creation of regional clinical data exchanges (usually referred to as RHIOs) is a centerpiece of the US national healthcare information technology strategy. How well are they doing and what lessons can we learn that might be applied here in Canada?


[The expression of various cytokeratins by epithelial cells of calcified odontogenic cysts].
The calcifying odontogenic cyst (COC) is a rare lesion without specific clinical characteristics. Its diagnosis is essentially histopathological showing the presence of ghost cells associated with narrow squamous epithelium, the basal strata of which consist of clearly delineated cells differentiating in areas of the stellate reticulum in a similar way to ameloblastoma. The results of ultrastructural observations of the ghost cells as well as histochemical and immunohistochemical studies suggest that they are the sites of abnormal keratinisation. The aim of this study of one COC mas to locate the stages in epithelial maturation associated with the formation of ghost cells. Six monoclonal antibodies were used; three with a wide spectrum (KL1, AE3, AE1), two with a narrow spectrum against high molecular weight cytokeratins (AE2, AE8) and one against vimentin (M725). Histopathological examination of the COC revealed three different types of cells in the epithelial lining and the epithelial islands; small basal cubic cells surrounding larger ones placed centrally or suprabasally; balloon shaped cells or flattened ghost cells rolled up on themselves to resemble keratinising pearls, or cornifying cells. The ghost cells and cornifying cells had an altered distribution of their cytokeratins demonstrated by the absence of staining of antibodies against cytokeratins. The differentiated cells adjacent to them showed cytokeratins typical of squamous epithelium rather than those associated with the process of keratinization. The coexistence of cornifying and ghost cells testifies to the great potential of odontogenic epithelium to form numerous epithelial islands.


Infants' ability to parse continuous actions: further evidence.
In two experiments, we examined 6- and 8-month-old infants' capacities to detect target actions in a continuous action sequence. The primary question was whether action segments consisting of an event (e.g., occlusion, containment) are more salient than action segments consisting of a transition (e.g., bounce, slide). In Experiment 1, infants were habituated to long action sequences. After meeting the habituation criterion, infants were shown an alternation between test trials consisting of either novel or familiar segments made up of an event and transition. The results demonstrate that infants dishabituated to the novel test segments. In Experiment 2, infants were habituated to the same long action sequences but the novelty/familiarity of the events and transitions were crossed with each other. The results demonstrate that infants looked longer at test trials with novel events compared to test trials with novel transitions. These experiments replicated and extended the phenomena reported in Hespos, Saylor, and Grossman (2009). Together these findings demonstrated that in event processing, events having greater relative salience than transitions. These findings suggest that object knowledge could provide insights to the process of event segmentation.


In vitro and in vivo photodynamic therapy of otitis media in gerbils.
The aim of this study was to evaluate the antibacterial effects of photodynamic therapy (PDT) on common bacteria causing otitis media with effusion (OME). An in vitro study was carried out using a hematoporphyrin derivative sensitizer (Photogem; Lemonosov Institute of Fine Chemical, Moscow, Russia) and a 632-nm diode laser on Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. The presence of colony-forming units of the bacteria was examined, the microscopic structures of the bacteria were examined by transmission electron microscopy (TEM), and flow cytometry of the bacteria was performed. An in vivo PDT study was performed using gerbils. S. pneumoniae or H. influenzae were injected into bullae. The Photogem was injected into the bullae 2 days later when OME developed, and transcanal irradiation with the 632-nm diode laser (90 J) was performed. Middle ear and bulla were washed with Dulbecco's phosphate buffered saline (DPBS) and the washed DPBS was cultured. The presence of bacterial colonies was examined. The PDT was effective in killing all three kinds of bacteria. TEM showed damaged bacterial cell membranes and cytoplasmic structures, and the flow cytometry showed a lower number of viable bacteria in the PDT group compared to the control group. PDT was effective in killing S. pneumoniae in 87% of the infected bullae with OME, whereas it was effective in eradicating H. influenzae in 50% of the infected bullae with OME. The results of these studies demonstrated that PDT may be effective to treat otitis media. PDT may have clinical implications in the treatment of otitis media that is resistant to antibiotic therapy.


Abnormal fragile histidine triad (FHIT) expression in advanced cervical carcinoma: a poor prognostic factor.
The FHIT gene is a candidate tumor suppressor gene that has been implicated in the development of cervical carcinoma. We hypothesized that abnormal Fhit expression might be a poor prognostic factor for patients with cervical cancer. The tumors from 59 high-risk patients (stage II-III) were evaluated for abnormal Fhit expression by immunohistochemical staining. Abnormal Fhit expression (absent or reduced) was noted in 66% of the specimens. There was no statistical difference with respect to stage, performance status, para-aortic node metastasis, completion of therapy, grade, race, age, and HIV status between the normal and abnormal Fhit expression groups. The 3-year survival for patients whose tumors displayed normal Fhit expression versus abnormal Fhit expression was 74% versus 37%, respectively. Univariate analysis demonstrated a difference in survival that was statistically significant for age <55 years versus > or =55 years (P = 0.015), normal Fhit expression versus abnormal Fhit expression (P = 0.015), and stage II versus stage III (P = 0.033). Multivariate analysis showed that abnormal Fhit expression was a poor prognostic factor (P = 0.015).


Lipofuscin accumulation in neurons with restraint stress.
Stress induced lipofuscinosis was studied in rat cerebellum. 3 month old wistar rats were subjected to restraint stress by keeping them immobile for 24, 48 and 72 hours duration. This was achieved in specially prepared cages which allowed no space for the rats to move; giving a stress to the animal. The cerebella from the stressed groups rats were removed after the experiment and were processed for fluorescent microscopical, histochemical and fluorimetric study of lipofuscin. The lipofuscin content in the Purkinje neurons was compared with that of the control rats which were of the same age, size and weight as of the experimental rats. The results showed that the lipofuscin content in the neurons of the experimental rats was more than that of the control ones. In gist, while 24 hrs. stress caused a 28.9% increase in lipofuscin content, 48 hrs. stress resulted in a 38.3% increase. This shows that restraint stress can be a good experimental model for lipofuscinogenesis and ageing studies.


Hand Hygiene Compliance at Two Tertiary Hospitals in Freetown, Sierra Leone, in 2021: A Cross-Sectional Study.
Hand hygiene actions are essential to reduce healthcare-associated infections and the development of antimicrobial resistance. In this cross-sectional study at two tertiary hospitals, Freetown, Sierra Leone, we observed hand hygiene compliance (defined as using handwash with soap and water or alcohol-based hand rub (ABHR) amongst healthcare workers between June and August 2021. Using the WHO Hand Hygiene tool, observations were made in relation to the type of opportunity, different wards and types of healthcare worker. Overall, 10,461 opportunities for hand hygiene were observed, of which 5086 (49%) resulted in hand hygiene actions. ABHR was used more often than handwash (26% versus 23%, p < 0.001). Overall, compliance was significantly better: after being with a patient/doing a procedure than before (78% after body fluid exposure risk compared with 24% before touching a patient—p < 0.001); in Paediatric (61%) compared with Medical wards (46%)—p < 0.001; and amongst nurses (52%) compared with doctors (44%)—p < 0.001. Similar patterns of compliance were observed within each hospital. In summary, hand hygiene compliance was sub-optimal, especially before being with a patient or before clean/aseptic procedures. Improvement is needed through locally adapted training, hand hygiene reminders in wards and outpatient departments, uninterrupted provision of ABHR and innovative ways to change behaviour.


Validation of ultrasound imaging for Achilles entheseal fibrocartilage in bovines and description of changes in humans with spondyloarthritis.
Entheseal fibrocartilage (EF) derangement is hypothesised to be pivotal to the pathogenesis of spondyloarthritis. Ultrasound is useful for visualisation of the enthesis but its role in EF visualisation is uncertain. This work aimed to demonstrate face and content validity of ultrasound for EF visualisation both by bovine histological evaluation and EF imaging in spondyloarthritis. Achilles enthesis of 18 bovine hindfeet was visualised using a MyLab 70 ultrasound machine. The presence of tissue with EF characteristics was documented and histological confirmation was performed on five randomly selected sections using Masson trichrome staining. Ultrasound of the Achilles tendon (AT) was performed in 19 patients with spondyloarthritis and 21 healthy controls (HC). The bovine EF could be visualised in all cases and seen as a thin, uncompressible, well-defined, anechoic layer between the hyperechoic bone and the hyperechoic fibrils of the enthesis both in longitudinal and transverse scans. This region corresponded to EF on histological examination. The same pattern of low signal corresponding to EF location was seen in 17/19 patients and all HC. Discontinuities of the anechoic layer around the erosions and enthesophytes were observed in the spondyloarthritis group. The thickness of the anechoic layer was not significantly different in spondyloarthritis and HC (0.5 ± 0.1 vs 0.5 ± 0.2 mm, p=0.9) whereas the thickness of the EF was greater in men (0.6 ± 0.2 vs 0.5 ± 0.1 mm; p=0.009) compared with women. Ultrasound can visualise EF of the AT insertion, which can be abnormal in cases of spondyloarthritis. This has implications for a better understanding of enthesopathy.


Isolation and characterization of an extracellular antimicrobial protein from Aspergillus oryzae.
A 17 kDa antimicrobial protein was isolated from growth medium containing the filamentous fungus Aspergillus oryzae by extracting the supernatants from the culture media, ion exchange chromatography on CM-sepharose, and C18 reverse-phase high-performance liquid chromatography. This antimicrobial protein, which we considered to be an extracellular antimicrobial protein from A. oryzae (exAP-AO17), possessed antimicrobial activity but lacked hemolytic activity. The exAP-AO17 protein strongly inhibited pathogenic microbial strains, including pathogenic fungi, Fusarium moniliform var. subglutinans and Colletotrichum coccodes, and showed antibacterial activity against bacteria, including E. coli O157 and Staphylococcus aureus. To confirm that the protein acts as a regulation factor for extracellular secretion, we examined growth under varying conditions of N sources, C sources, ions, ambient pH, and stress. Various culture conditions were found to induce characteristic changes in the expression of protein synthesis as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Highly basic polypeptides were regulated by suppressing the ambient pH under acidic conditions and strongly induced under alkaline conditions, thus confirming that pH regulation is physiologically relevant. The expression of exAP-AO17 was upregulated by heat shock upon growth in the presence of NaCl. Automated Edman degradation showed that the N-terminal sequence of exAP-AO17 was NH 2-GLPGPAGAVGFAGKDQNM-. ExAP-AO17 showed partial sequence homology with a collagen belonging to the animal source. These results suggest that exAP-AO17 is an excellent candidate as a lead compound for the development of novel oral or other types of anti-infective agents.


C3 Vertebral Metastases From Tongue Adenoid Cystic Carcinoma: A Rare Case Report.
We report a rare case involving a patient with C3 vertebral body metastasis secondary to adenoid cystic carcinoma of the tongue.Five years after local resection of the primary tumor, magnetic resonance imaging showed a metastasis located in the left posterior border of the C3 vertebral body. Additionally, multiple pulmonary metastases were identified by computed tomography. Based on these findings, the patient underwent C2-3, C3-4 discectomy; C3 corpectomy; and titanium mesh fusion with a Zephir plate. The diagnosis was confirmed by the pathology findings. During 6 months of follow-up, the patient showed improvement and return of function of the cervical vertebrae, with no serious complications.Because of the scarcity of cases of vertebral metastases from tumors of the tongue in the literature, we have reported this case to add to the available evidence regarding this rarely encountered condition.


Determination of cranio-spinal canal compliance distribution by MRI: Methodology and early application in idiopathic intracranial hypertension.
To develop a method for derivation of the cranial-spinal compliance distribution, assess its reliability, and apply to obese female patients with a diagnosis of idiopathic intracranial hypertension (IIH). Phase contrast-based measurements of blood and cerebrospinal fluid (CSF) flows to, from, and between the cranial and spinal canal compartments were used with lumped-parameter modeling to estimate systolic volume and pressure changes from which cranial and spinal compliance indices are obtained. The proposed MRI indices are analogous to pressure volume indices (PVI) currently being measured invasively with infusion-based techniques. The consistency of the proposed method was assessed using MRI data from seven aged healthy subjects. Measurement reproducibility was assessed using five repeated MR scans from one subject. The method was then applied to compare spinal canal compliance contribution in seven IIH patients and six matched healthy controls. In the healthy subjects, as expected, spinal canal contribution was consistently larger than the cranial contribution (average value of 69%). Measurement variability was 8%. In IIH, the spinal canal contribution is significantly smaller than normal controls (60 versus 78%, P < 0.03). An MRI-based method for derivation of compliance indices analogous to PVI has been implemented and applied to healthy subjects. The application of the method to obese IIH patients suggests a spinal canal involvement in the pathophysiology of IIH.


Perspectives on clinical outcomes assessment: a view for the allied health professions.
The purpose of clinical outcomes assessment is well recognized by allied health professionals, but several obstacles have impeded the conduct of such research in the allied health professions. Many professionals have difficulty in determining how to begin. To explore the research possibilities already available within the health care environment, a way to visualize and organize the potential variables is needed. In light of a taxonomy of patient outcomes, an effort is made here to summarize and portray the characteristics of the delivery system, the environmental and structural factors, and both patient and provider attributes. Many of these aspects of outcomes investigations are within the purview and capabilities of allied health professionals. Through a division of labor among the professions, a mosaic can be collectively created that validates what they do and the value they add to patient care.


Outcomes and complications of open, laparoscopic, and hybrid giant ventral hernia repair.
An incisional hernia is a common complication of abdominal surgery. To evaluate the outcomes and complications of hybrid application of open and laparoscopic approaches in giant ventral hernia repair. Medical records of patients who underwent open, laparoscopic, or hybrid surgery for a giant ventral hernia from 2006 to 2013 were retrospectively reviewed. The hernia recurrence rate and intra- and postoperative complications were calculated and recorded. Open, laparoscopic, and hybrid approaches were performed in 82, 94, and 132 patients, respectively. The mean hernia diameter was 13.11 ± 3.4 cm. The incidence of hernia recurrence in the hybrid procedure group was 1.3%, with a mean follow-up of 41 mo. This finding was significantly lower than that in the laparoscopic (12.3%) or open procedure groups (8.5%; P < 0.05). The incidence of intraoperative intestinal injury was 6.1%, 4.1%, and 1.5% in the open, laparoscopic, and hybrid procedures, respectively (hybrid vs open and laparoscopic procedures; P < 0.05). The proportion of postoperative intestinal fistula formation in the open, laparoscopic, and hybrid approach groups was 2.4%, 6.8%, and 3.3%, respectively (P > 0.05). A hybrid application of open and laparoscopic approaches was more effective and safer for repairing a giant ventral hernia than a single open or laparoscopic procedure.


Proteomimetic libraries: design, synthesis, and evaluation of p53-MDM2 interaction inhibitors.
The p53-MDM2 interaction regulates p53-mediated cellular responses to DNA damage, and MDM2 is overexpressed in 7% of all cancers. Structure-based computational design was applied to this system to design libraries centered on a scaffold that projects side chain functionalities with distance and angular relationships equivalent to those seen in the MDM2 interacting motif of p53. A library of 173 such compounds was synthesized using solution phase parallel chemistry. The in vitro competitive ability of the compounds to block p53 peptide binding to MDM2 was determined using a fluorescence polarization competition assay. The most active compound bound with K(d) = 12 microM, and its binding was characterized by (15)N-(1)H HSQC NMR.


Textile effluent induced genotoxic effects and oxidative stress in Clarias gariepinus.
Human and ecological disorder experienced in industrial settlements as a result of improper disposal of chemicals such as textile effluent calls for careful surveillance on the state of the environment. This study investigated the toxicity of textile effluent discharge using biochemical and cytogenetic responses to ascertain the acute and sub lethal effects on Clarias gariepinus. The 96 h LC50 of C. gariepinus exposed to the textile effluent was 8.203 ml L(-1). Fourteen day exposures to 1, 2, 4 and 6 ml L(-1) doses were conducted and several toxicological endpoints were evaluated. Sub lethal genotoxicity and biochemical study was also carried out for fourteen days. The genotoxicity studies utilized micronucleus test while the biochemical studies quantified serum anti-oxidant status Total Protein (TP), Catalase (CAT), Superoxide Dismutase (SOD) and Malondialdehyde (MDA) of the exposed fish. Toxicity factor indicates that the 96 h LC50 was significantly more toxic than the 24 h LC50 (p < 0.05). The textile effluent at the tested concentrations induced micronucleus and nuclear abnormalities in the peripheral blood of exposed fish. Micronucleus, notch and binucleated cell formation were significant (p < 0.05) compared to control while lobed and blebbed cells were insignificant (p < 0.05). SOD, TP and CAT significantly (p < 0.05) decreased compared to control group while MDA increased compared to control but was insignificant (p > 0.05). The results obtained from this study showed that textile effluent increase cytogenetic damage and altered anti-oxidant status in C. gariepinus. Chemicals in the effluent can be bioaccumulated and biomagnified in the aquatic organism hence affecting man.


The Antibacterial Activity of Erythrocytes From Goose (Anser domesticus) Can Be Associated With Phagocytosis and Respiratory Burst Generation.
In the lumen of blood vessels, there are large numbers of erythrocytes, which are approximately 95% of the total blood cells. Although the function of erythrocytes is to transport oxygen in the organism, recent studies have shown that mammalian and teleost erythrocytes are involved in the immune response against bacterial infections. However, the immune mechanisms used by avian erythrocytes are not yet clear. Here, we demonstrated that erythrocytes from goose have the ability to phagocytose as well as conduct antimicrobial activity. Firstly, we revealed the phagocytosis or adhesion activity of goose erythrocytes for latex beads 0.1-1.0 μm in diameter by fluorescence microscopy, and scanning and transmission electron microscopy. The low cytometry results also proved that goose erythrocytes had a wide range of phagocytic or adhesion activity for different bacteria. Followed, the low cytometry analysis data further explored that the goose erythrocytes contain the ability to produce reactive oxygen species (ROS) and inducible nitric oxide synthase (iNOS) in response to bacterial stimulation, and also up-regulated the expression of NOX family includes NOX1 and NOX5. Finally, we also found that goose erythrocytes showed a powerful antibacterial activity against all the three bacteria, meanwhile the stimulation of three kinds of bacteria up-regulated the expression of inflammatory factors, and increased the production of antioxidant enzymes to protect the cells from oxidative damage. Herein, our results demonstrate that goose Erythrocytes possess a certain phagocytic capacity and antioxidant system, and that the antimicrobial activity of erythrocytes can occurred through the production of unique respiratory burst against foreign pathogenic bacteria, which provides new clues to the interaction between bacteria and avian erythrocytes.


An MR image-guided, voxel-based partial volume correction method for PET images.
Partial volume effect in positron emission tomography (PET) can cause incorrect quantification of radiopharmaceutical uptake in functional imaging. A PET partial volume correction method is presented to attenuate partial volume blurring and to yield voxel-based corrected PET images. By modeling partial volume effect as a convolution of point spread function of the PET scanner, the reconstructed PET images are corrected by iterative deconvolution with an edge-preserving smoothness constraint. The constraint is constructed to restore discontinuities extracted from coregistered MR images but maintains the smoothness in radioactivity distribution. The correction is implemented in a Bayesian deconvolution framework and is solved by a conjugate gradient method. The performance of the method was compared with the geometric transfer matrix (GTM) method on a simulated dataset. The method was evaluated on synthesized brain FDG-PET data and phantom MRI-PET experiments. The true PET activity of objects with a size of greater than the full-width at half maximum of the point spread function has been effectively restored in the simulated data. The partial volume correction method is quantitatively comparable to the GTM method. For synthesized FDG-PET with true activity 0 μci/cc for cerebrospinal fluid (CSF), 228 μci/cc for white matter (WM), and 621 μci/cc for gray matter (GM), the method has improved the radioactivity quantification from 186 ± 16 μci/cc to 30 ± 7 μci/cc in CSF, 317 ± 15 μci/cc to 236 ± 10 μci/cc for WM, 438 ± 4 μci/cc to 592 ± 5 μci/cc for GM. Both visual and quantitative assessments show improvement of partial volume correction in the synthesized and phantom experiments. The partial volume correction method improves the quantification of PET images. The method is comparable to the GTM method but does not need MR image segmentation or prior tracer distribution information. The voxel-based method can be particularly useful for combined PET/MRI studies.


Acute leukemia in children: A review of the current Indian data.
Acute leukemias are the most common diagnostic group of childhood cancer. This review summarizes the published literature on reported current outcomes of childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) from India. Overall survival in ALL ranged from 45% to 81% (commonly >60%) and event-free survival ranged from 41% to 70% (commonly >50%). Outcome data for AML was patchy with varying duration of follow-up, but it can be inferred that 50-80% of treated patients had experienced an event (toxic death, refractory disease or relapse). It is imperative that going forward focus should be on collaborative efforts, which promote treatment of patients on risk-stratified adapted protocols based on local infrastructure, improvement in supportive care and encourage prospective multi-center clinical trials.


Role of surgery in treatment of locally advanced prostate cancer.
The proportion of prostate cancer diagnosed at localized stages increased from 56.7% to 74. 0% between 1973 and 1993 ("stage migration"). A corresponding increase in the number of radical prostatectomies performed each year was also noted. Nomograms are mathematical algorithms derived from statistical models that are used to predict outcomes for an individual patient, or for groups of patients. In fact, careful pre-operative patient and tumor selection before radical prostatectomy is mandatory. Locally advanced prostate cancer is defined as tumor that has extended clinically beyond the prostatic capsule, with invasion of the pericapsular tissue, apex, bladder neck or seminal vescicle, but without lymph node involvement or distant metastasis. It is estimated that 12-15% of prostate cancer are stage T3. Overstaging or understaging of this cancer is common. Correct staging of clinical T3 disease is even more difficult and both overstaging pT2 and understaging pT4 or pN+ are common. The goals of treatment for T3 tumors are to cure the disease, prolong survival or metastasis-free survival and improve the quality of life. The authors reviewed the most important studies, investigated radical prostatectomy as monotherapy for locally advanced prostate cancer and the integration of surgery with hormonal treatment. The EAU guidelines on prostate cancer state that radical prostatectomy in locally advanced disease is an option for selected patients with small T3, PSA <20 ng/ml, Gleason score <8 and a life expectancy > 10 years. Ten to 15% of clinical T3 are overstaged as pT2. This may lead to the possibility of curing these patients with surgery as the monotherapy. The increased use of nomograms and increased knowledge of recognized prognostic factors could lead to the selection of a large number of patients, often with a long life expectancy, who could benefit from surgical treatment.


Effects of Pseudomonas aeruginosa endotoxin on vasodilation in the intact spinotrapezius muscle.
The purpose of this study was to determine whether short-term exposure to clinically relevant concentrations of Pseudomonas aeruginosa lipopolysaccharide (LPS) impairs vasoreactivity of resistance arterioles in the intact spinotrapezius muscle microcirculation and, if so, to determine the mechanisms mediating this response. Using intravital microscopy, we found that 60-min suffusion of P. aeruginosa LPS (0.03-3.0 microg/ml) on the in situ hamster spinotrapezius muscle elicited an immediate, profound, and prolonged concentration-dependent vasodilation (P < 0.05). This response was reversible once suffusion of P. aeruginosa LPS was stopped. Pretreatment with N(G)-nitro-L-arginine methyl ester (10.0 microM), a nonselective nitric oxide (NO) synthase inhibitor, but not N(G)-nitro-D-arginine methyl ester, abrogated P. aeruginosa LPS-induced vasodilation and elicited a small, albeit significant, vasoconstriction. Indomethacin had no significant effects on P. aeruginosa LPS-induced responses. P. aeruginosa LPS had no significant effects on acetylcholine- and nitroglycerin-induced vasodilation in the spinotrapezius muscle. Collectively, these data indicate that short-term exposure to clinically relevant concentrations of P. aeruginosa LPS evokes an immediate, potent, prolonged, and reversible NO-dependent, prostaglandin-independent vasodilation in skeletal muscles in vivo. We suggest this response could play an important role in the pathophysiology of the profound vasomotor dysfunction observed in the peripheral circulation of patients with P. aeruginosa sepsis syndrome.


Molecular cloning, expression pattern, and chromosomal localization of the human Na-Cl thiazide-sensitive cotransporter (SLC12A3).
Electrolyte homeostasis is maintained by several ion transport systems. Na-(K)-Cl cotransporters promote the electrically silent movement of chloride across the membrane in absorptive and secretory epithelia. Two kidney-specific Na-(K)-Cl cotransporter isoforms are known, so far, according to their sensitivity to specific inhibitors. We have cloned the human cDNA coding for the renal Na-Cl cotransporter selectively inhibited by the thiazide class of diuretic agents. The predicted protein sequence of 1021 amino acids (112 kDa) shows a structure common to the other members of the Na-(K)-Cl cotransporter family: a central region harboring 12 transmembrane domains and the 2 intracellular hydrophilic amino and carboxyl termini. The expression pattern of the human Na-Cl thiazide-sensitive cotransporter (hTSC, HGMW-approved symbol SLC12A3) confirms the kidney specificity. hTSC has been mapped to human chromosome 16q13 by fluorescence in situ hybridization. The cloning and characterization of hTSC now render it possible to study the involvement of this cotransport system in the pathogenesis of tubulopathies such as Gitelman syndrome.


Esomeprazole inhibits hypoxia/endothelial dysfunction-induced autophagy in preeclampsia.
Preeclampsia (PE) affects 3 to 5% of pregnant women worldwide and is associated with fetal and maternal morbidity and mortality. Although a complete understanding of PE remains elusive, it has been widely accepted that a dysfunction of the placenta plays a key role in the pathogenesis of PE. In this study, we investigated the role of excessive placental autophagy during PE pathogenesis and explored whether esomeprazole ameliorates PE by inhibiting the autophagy in the placenta. The PE cellular model was established by treating the cells' L-NAME and hypoxia. The PE mice model was established by L-NAME administration and was confirmed by the increased systolic blood pressure (SBP) and urinary protein detected. The autophagy and key proteins were detected in human placental tissue, in cells, and in the mice model by Western blot and immunofluorescence staining. Results showed that excessive autophagy could be detected in human PE placental tissue, in the PE cellular model, and in the PE mice model. Hypoxia induces autophagy by activating AMPKα and inhibiting mTOR in vivo and in vitro. Esomeprazole inhibits L-NAME-induced autophagy in mice by inhibiting AMPKα and activating mTOR. In conclusion, this study demonstrates that the excessive autophagy induced by the SIRT1/AMPKα-mTOR pathway plays a significant role in the pathogenesis of PE. However, esomeprazole treatment inhibits AMPKα but activates mTOR, resulting in the inhibition of autophagy in the placenta and, therefore, mitigates PE symptoms.


Breaking the integrin hinge. A defined structural constraint regulates integrin signaling.
Integrins are heterodimeric (alpha, beta) cell adhesion receptors. We demonstrate that point mutations in the cytoplasmic domains of both the alpha and beta subunits promote constitutive signaling by the integrin alphaIIbbeta3. By generating charge reversal mutations, we show these "activating" mutations may act by disrupting a potential salt bridge between the membrane-proximal portions of the alpha and beta subunit cytoplasmic domains. Thus, the modulation of specific interactions between the alpha and beta subunit cytoplasmic domains may regulate transmembrane signaling through integrins. In addition, these activating mutations induce dominant alterations in cellular behavior, such as the assembly of the extracellular matrix. Consequently, somatic mutations in integrin cytoplasmic domains could have profound effects in vivo on integrin-dependent functions such as matrix assembly, cell migration, and anchorage-dependent cell growth and survival.


Microbiological diagnosis of sepsis: the confounding effects of a "gold standard".
The need of rapid and sensitive diagnostic techniques for sepsis is every day more compelling. Its morbidity and mortality loads are dramatically high, with one quarter of patients eventually dying. Several diagnostic progresses have been made in the last years using both molecular- and nonmolecular-based approaches, and they have to be broadly shared in the scientific community also under the technical point of view. The initial chapters of this book give a thorough overlook of the state of the art in the actual diagnosis of sepsis. The other chapters provide a broad range of protocols describing both already used and futuristic tools, covering both microbiological and nonmicrobiological aspects. The potential role of each described protocol is evidenced by a brief introduction on the specific topic of each chapter. A final chapter describing algorithms potentially useful in stratifying the risk of sepsis in each single patient and suggesting the future perspectives in the diagnosis of sepsis closes the book.