[Non-beneficial therapy and emotional exhaustion in end-of-life care : Results of a survey among intensive care unit personnel].
End-of-life care (EOLC) in the intensive care unit (ICU) is becoming increasingly more common but ethical standards are compromised by growing economic pressure. It was previously found that perception of non-beneficial treatment (NBT) was independently associated with the core burnout dimension of emotional exhaustion. It is unknown whether factors of the work environment also play a role in the context of EOLC. Is the working environment associated with perception of NBT or clinician burnout? Physicians and nursing personnel from 11 German ICUs who took part in an international, longitudinal prospective observational study on EOLC in 2015-2016 were surveyed using validated instruments. Risk factors were obtained by multivariate multilevel analysis. The participation rate was 49.8% of personnel working in the ICU at the time of the survey. Overall, 325 nursing personnel, 91 residents and 26 consulting physicians participated. Nurses perceived NBT more frequently than physicians. Predictors for the perception of NBT were profession, collaboration in the EOLC context, excessively high workload (each p ≤ 0.001) and the numbers of weekend working days per month (p = 0.012). Protective factors against burnout included intensive care specialization (p = 0.001) and emotional support within the team (p ≤ 0.001), while emotional exhaustion through contact with relatives at the end of life and a high workload were both increased (each p ≤ 0.001). Using the example of EOLC, deficits in the work environment and stress factors were uncovered. Factors of the work environment are associated with perceived NBT. To reduce NBT and burnout, the quality of the work environment should be improved and intensive care specialization and emotional support within the team enhanced. Interprofessional decision-making among the ICU team and interprofessional collaboration should be improved by regular joint rounds and interprofessional case discussions. Mitigating stressful factors such as communication with relatives and high workload require allocation of respective resources.


Synovial-Based Tests Outperform Serum Markers to Rule Out Infection in Total Knee Arthroplasty and Total Hip Arthroplasty: A Systematic Review and Meta-Analysis.
Total knee arthroplasty (TKA) provides successful results in most patients. Periprosthetic joint infection (PJI) accounts for up to 25% of failed TKAs needing revision. In clinical practice, consensus in diagnostic strategy for excluding or diagnosing PJI is still lacking. In this systematic review and meta-analysis, we aim to provide a simplified data-driven diagnostic strategy for aseptic knee and hip revision surgeons to rule out PJI in the outpatient clinic phase. A literature search in EMBASE, MEDLINE, PubMed, and Cochrane was conducted. Studies involving the diagnosis of PJI in patients with failed TKAs and total hip arthroplasties needing revision were identified. Only studies using the Musculoskeletal Infection Society criteria were included. Quality was assessed using MINORS criteria. Meta-analysis was performed for each diagnostic test identified in the included studies. Pooled estimates of diagnostic accuracy measures were calculated using a bivariate model and plotted in summary receiver-operator characteristic curves. Positive and negative predictive values were calculated in a hypothetical sample of patients with a given disease prevalence. Twenty-four studies met the inclusion criteria, describing a total of 2974 patients. Quality scores ranged from 13 to 19. Meta-analysis could be performed on 7 unique diagnostic tests. Highest pooled sensitivity and specificity were demonstrated for α-defensin with values of 86% and 96.6%, respectively. α-defensin and white blood cell count in synovial fluid demonstrate highest negative predictive value values. We recommend, in a clinical setting with low-intermediate prevalence of PJI, performing arthrocentesis and joint fluid analysis using α-defensin and/or white blood cell count before revision TKA and revision total hip arthroplasty surgery to rule out PJI.


Is paroxysmal supraventricular tachycardia truly benign? Insightful association between PSVT and stroke from a National Inpatient Database Study.
Atrial fibrillation and flutter are well-known causes of stroke. Whether other atrial arrhythmias categorized as paroxysmal supraventricular tachycardia (PSVT) are associated with stroke is less clear. We aimed to evaluate the association of PSVT with ischemic and embolic stroke and its impact on short-term outcomes in hospitalized stroke patients. National Inpatient Sample database of the USA was used to assess the association of PSVT with ischemic stroke. Atrial fibrillation and flutter were excluded to minimize the confounding effects. The association of PSVT with stroke was evaluated using univariate and multivariate analysis. Subgroup analyses by gender, age, and stroke type were also performed. PSVT was associated with increased odds of overall ischemic stroke in univariate [OR 1.18 (95% CI 1.09-1.27) p < 0.001] analysis. No such association was observed in multivariate analysis (OR 1.06 (95% CI 0.98-1.14) p = 0.1) or with subgroup analysis by gender and age. However, PSVT was associated with embolic stroke in both univariate (OR 2.01 (95%CI 1.67-2.43, p < 0.001) and multivariate analysis (OR 1.7 (95%CI 1.4-2.14) p < 0.001) as well as in subgroup analyses by gender and age. Furthermore, the presence of PSVT was associated with increased mortality in embolic stroke (OR 4.11, CI 2.29 to 7.39, p < 0.001) and increased total hospital cost and length of hospital stay in all stroke types. PSVT is independently associated with higher prevalence of embolic stroke but not with overall ischemic stroke. Patients with embolic stroke in the presence of PSVT have worse in-hospital outcomes with increased mortality.


A general separation method of phenolic acids using pH-zone-refining counter-current chromatography and its application to oat bran.
pH-zone-refining counter-current chromatography technique for the separation of natural and synthetic mixtures has been widely used, especially for organic acids and alkaloids. Phenolic acids are very important compounds due to the potential treatment for a wide variety of diseases. However, there is not a general method for their separation. In this work, the conditions of pH-zone-refining counter-current chromatography, involving solvent systems, concentration of retainer and eluter, flow rate of mobile phase as well as sample pretreatment, were optimized to improve extraction efficiency and reduce separation time. Finally a general separation method for seven common phenolic acids has been established using pH-zone-refining counter-current chromatography. The separation of these phenolic acids was performed with a two-phase solvent system composed of methyl tert-butyl ether/acetonitrile/water at a volume ratio of 4.75: 0.25: 5, where 3mM trifluoroacetic acid was added to the organic stationary phase as a retainer and 3mM NH4OH was added to the aqueous mobile phase as an eluter. As a result, seven phenolic acids, including syringic acid, 4-hydroxyphenylacetic acid, vanillic acid, caffeic acid, p-hydroxybenzoic acid, ferulic acid and p-coumaric acid were successfully separated with the purities of 95.9%, 67.3%, 96.9%, 82.4%, 97.0%, 91.0%, and 97.2%, respectively. The established general method has been applied to the crude sample of oat bran pretreated with AB-8 resin. A total of 49.5mg of syringic acid, 109.2mg of p-coumaric acid and 184.5mg of ferulic acid were successfully purified in one run from 1.22g crude extract with the purities of 95.2%, 93.0%, and 91.8%, respectively.


Family Sex Trafficking Among the Bedia Caste of India: Defying the Dominant Human Trafficking Discourse.
Largely characterized as an urban issue, empirical studies of minor sex trafficking in rural communities-including India, a global hotspot for child sex trafficking-are exceptionally uncommon. Yet, the commercial sexual exploitation of children thrives in many rural Indian villages, fueled by caste discrimination, family tradition, and poverty. In response, this study aimed to investigate minor sex trafficking among a particular culturally unique and geographically isolated population, in relation to the dominant human trafficking literature. In-depth interviews were conducted with 31 female members of the Bedia caste-a unique population whose primary form of income is derived from participation in the rural sex trade. Framed by the social theory of intersectionality, we sought to (1) identify vulnerabilities for commercial sex industry entry among Bedia youth and (2) examine the cultural context of the commercial sex industry among the Bedia, with particular attention to the dominant victim/perpetrator paradigms. Implications for continued research, practice, and policy are included.


Foot-and-mouth disease virus causes a decrease in spleen dendritic cells and the early release of IFN-α in the plasma of mice. Differences between infectious and inactivated virus.
Foot-and-mouth disease (FMD) is a highly contagious and acute viral disease of cloven-hoofed animals. From an economical point of view, it is the most important disease of livestock worldwide. It is known that the virus interacts with dendritic cells, both in the natural host and in mice, but the impact of this interaction on the adaptive immune response is controversial. Currently available vaccines are based on inactivated forms of the FMD virus. Little is known about the differences between infectious and inactivated virus, in terms of dendritic cell subsets involved in immune response activation. The present work, which was carried out in the murine model, shows that live virus infection induces a reduction in splenic dendritic cell subsets. In addition, lymphocyte proliferation is inhibited in the early stages of infection associated with IFN-α induction, but is restored to normal values 5 days post-infection when pro-inflammatory cytokines was produced. In contrast, the inactivated virus increases the percentage of plasmacytoid dendritic cells in the spleen and the production of IL-10, which triggers the activation of a T regulatory response.


PGE1 and E3 show lower efficacies than E2 to β-catenin-mediated activity as biased ligands of EP4 prostanoid receptors.
The 2-series of prostaglandin E (PGE2 ) is regarded as a pro-cancer prostanoid, whereas the 1-series (PGE1 ) and the 3-series (PGE3 ) are considered to act as anti-cancer prostanoids. In the present study, we provide possible reasons why PGE1 and PGE3 , but not PGE2 , exert anti-cancer effects by focusing on each diverged E-type prostanoid (EP)4 receptor-mediated signaling pathway. PGE1 , PGE2 and PGE3 function as full agonists in terms of Gαs - and Gαi -protein-mediated signaling. However, PGE1 and PGE3 function as partial agonists of T-cell factor (TCF)/β-catenin (β-cat)-mediated activity, the well-known cancer-related signaling pathway. Furthermore, pretreatment with PGE1 or PGE3 almost completely reduces PGE2 -induced TCF/β-cat activity. These results provide a plausible reason why PGE1 and PGE3 function as anti-cancer prostanoids as a result of novel biased activity for EP4 receptors.


Recurrent, familial Reye-like syndrome with a new complex amino and organic aciduria.
Five of 13 siblings from a Jewish-Ashkenazi family suffered from recurrent Reye-like episodes. During attacks, these patients excreted alpha-keto-adipic, alpha-hydroxy-adipic, and alpha-aminoadipic acids, branched-chain keto acids and saccharopine in addition to lactic, pyruvic, and dicarboxylic acids characteristic of Reye syndrome. The serum concentrations of citrulline and alpha-amino-adipic acid were elevated and carnitine was at the upper limit of the normal range. Serum acetoacetate level was 4-5 times the beta-hydroxybutyrate level, but the pyruvate/lactate ratio was normal. Notably, plasma ketone bodies were lower than expected from the degree of catabolism. When the patients were symptom-free, no abnormal amino or organic acids in serum or urine were detected. These findings might be interpreted as a functional impairment at three different biochemical sites: fatty acid beta-oxidation, dehydrogenase complexes of the pyruvic, alpha-ketoglutaric, alpha-ketoadipic, and branched-chain keto acids, and pyruvate carboxylase. We suggest that in this hereditary disorder a toxic substance, exogenously or endogenously derived, interfered at multiple sites in different metabolic pathways.


Detection and isolation of MUC1 mucin from larynx squamous cell carcinoma.
The progression from uncontrolled cell proliferation to invasion and metastasis of epithelial tumors is partially understood. Alteration of epithelial mucin expression have been described in different malignant localizations but only few attempts have been made to identify mucin expression in malignant laryngeal tumors. In the present report, results are shown of studies on the expression of mucins and carbohydrate related antigens in laryngeal cancer and on the isolation of MUC1 mucin from this tumor tissue. Malignant laryngeal specimens were processed for immunohistochemical analysis and for extranuclear membrane fractions (ENM) which were obtained by ultracentrifugation. Subsequently, ENM samples were centrifuged in density-gradient; the analysis of fractions was performed by means of SDS-PAGE and Western-blotting. The panel of monoclonal antibodies (MAbs) included anti MUC1 mucin, anti Lewis x, anti sialyl Lewis x, anti Lewis y, anti MUC-5B, anti oral mucin (gp230), anti Tn hapten, anti p53 and anti cytokeratins. By immunohistochemistry, it was possible to detect MUC1 mucin, Lewis x and Lewis y showing strong reactions while sialyl-Lewis x and Tn antigen only reacted weakly in a few cells; cytokeratins were detected in all samples. In ENM derived fractions obtained by CsCl centrifugation, MUC1 was demonstrated by Western blotting. (1) laryngeal cancer antigenic expression comprises mostly MUC1 mucin, Lewis x, Lewis y as well as Tn antigen and (2) the methodology here employed is useful to isolate MUC1 from tumor samples.


Prosthetic repair of incisional hernia combined with elective bowel operation.
Incisional hernia repair with mesh is considered a clean operation and it is not recommended to be perfomed at the same time with a potentially contaminated operation. The aim of this study is to assess the short-term results of a group of patients who underwent a colon operation and simultaneous incisional hernia repair with an onlay polypropylene mesh technique. From Novemberto June 2006, 19 patients underwent incisional hernia repair with polypropylene mesh, with simultaneous colonic operation. In 13 patients reestablishment of bowel continuity after a Hartmann procedure was done, whereas in four patients a loop colostomy was closed. Two patients underwent colectomy for cancer. Post-operatively one patient had a seroma and two others had wound infections which required mesh removal. The mean follow-up was 70.15 +/- 48.40 months (range 3 to 142 months). During this period five patients died, four from progression of malignancy and one from myocardial infarction. Three patients (15.78%) developed recurrence, two patients with previous Hartmann's operation for complicated diverticulitis and wound infection and the third patient due to inappropriate mesh fixation with buttonhole hernia development. Prosthetic repair of incisional hernias can be safely performed simultaneously with a colonic operation, with an acceptable rate of infectious complications and recurrence. It is unjustifiable to avoid the use of mesh in a potentially contaminated field when an appropriate technique is used.


Identification of calcium-inducible genes in primary keratinocytes using suppression-subtractive hybridization.
Terminal differentiation in epidermal keratinocytes involves major biochemical changes including the expression of many new differentiation-specific genes. To further understand this process, we performed suppression-subtractive hybridization of keratinocytes cultured under high-calcium condition, known to induce differentiation in vitro. We randomly isolated 300 clones representing 90 different genes. By reverse Northern blot analyses, 20 different genes were found to be overexpressed, of which 13 were confirmed as differentially expressed genes during keratinocyte differentiation by Northern blot analysis. Of those, five genes, transglutaminase 1, keratin 6, interleukin-1 receptor antagonist, kallikrein 7, and heat shock protein 27, are known to be up-regulated during epidermal differentiation. Six genes, ferritin-L chain, ribosomal protein S6, tumor-associated calcium signal transducer 2, neuroendocrine secretory protein 55, phosphoserine aminotransferase, and neutrophil gelatinase-associated lipocalin, heretofore were not known to be up-regulated during keratinocyte differentiation. We also identified two novel genes. One of these maps to chromosome 1q21 of the epidermal differentiation complex, and its expression level was strongly increased in differentiating keratinocytes. These differentially expressed genes may provide significant opportunities for further understanding of the epidermal keratinocyte differentiation.


Transition-Metal-Free Cyclopropanation of 2-Aminoacrylates with N-Tosylhydrazones: A General Route to Cyclopropane α-Amino Acid with Contiguous Quaternary Carbon Centers.
Cyclopropanation of 2-aminoacrylates with N-tosylhydrazones could proceed smoothly under transition-metal-free conditions via a [3 + 2] cycloaddition process. This robust protocol exhibits excellent generality, delivering a wide spectrum of cyclopropane α-amino acid esters bearing contiguous quaternary carbon centers in high yields and diastereoselectivities. With these readily available products, the steric convergence of cyclopropane α-amino acids could be readily obtained.


A multi-layer microchip for high-throughput single-cell gene expression profiling.
Microfluidics or Bio-MEMS technology offers significant advantages for performing high-throughput screens and sensitive assays. The ability to correlate single-cell genetic information with cellular phenotypes is of great importance to biology and medicine because it holds the potential to gain insight into disease pathways that is unavailable from ensemble measurements. Previously, we reported two kinds of prototypes for integrated on-chip gene expression profiling at the single-cell level, and the throughput was designed to be 6. In this work, we present a five-layer microfluidic system for parallelized, rapid, quantitative analysis of RNA templates with low abundance at the single-cell level. The microchip contains two multiplexors and one partitioning valve group, and it leverages a matrix (6 × 8) of quantitative reverse transcription polymerase chain reaction (RT-qPCR) units formed by a set of parallel microchannels concurrently controlled by elastomeric pneumatic valves, thereby enabling parallelized handling and processing of biomolecules in a simplified operation procedure. A comprehensive metallic nanofilm with passivation layer is used to run polymerase chain reaction (PCR) temperature cycles. To demonstrate the utility of the approach, artificial synthesized RNA templates (XenoRNA) and mRNA templates from single cells are employed to perform the 48-readout RT-qPCRs. The PCR products are imaged on a fluorescence microscope using a hydrolysis probe/primer set (TaqMan). Fluorescent intensities of passive reference dye and a fluorescein amidite reporter dye are acquired and measured at the end of PCR cycles.


Geographic Variation in the Use of Diagnostic Testing of Patients with Newly Diagnosed Open-Angle Glaucoma.
To determine the extent of geographic variation in the proportion of patients with newly diagnosed open-angle glaucoma (OAG) undergoing visual field (VF) testing, fundus photography (FP), and other ocular imaging (OOI) among patients residing in different US communities. Retrospective, longitudinal cohort study. All enrollees with newly diagnosed OAG enrolled in a managed care network between 2001 and 2014. We identified all persons in the plan with incident OAG residing in 201 communities across the United States. All communities contributed ≥20 enrollees. The proportion of enrollees undergoing ≥1 VF test, FP, OOI, and no testing of any type in the 2 years after first OAG diagnosis was determined for each community, and comparisons were made to assess the extent of variation in use of diagnostic testing among patients residing in the different communities. Receipt of VF testing, FP, OOI, or none of these tests in the 2 years after initial OAG diagnosis. Of the 56675 enrollees with newly diagnosed OAG, the mean proportion of patients undergoing VF testing within 2 years of initial diagnosis was 74%±7%, ranging from as low as 51% in Rochester, Minnesota, to as high as 95% in Lancaster, Pennsylvania. The mean proportion undergoing OOI was 63%±10% and varied from 34% in Palm Springs/Rancho Mira, California, to 85% in Charleston, South Carolina. The mean proportion receiving FP was 26%±10% and ranged from as low as 3% in Fresno, California, to as high as 57% in Harlingen, Texas. The proportion undergoing no glaucoma testing ranged from 0% in Binghamton, New York, to as high as 35% in 2 other communities. In many US communities, a high proportion of patients are undergoing testing according to established practice guidelines. However, in several communities, less than 60% of patients with newly diagnosed OAG are undergoing VF testing in the 2 years after initial OAG diagnosis, and in a few communities >1 in 4 patients have no record of glaucoma diagnostic testing of any type. Additional research is needed to understand factors driving this variation in practice patterns and its impact on patient outcomes.


Anti-inflammatory activity of superoxide dismutases: inhibition of carrageenan induced edema in rats.
Eighteen different superoxide dismutases from procaryote, plant, fish, bird and mammalian species have been tested for anti-inflammatory activity in the rat paw pad carrageenan-induced inflammation model. Very large differences in activity are observed. Homologous rat Cu-SOD is not active and indeed shows slight pro-inflammatory activity. The different SODs have different iso-electric values, different metals (Cu, Mn or Fe) at the active centre, different molecular weights and different circulation lifetimes. Biological activity is a function of amino acid sequence rather than of such secondary parameters.


Orbital leiomyoma: a case report with clinical, radiological and pathological correlation.
Orbital leiomyoma is a benign tumour of smooth muscle origin that can be clinically, radiologically and histopathologically difficult to diagnose due to the rarity of leiomyomas in this location. A 74-year-old woman presented with a 2-month history of painless proptosis. Computed tomography and magnetic resonance imaging studies revealed a well-circumscribed intraconal lesion separate from the optic nerve, and not eroding adjacent bone. A right lateral orbitotomy via a lid crease incision was performed to remove the firm lobulated lesion. Postoperatively there was an obvious decrease in proptosis. Microscopically the lesion was a well-circumscribed tumour composed of spindle cells. There were no light microscopic features to suggest malignancy, and immunohistochemistry was used to confirm the diagnosis of leiomyoma. Leiomyoma is a tumour that is rarely found in the orbit most likely due to the paucity of smooth muscle in this location. The differential diagnosis is wide, and the role of radiology, histology and immunohistochemistry in the diagnosis of leiomyoma is discussed. Surgical excision in this case excluded malignancy, provided a definitive diagnosis of a rare entity and resulted in marked clinical improvement.


Intensity of treatment, end-of-life care, and mortality for older patients with severe traumatic brain injury.
The Eastern Association for the Surgery of Trauma (EAST) recommends that clinicians consider limiting further aggressive treatment in geriatric patients with severe traumatic brain injury (TBI) who do not improve in 72 hours (nonresponders) owing to their poor prognosis. However, little is known about how these guidelines are followed in practice. This study compared mortality and patient care among geriatric patients with severe TBI classified as "responders" and "nonresponders" 72 hours after injury. Retrospective review of patients 65 years or older at a Level I trauma center with severe TBI (GCS < 8) from 2011 to 2014. We compared in-hospital mortality, end-of-life (EOL) decision making, discharge functional status, and 12-month survival in responders (GCS > 8 at 72 hours) and nonresponders (GCS ≤ 8 at 72 hours). Of 90 patients, 29 (32%) died within 3 days of injury, 29 (32%) were nonresponders, and 32 (34%) were responders. An additional 19 patients (21%) died before hospital discharge, of whom 17 (89%) were nonresponders. Nonresponders had higher odds of in-hospital death (odds ratio, 31.8; 95% confidence interval [CI], 3.71-272.9; p = 0.002). Family meetings to discuss goals of care were more common in the nonresponder group (p < 0.001) and fewer nonresponders were full code at discharge or death (p < 0.001). There were no significant differences in functional status at discharge. Among patients discharged alive, there were no differences in 12-month survival. The responder/nonresponder dichotomy identifies patients with higher in-hospital mortality outcomes and is associated with differences in EOL decision making. However, functional impairment and poor survival were prevalent, irrespective of neurologic status at 72 hours. Prognostic/epidemiologic study, level III; therapeutic study, level IV.


Adeno associated viral vector-mediated expression of somatostatin in rat hippocampus suppresses seizure development.
Somatostatin (SST) has been suggested to play an important role in maintaining hippocampal homeostasis by modulating excitatory neurotransmission. The putative anticonvulsant role for SST was tested in an electrical amygdala kindling model. SST was cloned into serotype 5 of the adeno-associated viral (AAV) vector and delivered bilaterally into the hippocampus of adult male Sprague Dawley rats that were subsequently electrically kindled. Behavioral severity and duration of kindled seizures was compared to uninjected and GFP-injected control rats. Results demonstrated that 70% of SST treated animals did not experience class IV or V seizures without affecting the threshold for individual stimulation-evoked seizures. This result was significantly different from control groups where 100% of animals reached class V seizures. No difference in the number of stimulations required to reach the first class I-III seizures was observed in the experimental cohort relative to age-matched controls. These preclinical results suggest a putative role for SST as an anticonvulsant therapeutic modality for epilepsy.


Development of an instrument to assess social functioning in dementia: The Social Functioning in Dementia scale (SF-DEM).
Social functioning is a core domain in the life of people with dementia, but there is no accepted instrument to measure it. We aimed to develop the Social Functioning in Dementia (SF-DEM) scale and test its psychometric properties for assessing social function in people with dementia. We interviewed people with mild dementia and family caregivers to develop patient and caregiver-rated SF-DEM versions and refined them through interviews with health care professionals. We tested its psychometric properties in 30 dyads of people with dementia and family caregivers. Both SF-DEM versions had content validity and demonstrated concurrent validity against a single item rating overall social functioning (patient rated r = 0.42, 95% CI [0.07-0.68]; caregiver rated r = 0.59, 95% CI [0.29-0.78]). All participants found it acceptable. Analyses showed reliability (test-retest, inter-rater, internal consistency) and indications of responsiveness to change. SF-DEM shows promise as a valid, reliable, acceptable measure of social functioning in dementia.


A deep learning-based hybrid model of global terrestrial evaporation.
Terrestrial evaporation (E) is a key climatic variable that is controlled by a plethora of environmental factors. The constraints that modulate the evaporation from plant leaves (or transpiration, Et) are particularly complex, yet are often assumed to interact linearly in global models due to our limited knowledge based on local studies. Here, we train deep learning algorithms using eddy covariance and sap flow data together with satellite observations, aiming to model transpiration stress (St), i.e., the reduction of Et from its theoretical maximum. Then, we embed the new St formulation within a process-based model of E to yield a global hybrid E model. In this hybrid model, the St formulation is bidirectionally coupled to the host model at daily timescales. Comparisons against in situ data and satellite-based proxies demonstrate an enhanced ability to estimate St and E globally. The proposed framework may be extended to improve the estimation of E in Earth System Models and enhance our understanding of this crucial climatic variable.


Reactive oxygen species induce phosphorylation of serine 118 and 167 on estrogen receptor alpha.
Estrogen receptor alpha (ERalpha) is a well-known target for signaling pathways originating from growth factor receptors. Reactive oxygen species (ROS) can induce activation of extracellular response kinase 1/2 (Erk1/2) and protein kinase B (Akt). Both kinases have been implicated in the phosphorylation of serine 118 and serine 167 on ERalpha, respectively. This activity may lead either to ligand-independent activation of ERalpha or down-regulation of ERalpha and may contribute to development of the resistance to endocrine therapy. Treatment of MCF-7 human breast cancer cells with glucose oxidase (GO, 0.1 un/ml) induced transient phosphorylation of serine 118 and serine 167. The increase in expression of p-ser118-ERalpha was 355 +/- 98% (mean +/- SD) and of p-ser167-ERalpha was 632 +/- 355%. These effects were enhanced in Her2 over-expressing MCF7 cells. ERalpha expression declined to 63 +/- 20% within the first 90 min of treatment and was below 10% 24 h later. ROS induced phosphorylation of ERalpha resulted in decreased expression of pS2 and progesterone receptor. Activation of Erk1/2 and Akt was transient with highest levels of Erk1/2 being 595 +/- 143% and p-Akt 311 +/- 125%. Inhibition of Erk1/2 by U0126 (10 microM) decreased p-ser118-ERalpha by 51.7 +/- 8.5% and decreased p-ser167-ERalpha by 41.9 +/- 16.9% whereas inhibition of Akt by LY294002 (20 microM) and wortmannin (500 nM) or by siRNA knock-down, had no effect on p-ser167-ERalpha expression. Our data show for the first time that ROS can induce post-translational modifications of ERalpha at serine 118 and serine 167, and may lead to ERalpha down-regulation in human breast cancer cells. Both the phosphorylation and consequent down-regulation of ERalpha may be a mechanism associated with development of endocrine therapy resistance.


Voltage-gated potassium currents in myelinating Schwann cells in the mouse.
1. The whole-cell variation of the patch-clamp technique was used to record ionic currents in Schwann cells obtained from enzyme-treated mouse sciatic nerves before and after the onset of myelination. 2. Only outward currents were evoked in embryonic Schwann cells, which had no myelin, at membrane potentials more positive than -40 mV. Neonatal myelinating cells developed depolarization-activated outward currents and hyperpolarization-activated inward currents. For large hyperpolarizations below -160 mV, inward currents exhibited a sag following a peak which appeared to be mainly due to Na+ blockade. 3. Membrane potentials of neonatal myelinating cells were more negative than those of embryonic cells. The depolarization of the membrane potentials per 10-fold increase in external K+ concentrations in neonatal myelinating cells was 57 mV which fits the Nernst equation for a K+ electrode. 4. Quinine (0.5-2 mM) blocked the outward currents in embryonic cells and Ba2+ (2 mM) blocked both outward and inward currents in neonatal myelinating cells leaving quinine-sensitive outward currents of the embryonic type. External Cs+ (5 mM) blocked mainly inward currents and internal Cs+ blocked outward currents. 5. Developmental changes of these voltage-gated K+ currents in myelinating cells showed that Ba2(+)-sensitive K+ currents disappeared rapidly during the first week of life in association with the membrane potential becoming more positive. In contrast, quinine-sensitive outward K+ currents of the embryonic type disappeared slowly during the first 3-4 weeks after birth. 6. It is concluded that neonatal myelinating Schwann cells developed new voltage-gated K+ channels, which are Ba2(+)-sensitive and set a new membrane potential, in addition to the voltage-gated K+ channels of embryonic type. The Ba2(+)-sensitive K+ channels in myelinating cells were suggested to play an important role in siphoning K+ ions accumulated in periaxonal space during nerve activities.


LSC measurements of the half-life of 40K.
Liquid scintillation counting techniques combined with the CIEMAT/NIST method have been applied to measure the specific activity of natural potassium salts. Samples have been prepared with three different scintillators. The individual atomic composition as well as the density of the cocktails have been taken into account for the efficiency calculation. With the specific activity the half-life is calculated to be T1/2 = 1.248(3) x 10(9) a. The result is in reasonable agreement with other measurement results provided the same isotopic concentration of 40K is used.


Hydroxycut(®) (herbal weight loss supplement) induced hepatotoxicity: a case report and review of literature.
Use of supplement and alternative drugs continues to thrive and is becoming an increasing cause of concern since many of these substances may have unexpected or unexplained medical consequences. We present below the first reported case of hepatotoxicity from Hydroxycut(®) in Hawaii.


Dopamine induces an optimism bias in rats-Pharmacological proof for the translational validity of the ambiguous-cue interpretation test.
Recent findings have revealed that pharmacological enhancement of dopaminergic (DA) function by the administration of a dopaminergic precursor (dihydroxy-l-phenylalanine; l-DOPA) increases an optimism bias in humans. This effect is due to l-DOPA's impairment of the ability to update beliefs in response to undesirable information about the future. To test whether an 'optimistic' bias is also mediated by dopamine in animals, first, two groups of rats received either a dopaminergic precursor, l-DOPA, or a D2 receptor antagonist, haloperidol, and were subsequently tested using the ambiguous-cue interpretation (ACI) paradigm. To test whether similar effects might be observed when manipulating another neurotransmitter implicated in learning about reward and punishment, we administered the serotonin (5-HT) reuptake inhibitor escitalopram to a third group of animals and the selective and irreversible tryptophan hydroxylase inhibitor para-chlorophenylalanine (PCPA) to a fourth group. The results of our study demonstrated that prolonged (2 weeks), but not acute, l-DOPA administration induced optimistic bias in rats. Neither acute nor chronic treatment with the other tested compounds had significant effects on the cognitive judgment bias of rats. The convergence of these results with human studies suggests the translational validity of the ambiguous-cue interpretation paradigm in testing the effects of pharmacological manipulations on cognitive judgment bias (optimism/pessimism) in rats.


Changes in occupational class differences in leisure-time physical activity and the contribution of retirement.
Occupational class differences in leisure-time physical activity (LTPA) are well recognized. Less is known about how these differences develop as individuals age, and how retirement is associated with this change. We investigated how occupational class differences in LTPA change in a cohort over a 15-17 years follow-up. We further examined, how the transition into mandatory or disability retirement contributed to the change in LTPA levels and occupational class differences. We used the data from the Helsinki Health Study surveying the aging City of Helsinki employees. In all, 8773 individuals were included in the analyses. We evaluated LTPA levels using weekly metabolic equivalent task (MET) hours and used generalized linear mixed effect models (GLMM) to estimate the development of LTPA levels. Commuting was included in the LTPA measure. Occupational class differences in LTPA emerged and widened during the follow-up. The physical activity levels decreased in the lower occupational class and slightly increased in the higher occupational class, resulting in a difference of 4.3 MET-hours at the end of follow-up, accounting for 50 min of brisk walking per week. The occupational class differences emerged during transition into mandatory retirement and persisted after this. Transition into disability retirement temporarily widened the occupational class differences in LTPA levels, but the differences diminished during the follow-up. Research on interventions to counteract the declining LTPA is needed to discover ways to prevent the widening of occupational health disparities during aging. The transition into old-age retirement could be an optimal period for focusing these interventions.


CK2 Phosphorylation Is Required for Regulation of Syntaxin 1A Activity in Ca2+-Triggered Release in Neuroendocrine Cells.
The polybasic juxtamembrane region (5RK) of the plasma membrane neuronal SNARE, syntaxin1A (Syx), was previously shown by us to act as a fusion clamp in PC12 cells, as charge neutralization of 5RK promotes spontaneous and inhibits Ca2+-triggered release. Using a Syx-based FRET probe (CSYS), we demonstrated that 5RK is required for a depolarization-induced Ca+2-dependent opening (close-to-open transition; CDO) of Syx, which involves the vesicular SNARE synaptobrevin2 and occurs concomitantly with Ca2+-triggered release. Here, we investigated the mechanism underlying the CDO requirement for 5RK and identified phosphorylation of Syx at Ser-14 (S14) by casein kinase 2 (CK2) as a crucial molecular determinant. Thus, following biochemical verification that both endogenous Syx and CSYS are constitutively S14 phosphorylated in PC12 cells, dynamic FRET analysis of phospho-null and phospho-mimetic mutants of CSYS and the use of a CK2 inhibitor revealed that the S14 phosphorylation confers the CDO requirement for 5RK. In accord, amperometric analysis of catecholamine release revealed that the phospho-null mutant does not support Ca2+-triggered release. These results identify a functionally important CK2 phosphorylation of Syx that is required for the 5RK-regulation of CDO and for concomitant Ca2+-triggered release. Further, also spontaneous release, conferred by charge neutralization of 5RK, was abolished in the phospho-null mutant.


Species richness and the analytic geometry of latitudinal and altitudinal gradients.
Extensive empirical work has shown that species richness decreases roughly exponentially or quadratically with latitude. What appears to be a latitudinal gradient in fact may simply be a negative correlation of latitude with area at that latitude, due to convergence of lines of meridian at the poles. There is simply less area at high latitudes, which means fewer niches and fewer opportunities for speciation, hence diminished biodiversity at high latitudes. Similarly, analytic geometry of a cone shows that species number should decrease linearly with altitude on a conical mountain. Here, I provide an explicit mathematical model of the area hypothesis of species richness along latitude and altitude gradients. I re-analyze a previously published latitudinal gradient dataset and show that species number is a linear function of the predicted area and that species number is more fully explained by predicted area than by a quadratic function of latitude. However, analytic geometry is not needed if precise measures of area are known.


Incidence and severity of experimental allergic encephalomyelitis and cerebral prostaglandin synthesis in essential fatty acid deficient and aspirin-treated rats.
Experimental allergic encephalomyelitis (EAE) was induced in rats of the Lewis strain fed diets adequate or deficient in essential fatty acids (EFA). After induction of the disease, the diets were supplemented with aspirin (3.75 g/kg diet), and the effects of the drug on the course of EAE and on the synthesis of prostaglandin F (PGF) by brain slices from diseased animals and their Freund controls were examined. Aspirin supplementation delayed the onset of EAE in both dietary groups. EFA-deficient rats experienced an incidence and severity of the disease similar to that of aspirin-free, EFA-deficiet rats, while the EFA-adequate group showed a greater severity but not an increased incidence, compared to aspirin-free controls. Aspirin treatment led to an increased PGF production by brain slices from rats on either diet and not subjected to an immunochallenge. When the diet was deficient in EFA, challenge with antigen plus adjuvant or adjuvant alone tended to decrease PGF synthesis by brain slices, and when the diet was adequate in EFA, immunochallenge caused a marked depression on PGF synthesis. It was concluded that the PG synthetase inhibitor aspirin can alter the course of EAE in the rat, providing further evidence that PGs or related metabolites may be involved in the immune response in this disease.


Cloning of a new member of the peroxisome proliferator-activated receptor gene family from mouse liver.
Peroxisome proliferators are postulated to elicit predictable pleiotropic responses in the liver by activating a peroxisome proliferator-activated receptor (PPAR). PPARs from mouse liver (mPPAR), rat liver (rPPAR), and Xenopus liver (xPPAR gamma) have been cloned recently. We now report the cloning of a new member from mouse liver which we designate mPPAR gamma. mPPAR gamma cDNA contained an open reading frame encoding a 475-amino acid protein exhibiting 75% amino acid similarity to xPPAR gamma, while it showed only 55% identity with mPPAR. The ligand-binding and DNA-binding domains are best conserved between mPPAR gamma, mPPAR, and xPPAR gamma. Like rPPAR, mPPAR gamma is able to impart peroxisome proliferator responsiveness to the promoter of peroxisomal bifunctional gene, which encodes the second enzyme of the peroxisomal fatty acid beta-oxidation system. Northern blot analysis revealed high expression of mPPAR gamma gene in mouse liver, kidney, and heart and low expression in the lung, testis, brain, skeletal muscle, and spleen. In mice treated with ciprofibrate, a peroxisome proliferator, a 2-fold increase in mPPAR gamma mRNA was observed in the liver and kidney. The presence of two PPARs in the mouse liver suggests the possibility of multiple signaling pathways for the peroxisome proliferator-induced pleiotropic responses.


Proteomics of sea bass skin-scales exposed to the emerging pollutant fluoxetine compared to estradiol.
Teleost fish skin-scales are essential for protection and homeostasis and the largest tissue in direct contact with the environment, but their potential as early indicators of pollutant exposure are hampered by limited knowledge about this model. This study evaluated multi-level impacts of in vivo exposure of European sea bass to fluoxetine (FLX, a selective serotonin-reuptake inhibitor and an emerging pollutant) and 17β-estradiol (E2, a natural hormone and representative of diverse estrogenic endocrine-disrupting pollutants). Exposed fish had significantly increased circulating levels of FLX and its active metabolite nor-FLX that, in contrast to E2, did not have estrogenic effects on most fish plasma and scale indicators. Quantitative proteomics using SWATH-MS identified 985 proteins in the scale total proteome. 213 proteins were significantly modified 5 days after exposure to E2 or FLX and 31 were common to both treatments and responded in the same way. Common biological processes significantly affected by both treatments were protein turnover and cytoskeleton reorganization. E2 specifically up-regulated proteins related to protein production and degradation and down-regulated the cytoskeleton/extracellular matrix and innate immune proteins. FLX caused both up- and down-regulation of protein synthesis and energy metabolism. Multiple estrogen and serotonin receptor and transporter transcripts were altered in sea bass scales after E2 and/or FLX exposure, revealing complex disruptive effects in estrogen/serotonin responsiveness, which may account for the partially overlapping effects of E2 and FLX on the proteome. A large number (103) of FLX-specifically regulated proteins indicated numerous actions independent of estrogen signalling. This study provides the first quantitative proteome of the fish skin-scale barrier, elucidates routes of action and biochemical and molecular signatures of E2 or FLX-exposure and identifies potential physiological consequences and candidate biomarkers of pollutant exposure, for monitoring and risk assessment.


Purification of cytosolic glutathione transferases from Schistocephalus solidus (plerocercoid): interaction with anthelmintics and products of lipid peroxidation.
Glutathione (GSH) transferase isoenzymes have been partially resolved from the cytosol of Schistocephalus solidus (plerocercoid) by GSH affinity chromatography and chromatofocusing at pH 7-5. The presence of isomeric forms was also suggested by analytical isoelectric focusing and high-performance liquid chromatography (HPLC). Gel filtration and sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that GSH transferase forms were dimers with a subunit size of approximately 24 kDa. The major GSH transferase form in S. solidus (plerocercoid) showed greater biochemical relationship to the Mu family of mammalian GSH transferase compared to the mammalian Alpha or Pi families. The major subunit purified by GSH affinity chromatography and reversed-phase HPLC also showed high N-terminal homology with the Mu family. A minor GSH transferase form appeared more biochemically related to the Alpha family with respect to substrate specificity and inhibitor sensitivity. The major GSH transferase was inhibited by haematin-related compounds, bile acids and a number of anthelmintics including members of the benzimidazole and phenol-based class of compounds. The major GSH transferase had conjugating activity with members of the trans, trans-2,4-alkadienal and trans-2-alkenal series, secondary products of lipid peroxidation.


The needs of carers: a comparison between eating disorders and schizophrenia.
This pilot study compares the level of distress and the need for support between carers of patients with anorexia (AN) and bulimia nervosa (BN) and carers of patients with schizophrenia. Thirty-two carers of patients suffering from AN and BN and 30 carers of patients with schizophrenia filled out the general health questionnaire (GHQ-12) and the burden inventory (BI). In addition, they were interviewed with a semi-structured research interview, the carers' needs assessment (CNA), to assess relevant problem areas as well as the need for helpful interventions. Patients with an eating disorder were interviewed with the eating disorder examination (EDE) and patients with schizophrenia with the positive and negative syndrome scale (PANSS) to assess the severity of the disorder. The mean duration of illness was 5.3 years in the patients with eating disorders and 7.3 years in the patients with schizophrenia. Most of the carers were mothers or partners. In the CNA we found high numbers of problems as well as high numbers of unmet needs for interventions. The most frequently mentioned problem areas in all groups of carers were "disappointment caused by the chronic course of the illness, concerns about the patient's future" and "problems in communication with the patient". The most frequently reported need for support in all groups was "counseling and support by a professional". Carers of patients with BN reported a significantly lower number of problems and of needs for interventions compared to the carers of patients with AN and schizophrenia. Also in the BI and the GHQ-12 we found significantly lower total scores in carers of patients with BN. Carers of patients with AN and schizophrenia did not differ significantly in any of the assessments. The severity or duration of illness of the patients did not correlate with the carers' burden or needs. Carers of patients with an eating disorder and carers of patients with schizophrenia are burdened with similar problems and have high levels of unmet needs which are usually not addressed in clinical practice. Carers of anorectic patients have higher levels of difficulties in various areas compared to carers of bulimic patients and did not differ from carers of patients with schizophrenia.


Sanitary factors and mycotoxin contamination in the argentinian wheat crop 1993/94.
In Argentina, due to climatic conditions, Fusarium head blight (FHB) caused by Fusarium graminearum, affected the 1993/94 wheat crop. To evaluate the severity of this disease, samples of wheat where gathered from four zones of the wheat area. Sanitary conditions and mycotoxin contamination were determined. One zone (IIN) was intensely affected by FHB with 90% of samples in grade III (bad quality). No samples were grade I (good quality). The other zones were less affected falling into grade I or II (moderate quality). In all samples tested F. graminearum was the most prevalent species singly or in combination with others. Zone II N, with a DON mean level of I1.26 ppm, did not fulfil aceptability limits, whereas zones IIS, III and IV with overall means of 2.12, 1.57 and 1.0 ppm, respectively, did. Statistical analysis showed a close relation between percentage FHB and DON contamination (r:-0.71, p<0.01) in infected samples.


Predominance of locally-produced prostaglandin E2 over prostacyclin in skin incisions in man.
Blood was collected from skin incisions made by the 'Simplate' technique in 8 healthy men. Prostaglandin (PG) E2, but not 6-oxo-PGF1 alpha, the stable hydrolysis product of prostacyclin (PGI2), was tentatively identified using capillary column gas chromatography/electron capture mass spectrometry. It was not possible to quantify PGE2 because of the small volumes of blood generated by this method. Larger blood samples were then collected from 22 skin incisions in 13 patients undergoing cardiothoracic surgery. Concentrations of PGE2 were substantially greater than 6-oxo-PGF1 alpha in every sample. 13,14-Dihydro-15-oxo-PGF2 alpha, a pulmonary metabolite of PGE2, was not elevated, indicating that the PGE2 was synthesised locally at the site of incision. In 6 further patients undergoing cardiac surgery, blood sampled from an antecubital vein before and during operation contained little or no PGE2. We conclude that a substantial proportion of the PGE2 in blood emerging from skin incisions may be formed locally by the traumatised microvessels, consistent with the hypothesis that PGE2 is the principle prostaglandin synthesised by human cutaneous microvessels in vivo.


Expression, purification, and characterization of phospholipase B1 from Candida albicans in Escherichia coli.
Candida albicans is an important fungal pathogen that causes a wide variety of human infections, ranging from mucocutaneous infections to life-threatening systemic infections. Phospholipase B1 (PLB1) has been reported to be directly responsible for C. albicans pathogenicity and is likely to be involved in the early steps of host invasion. Therefore, PLB1 could be a potential marker for diagnosis of C. albicans infection. In this study, PLB1 was expressed using an Escherichia coli expression system. Recombinant PLB1 is found in inclusion bodies and constitutes up to 38.4% of total insoluble protein. After refolding in a GSH/GSSG redox system, GST-tagged PLB1 was purified by GST-sepharose 4B affinity chromatography and then cleaved with thrombin to remove the GST-tag. The recombinant PLB1 was further purified by anion-exchange chromatography and reverse phase HPLC. The final yield of purified PLB1 was approximately 15.6 mg from 100 mL of bacterial cell culture, and its concentration was 784 μg/μL. The recombinant PLB1 could form a white precipitation zone on egg yolk agar plate, suggesting its phospholipase activity. Moreover, the maximum activity of PLB1 was 68 IU/mg at pH 6.0, 37 °C. Therefore, recombinant PLB1 has potential application in structural analytical studies, or diagnosis of C. albicans infection.


Effect of Tussilago farfara L. Polysaccharides on the Content of Hematopoietic Stem Cells (CD117+34+) in the Bone Marrow of Mice Treated with Cyclophosphamide.
Myelotoxicity is a serious side effect of anticancer drugs. The search for drugs that can reduce the hematological complications of chemotherapy through modulation of hematopoietic stem cells is an urgent task of oncopharmacology. In the present study we showed that administration of Tussilago farfara L. polysaccharides to C57BL/6 mice treated with cyclophosphamide can increase the number of hematopoietic stem cells (CD117+34+) in the bone marrow.


Dysfunction of murine dendritic cells induced by incubation with tumor cells.
In vivo studies showed that dendritic cell (DC) dysfunction occurred in tumor microenvironment. As tumors were composed of many kinds of cells, the direct effects of tumor cells on immature DCs (imDCs) are needed for further studies in vitro. In the present study, bone marrow-derived imDCs were incubated with lymphoma, hepatoma and menaloma cells in vitro and surface molecules in imDCs were determined by flow cytometry. Then, imDCs incubated with tumor cells or control imDCs were further pulsed with tumor lysates and then incubated with splenocytes to perform mixed lymphocyte reaction. The DC-dependent tumor antigen-specific T cell proliferation, and IL-12 secretion were determined by flow cytometry, and enzyme-linked immunosorbent assay respectively. Finally, the DC-dependent tumor-associated antigen-specific CTL was determined by enzyme-linked immunospot assay. The results showed that tumor cell-DC incubation down-regulated the surface molecules in imDCs, such as CD80, CD54, CD11b, CD11a and MHC class II molecules. The abilities of DC-dependent antigen-specific T cell proliferation and IL-12 secretion were also decreased by tumor cell incubation in vitro. Most importantly, the ability for antigenic-specific CTL priming of DCs was also decreased by incubation with tumor cells. In the present in vitro study demonstrated that the defective abilities of DCs induced by tumor cell co-incubation and the co-incubation system might be useful for future study of tumor-immune cells direct interaction and for drug screen of immune-modulation.


Elemental profile of dietary supplements and agricultural byproducts evaluated by neutron activation analysis.
Dietary supplements and agricultural byproducts were characterized by neutron activation analysis. The nutritional potential of supplements was evaluated according to alternative and commercial categories, using analysis of variance and cluster analysis, and recommended dietary intake for children. The results indicated statistically significant differences between both categories for the elements Cs, K, Na, and Rb. For the nutritional elements Ca, Co, Fe, K, Na, and Zn, the categories were similar in cluster analysis. The similarity between elemental profiles of alternative supplements and agricultural byproducts was calculated using a dissimilarity matrix, showing that rice and wheat are the predominant ingredients.


Antiarrhythmic evaluation of naloxone against acute coronary occlusion-induced arrhythmias in pigs.
Naloxone pretreatment (1.0, 3.0 and 10.0 mg/kg i.v.) failed to protect anesthetized pigs from cardiac arrhythmias including ventricular fibrillation (VF) and death following acute occlusion (20 min) or reperfusion of the left anterior descending coronary artery. These findings suggest that opiate-like substances possibly released by the ischemic myocardium do not contribute significantly to the etiology of cardiac arrhythmias, or sudden death associated with the early stages of myocardial infarction in pigs. The effectiveness of naloxone in preventing acute ischemia-induced arrhythmias in rats may be due to mechanisms other than opiate-receptor blockade.


Aberrant Myeloid Differentiation Contributes to the Development of Osteoporosis in Neurofibromatosis Type 1.
Neurofibromatosis type 1 (NF1), also known as von Recklinghausen disease, is a common autosomal dominant genetic disorder affecting approximately 1 in 3000 individuals worldwide. NF1 results from heritable or spontaneous mutations of the NF1 tumor suppressor gene. NF1 encodes the protein neurofibromin, which functions to negatively regulate Ras-activity. Approximately 50 % of NF1 patients develop osteopenia or osteoporosis, resulting in significantly increased rates of long-bone fracture and morbidity. While defective osteoblast bone anabolism has been implicated as a central factor in the pathogenesis of NF1 associated skeletal deficits, recent data suggest that NF1 (Nf1) haploinsufficiency within the hematopoietic compartment, particularly in osteoclasts and myeloid progenitors, plays a pivotal role in engendering NF1 osseous manifestations. In this chapter, we review the latest data from clinical studies and murine models delineating a critical role for hematopoietic compartment, myeloid progenitors of NF1 (Nf1) haploinsufficient and their progeny-osteoclasts, in the pathogenesis of NF1 associated osteopenia/osteoporosis and discuss putative targets for future therapeutics.


The ontogeny of sugar transport in kidney.
Concentrative uptake of alpha-methyl-d-glucoside (AMG) by isolated renal tubule fragments from the newborn Sprague-Dawley rat has been demonstrated and the validity of this phenomenon confirmed by an in vivo demonstration of AMG uptake by the newborn kidney cortex. A kinetic analysis of the entry phenomenon in the newborn tubule reveals the presence of two distinct membrame transport systems for AMG, only one of which is present in the adult tubule. All transport of sugar in both newborn and adult tubules was phlorizin-sensitive, but was only partially ingibited in Na+-free buffer. Glucose was shown to inhibit uptake competitively on the shared, high capacity system. Uptake on the high affinity system in the newborn represents 15-20% of the total at physiologic sugar concentrations. It is concluded that active sugar transport is a characteristic of the newborn rat kidney and that the isolated tubule preparation is a more accurate reflection of this phenomenon than is the renal cortical slice.


An upper limit to the abundance of aquatic organisms.
The maximum density achievable by aquatic organisms is an inverse linear function of their body size. As a consequence, the maximum achievable biomass is independent of body size, and is 2 orders of magnitude higher than the biomass in natural populations. The minimum interorganismic terorganismic distance, calculated from the maximum density to allow comparison between aquatic and terrestrial organisms, scales as the 1/3 power of body size in both habitats. The similarities in the interorganismic distance of terrestrial and aquatic plant and animal communities suggest a fundamental regularity in the way organisms use the space.


Food intake at age 8. 2. Frequency, company and place of meals.
In a sample of 141 healthy 8 year old children, over 95% had three main meals on each of the 4 days of the food record. Two-thirds had an afternoon snack and nearly one-half a morning snack on each of the 4 days, but only one-quarter had supper on each day. Seventy-one per cent of all meals were eaten at home with the family, 20% at school with friends, and less than 1% of meals were eaten at a restaurant or fast food outlet. These data show that amongst children of primary school age, take away sources of food make a very small contribution to the diet, and that eating habits at home are of greatest importance to the child's diet.


Differences in genetic population structures of Plasmodium falciparum isolates from patients along Thai-Myanmar border with severe or uncomplicated malaria.
There have been many reports on the population genetic structures of Plasmodium falciparum from different endemic regions, but few studies have examined the characteristics of isolates from patients with different clinical outcomes. The population genetic structures of P. falciparum isolates from patients with either severe or uncomplicated malaria were examined. Twelve microsatellite DNA loci from P. falciparum were used to assess the population genetic structures of 50 isolates (i.e., 25 isolates from patients with severe malaria and 25 from patients with uncomplicated malaria) collected in the Thai-Myanmar border area between 2002 and 2005. Genetic diversity and effective population sizes were greater in the uncomplicated malaria group than in the severe malaria group. Evidence of genetic bottlenecks was not observed in either group. Strong linkage disequilibrium was observed in the uncomplicated malaria group. The groups demonstrated significant genetic differentiation (P < 0.05), and allele frequencies for 3 of the 12 microsatellite loci differed significantly between the two groups. These findings suggest that the genetic structure of P. falciparum populations in patients with severe malaria differs from that in patients with uncomplicated malaria. The microsatellite loci used in this study were presumably unrelated to antigenic features of the parasites, but, these findings suggest that some loci may influence the clinical outcome of malaria.


Morphological and histochemical observations on the oocapt and oviducal transport of oocytes in paramphistomum cervi (Zeder, 1790) (Digenea: Trematoda).
The oocapt of Paramphistomum cervi has a narrow constriction followed by a wide lumen with a thickened wall and is surrounded by several unicellular flask-shaped gland cells opening into the lumen. The oocapt gland cells are filled with granules, the size and number of which vary in the body cells and its duct. Histochemical studies have shown that the oocapt gland cells secrete mainly mucoproteins but a few lipoprotein and phospholipid granules have also been observed. The passage of the oocyte through the lumen of the oocapt, appears to stimulate the oocapt gland cells to release their secretion which seems to be involved in transport of the oocyte through the oviduct.


Central gray and medial hypothalamic stimulation: correlation between escape behavior and unit activity.
A chronic experiment and an acute one were carried out in the same rat in order to get information on the neuronal events correlated with the escape latency (EL) induced by an electrical stimulation in the mesencephalic central gray (CG) or in the medial hypothalamus (MH). Escape latencies as well as unitary MH and CG neuronal responses (104 units in 25 rats) were studied as functions of the intensity and the pulse duration of a 50 pulses/sec stimulation train. The CG and/or the MH neuronal firing rate was either synchronized or diffusely altered, either inhibited or activated, by the CG or the MH stimulations. Many neuronal firing rate alterations were highly correlated with the escape speed (ES = 1/EL) induced by the same stimulation. Intensity duration trade-off functions were computed from both the EL and the unit recordings. Chronaxie determinations were performed from these data: the behaviorally determined chronaxies did not differ from the unitary ones (limit values: 0.05-0.42 msec). The discussion bear on the possible role of the observed neuronal alterations induced by CG or MH stimulations in relation to their behavioral effects.


A Method for Examining Geographic Variation When Denominators Are Unknown, Populations Vary, and Time is of the Essence: Back Surgery as an Example.
: Level of Evidence: 3.


Investigation on formulation and preparation of adenovirus encoding human endostatin lyophilized powders.
A recombinant adenovirus encoding human endostatin gene, E10A, has finished phase II trials for head and neck cancer. However, the rigid storage temperature (-80°C) and the toxicity of glycerol in the E10A liquid preparation limited its clinical application. In this study, lyophilization was applied to develop a stable E10A lyophilized powder without glycerol that is able to maintain biological activity at 4°C and suitable for intravenous administration. The E10A lyophilized formulations composed of nontoxic and already clinically used excipients were characterized in terms of the pH change during freezing, the eutectic melting temperature (T(eu)) and the collapse temperature (T(c)). Freeze thawing tests were carried out to examine the protective effect of various excipients during freezing. Mannitol and its combinations with sucrose or inulin showed effective protection of E10A. The E10A lyophilized powders were analyzed by particle size measurement, residual humidity quantification, infectivity assay and gene expression level. An optimized formulation (formulation I1) yielded a good recovery of 76% of the starting infectivity after lyophilization and 89% of the original infectivity after storage at 4°C for 180 days. Also the gene expression capability of E10A in formulation I1 was maintained after lyophilization. In addition, it was found that the matrix of amorphous excipients, mannitol combinations with sucrose or inulin, was indispensible in protecting E10A against the stress of freezing and dehydration. Hereby, the E10A lyophilized powder with eliminated glycerol toxicity and improved stability could enhance the applicability of E10A for cancer gene therapy through intravenous administration.


"To me, it's my life": medical communication, trust, and activism in cyberspace.
This paper studies the conversations and activities of an online support group for breast cancer sufferers and survivors and their supporters. Using communications medium theory and social capital theory, it examines the mundane and profound exchanges, the poignant self-disclosures, the creative expressions of solidarity, and the minor but not-insignificant political actions of people--initially strangers--who come together as a 'virtuous circle,' not only to assist with medical issues but also to meet emotional and even material needs. Sponsored by the Canadian nonprofit organization Breast Cancer Action Nova Scotia (BCANS), this virtual community has logged over a half million messages since 1996. Not every BCANS participant is an activist--many are just trying to grapple with their disease--but some find ways to shatter the professional "information monopoly," and to press for healthcare improvements. The study illustrates the scope, passion, and complexity of peer-to-peer medical communication in a virtual environment that promotes "thick trust". BCANS participants discuss with candor, warmth and even humor such painful topics as death and dying and the crises in intimate relationships brought about by a terminal illness. The sharing of confidences and fears enables participants to pool their 'collective intelligence' about many things, from how to cope with swelling, to how to think about end-of-life issues, to how to improve social policy.


Impaired processing of prosodic and musical patterns after right hemisphere damage.
The distinction between the processing of musical information and segmental speech information (i.e., consonants and vowels) has been much explored. In contrast, the relationship between the processing of music and prosodic speech information (e.g., intonation) has been largely ignored. We report an assessment of prosodic perception for an amateur musician, KB, who became amusic following a right-hemisphere stroke. Relative to matched controls, KB's segmental speech perception was preserved. However, KB was unable to discriminate pitch or rhythm patterns in linguistic or musical stimuli. He was also impaired on prosodic perception tasks (e.g., discriminating statements from questions). Results are discussed in terms of common neural mechanisms that may underlie the processing of some aspects of both music and speech prosody.


Bioinformatics identification of key candidate genes and pathways associated with systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by autoantibody production and multi-system involvement, but the etiology is largely unclear. This study aimed to elucidate candidate genes and pathways involved in SLE. Three original datasets GSE72509, GSE20864, and GSE39088 were downloaded from Gene Expression Omnibus (GEO) and the data were further integrated and analyzed. Subsequently, differentially expressed genes (DEGs) between SLE patients and healthy people were identified. And then we performed gene ontology (GO) function and pathway enrichment analyses of common DEGs, and constructed a protein-protein interaction (PPI) network with STRING database. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was carried out to validate the expression levels of candidate genes in blood samples from SLE patients and healthy controls. In total, 321 common DEGs were identified in SLE patients compared with healthy controls, including 231 upregulated and 90 downregulated genes. GO function analysis revealed that 321 common DEGs were mainly enriched in innate immune response, defense response, cytokine-mediated signaling pathway, response to interferon-alpha, and I-kappaB kinase/NF-kappaB signaling. Additionally, pathway enrichment analysis indicated that DEGs were mainly enriched in several signaling pathways associated with immune system and apoptosis, including RIG-I-like receptor signaling pathway, antigen processing and presentation, and p53 signaling pathway. The expression levels of candidate genes RPL26L1, FBXW11, FOXO1, and SMAD7 were validated by RT-qPCR analysis. The four hub genes including RPL26L1, FBXW11, FOXO1, and SMAD7 may play key roles in the pathogenesis and development of SLE. RIG-I-like receptor signaling pathway, antigen processing and presentation pathway, and p53 signaling pathway may be closely implicated in SLE pathogenesis. Collectively, these results may provide valuable novel markers or targets for the diagnosis and treatment of SLE.Key Points• Integrated bioinformatics analysis of three profile datasets based on SLE patients and healthy controls was performed and 321 common DEGs were identified.• The 321 common DEGs were mainly enriched in biological processes related to immune responses and inflammatory responses, including innate immune response, defense response, cytokine-mediated signaling pathway, response to interferon-alpha, I-kappaB kinase/NF-kappaB signaling, whereas the three most significant cellular components were oxidoreductase complex, AIM2 inflammasome complex, and ubiquitin ligase complex.• KEGG pathway enrichment analysis indicated that common DEGs were mainly enriched in several signaling pathways associated with immune system and apoptosis, including RIG-I-like receptor signaling pathway, antigen processing and presentation, and p53 signaling pathway.• Candidate genes RPL26L1, FBXW11, FOXO1, and SMAD7 may be closely involved in the pathogenesis and development of SLE and may provide valuable novel markers or targets for the diagnosis and treatment of SLE.


Dialuric acid autoxidation. Effects of transition metals on the reaction rate and on the generation of "active oxygen" species.
The autoxidation of dialuric acid, a process which is believed to be of crucial importance in the diabetogenic action of alloxan, was found to be strongly catalysed by copper, iron and manganese. Superoxide radical and hydrogen peroxide were generated in both the uncatalysed and the metal-catalysed reactions. In contrast, hydroxyl radical was formed during dialuric acid autoxidation only in the presence of added iron salts. Production of the latter radical was strongly inhibited by catalase but only weakly by superoxide dismutase, implying that the metal-catalysed Haber-Weiss reaction is of comparatively little importance in hydroxyl radical generation from dialuric acid.


Advances in Conifer Somatic Embryogenesis Since Year 2000.
This review compiles research results published over the last 14 years on conifer somatic embryogenesis (SE). Emphasis is placed on the newest findings that affect the response of seed embryos (typical explants) and shoot primordia (rare explants) to the induction of SE and long-term culture of early somatic embryos. Much research in recent years has focused on maturation of somatic embryos, with respect to both yield and quality, as an important stage for the production of a large number of vigorous somatic seedlings. Attempts to scale up somatic embryo production numbers and handling have resulted in a few bioreactor designs, the utility of which may prove beneficial for an industrial application. A few simplified cryopreservation methods for embryonal masses (EM) were developed as a means to ensure cost-efficient long-term storage of genotypes during clonal field testing. Finally, recent long-term studies on the growth of somatic trees in the field, including seed production yield and comparison of seed parameters produced by somatic versus seed-derived trees, are described.


Losing Thomas & Ella: A Father's Story (A Research Comic).
"Losing Thomas & Ella" presents a research comic about one father's perinatal loss of twins. The comic recounts Paul's experience of the hospital and the babies' deaths, and it details the complex grieving process afterward, including themes of anger, distance, relationship stress, self-blame, religious challenges, and resignation. A methodological appendix explains the process of constructing the comic and provides a rationale for the use of comics-based research for illness, death, and grief among practitioners, policy makers, and the bereaved.


Addressing students' emotional needs during the COVID-19 pandemic: a perspective on text versus video feedback in online environments.
This paper reflects on the findings of Borup et al. (Educ Technol Res Dev 63:161-184, 2015) regarding the efficiency and affect of text and video feedback in the context of the rapid shift to online education due to the COVID-19 pandemic. Based on reports of diminished mental wellness, increased depression, and anxiety among learners and instructors, this paper offers ideas on how to apply the findings from Borup et al. (Educ Technol Res Dev 63:161-184, 2015) from a combination of practice, research, design, and inclusion perspectives to ensure emotional support, mental wellness, and social presence during times of crisis, even at the expense of efficiency of instruction.


High concentrations of dissolved organic nitrogen and N-nitrosodimethylamine precursors in effluent from biological nutrient removal process with low dissolved oxygen conditions.
The presence of dissolved organic nitrogen (DON) in biological nutrient removal (BNR) effluent has led to increased concern about its adverse effects on wastewater discharge and reuse applications. Previous studies have demonstrated efficient biological inorganic nitrogen removal in BNR under low dissolved oxygen (DO) conditions; however, information on DON is scarce. This study investigated low-DO effects on DON and N-nitrosodimethylamine (NDMA) precursor concentrations in BNR effluents. Identical BNR reactors consisting of an external real-time DO intelligent control system were operated at three different DO concentrations (0.3, 1.0, and 4.0 mgO2/L). Surprisingly, significantly higher values of effluent DON (p<0.05, t-test) and NDMA precursors (p<0.01, t-test) were observed at lower DO levels. Ultrahigh-resolution mass spectrometry analysis showed that molecules produced by microbes at low-DO levels exhibited high proteins/amino sugars-like and low normal oxidation state of carbon characteristics, which possibly acted critical roles in NDMA formation. Furthermore, path analysis by partial least-squares path modeling suggested that NDMA formation potential had strong associations with microbe-DON network stability of microbe-DON co-occurrence interactions (r=0.979, p<0.01). These results highlight the necessity of reconsidering the feasibility of BNR systems operating at low-DO concentrations considering the adverse effects of DON on wastewater discharge and reuse applications.


[PCPT trial: expected bias and their management].
The aim of the PCPT trial was to compare the incidence of prostate cancer in men treated with finasteride or placebo. The study protocol contains some methodological difficulties. Solutions were adopted designed to minimise damaging bias which could have incorrectly suggested that finasteride was beneficial, or masked a benefit of the drug. As finastéride reduced PSA by an average of 50% the PSA values were interpreted against a new threshold to provide as many biopsies in the finasteride arm as in the placebo arm. In view of the reduction in gland volume, the biopsy protocol was modified to prioritise lateral peripheral cores. Patient adherence to treatment in the finasteride arm and contamination of the placebo arm by people taking finasteride retrospectively after randomisation were taken into account. The increase in sensitivity of rectal examination and in the number of prostatic resections in the placebo arm were also two sources of bias in favour of finasteride. The different envisaged sources of bias were analysed and plans were adopted to manage these wherever possible. Analysis of the sources of bias led to the conclusion that systematic end of trial biopsies constituted the most robust end point.


Does viewing of surgical specimen improve patient satisfaction? A cohort study.
The purpose of this study was to assess a cohort of patients who underwent elective gynecologic surgery to determine whether viewing of intraoperative photographic images by patients played a role in patient satisfaction. Three hundred and eighteen patients participated in this study, out of which 159 patients were assigned to view the operative photographic pictures and 159 were not. According to the results obtained in this study, a greater number of women who viewed their operative photographic pictures, expressed satisfaction in comparison to those who did not. In addition, this difference was significantly greater in those who encountered intra- and/or postoperative complications.


Malnutrition and bilateral central retinal vein occlusion in a young woman: a case report.
Can vitamin B12 and folate deficiency cause central retinal vein occlusion? We conducted a literature search to find out whether nutritional deficiency of vitamin B12 and folate can lead to impaired vision. The patient in the article presented in an eye-casualty department in the North East of England with gradual painless visual loss over six weeks. She was found to have bilateral central retinal vein occlusion with significant anaemia and vitamin B12 and folate deficiency. Vitamin B12 and folate deficiency can lead to elevated levels of homocysteine. We found a large amount of published data relating central retinal vein occlusion to elevated homocysteine levels, but there was a lack of conclusive evidence for this association Patients should be asked about their dietary history where a thrombotic event is suspected or confirmed.


Endostatin inhibits the growth of endometriotic lesions but does not affect fertility.
To determine whether endometriosis can be treated with the angiogenesis inhibitor endostatin and the effect of this treatment on fertility and reproduction. Pharmacologic intervention in a surgically induced model of endometriosis and in female mice undergoing mating. Animal research facility. Eight-week-old, female C57BL/6 and SCID mice. After implantation of autologous endometrium, mice received endostatin or the vehicle-matched control for 4 weeks. For the reproductive function study, mice receiving endostatin or vehicle were mated and reproductive functions were observed. Growth of endometriotic lesions after 4 weeks of treatment; estrous cycling, corpus luteum formation, serum hormone levels, and mating time as fertility measures; and pregnancy rates, length of pregnancy, fetal vitality, number, and outcome of litter as reproductive measures. Endostatin suppressed the growth of endometriotic lesions by 47% compared with controls. Estrous cycling and corpus luteum formation were normal in both groups. Female mice receiving endostatin were as fertile as mice receiving vehicle, had normal pregnancies, and delivered the same number of pups. The offspring were healthy without teratogenic stigmata and reproduced normally themselves. Antiangiogenic therapy with endostatin may present a promising novel, nontoxic therapeutic option for patients with endometriosis.


Cardiovascular effects of dobutamine in severe congestive heart failure.
The effects of continuous infusion of dobutamine 5 to 15 microgram/Kg./min. were studied in 17 patients using right heart catheterizations, echocardiography, and/or the Systolic Time Intervals. HR increase was dose-related, but insignificant (p less than 0.05) rate increase was obtained at infusion rates below 15 microgram/Kg.)min. C.O. increased from 2.9 +/- 0.7 to 5.0 +/- 1.2 liters/min. (p less than 0.001), and the stroke volume from 30 +/- 6 to 49 +/- 14 ml./min. (p less than 0.005). The mean BP did not change, P.A.W.P. decreased from 30 +/- 7 to 20 +/- 8 mm. Hg (p less than 0.001) and R.A.P. from 20.0 to 12.0 mm. Hg (p less than 0.005). The P.E.P.I. decreased from 160.93 +/- 54.91 to 133.4 +/- 28.7 msec. (p less than 0.050). Echo-determined mean VCf increased from 0.387 +/- 0.14 to 0.537 +/- 0.13 cm. (p less than 0.010), diastolic diameter did not change significantly, but the end systolic diameter decreased from 6.020 +/- 0.69 to 5.750 +/- 0.70 cm. (p less than 0.025). During a mean infusion period of 75 hours, the only side effects noted were transient nausea and/or vomiting at 15 microgram/Kg./min. dose range in two patients, and multifocal P.V.C.'s following 68 hours of infusion in another patient. It is concluded that in the dose range of 5 to 15 microgram/Kg./min., dobutamine is well tolerated and is a very potent inotropic agent with only minor effects on the heart rate and blood pressure.


Collapsing ROC approach for risk prediction research on both common and rare variants.
Risk prediction that capitalizes on emerging genetic findings holds great promise for improving public health and clinical care. However, recent risk prediction research has shown that predictive tests formed on existing common genetic loci, including those from genome-wide association studies, have lacked sufficient accuracy for clinical use. Because most rare variants on the genome have not yet been studied for their role in risk prediction, future disease prediction discoveries should shift toward a more comprehensive risk prediction strategy that takes into account both common and rare variants. We are proposing a collapsing receiver operating characteristic (CROC) approach for risk prediction research on both common and rare variants. The new approach is an extension of a previously developed forward ROC (FROC) approach, with additional procedures for handling rare variants. The approach was evaluated through the use of 533 single-nucleotide polymorphisms (SNPs) in 37 candidate genes from the Genetic Analysis Workshop 17 mini-exome data set. We found that a prediction model built on all SNPs gained more accuracy (AUC = 0.605) than one built on common variants alone (AUC = 0.585). We further evaluated the performance of two approaches by gradually reducing the number of common variants in the analysis. We found that the CROC method attained more accuracy than the FROC method when the number of common variants in the data decreased. In an extreme scenario, when there are only rare variants in the data, the CROC reached an AUC value of 0.603, whereas the FROC had an AUC value of 0.524.


[Description of tuberculosis outbreak and usefulness of mediastinal ultrasound].
Describe a school outbreak of tuberculosis and compare the features with other outbreaks. Propose mediastinal ultrasound as an aid in its diagnosis and monitoring. Retrospective descriptive study of the transmission of tuberculosis infection in a micro-epidemic. Infection criteria: Mantoux ≥ 5mm, asymptomatic patient, with normal radiological, microbiological and analytical studies. Disease criteria: Mantoux ≥ 5mm and any of the above pathological studies. Mediastinal ultrasound was used as a complementary method of chest radiography (CXR). Computed tomography (CT) as the combined result of ultrasound and CXR was inconclusive. Seventeen out of 412 students were infected and 16 with tuberculosis disease. In addition, 4 out-school contacts were diagnosed of tuberculosis. one erythema nodosum, one Henoch-Schönlein purpura, twelve with cough and fever nine. CXR results in patients: 3 normal, 7 inconclusive, and 10 with the following findings: 2 pneumonia, 2 pulmonary atelectasis, 2 primary complexes, 4 patients only with lymphadenopathy. All patients with normal or indeterminate CXR showed mediastinal lymphadenopathy visualized on ultrasound but 3 demonstrated by CT. Microbiological isolation: 6 cases (31%). The number of infected patients was higher compared to other outbreaks There is great variability in the diagnostic approach to tuberculosis micro-epidemics. It is complex to differentiate between infection and disease. The clinical, laboratory and CXR are nonspecific. There are patients with normal CXR, pathological ultrasound and microbiological isolation. Mediastinal ultrasound can play an important role in the diagnosis and management of tuberculosis disease.


Shear bond strength between light polymerized hard reline resin and denture base resin subjected to long term water immersion.
The effect of long-term water immersion on the shear bond strength between denture base resin and Triad visible-light-polymerized (VLP) direct hard reline resin is not known. The aim of this study was to investigate the bonding characteristics of Triad VLP direct hard reline resin to heat-polymerized denture base resin subjected to long-term water immersion. Ninety circular disks, 15 mm in diameter and 3 mm thick, of denture base resin were polymerized from a gypsum mold. Sixty specimens were subjected to water immersion and 30 were stored at ambient room temperature for 4 months. Thirty water-immersed specimens were dried with gauze (group 1), while the other 30 water-immersed specimens were dried with a hair dryer (group 2). The dry specimens (n = 30) represented the control group (group 3). All specimens were air abraded and painted with bonding agent before packing Triad VLP direct hard-reline resin. Specimens in each group were subjected to thermal cycling for 50,000 cycles between 4 °C and 60 °C water baths with 1-min dwell time at each temperature. The bond strength at which the bond failed under stress was recorded using a universal testing machine. One-way ANOVA and Tukey post hoc comparison were applied to find significant differences between groups (α = 0.05). Significant differences in mean shear bond strength among the specimens existed because of variable water content in the denture base resin (P < 0.05). Group 3 (dry) was higher than group 2 (desiccated), and the lowest was group 1 (saturated). The shear bond strength of Triad VLP direct hard reline resin to denture base resin depended on the water content in the denture base resin. The dry denture base resin demonstrated superior bond strength compared with the desiccated and water-saturated denture base resins.


Characterization of bioactive compound isolated from Micromonospora marina KPMS1 and its activity against emerging antibiotics resistant strains of Klebsiella pneumoniae HAUTI7 and Proteus vulgaris HAUTI14.
Multiple antibiotic resistances have increased gradually in many classes of antibiotics among the gram negative organisms like Klebsiella pneumoniae and Proteus vulgaris which are the major causes of infection among worldwide. Nearly a hundred urine samples were collected, among them 16 urine samples were having plasmid and its resistant to various antibiotics. This present investigation has determined the resistant plasmid pattern of multi drug resistant Klebsiella pneumoniae and Proteus vulgaris from urinary tract site isolated from hospital patients. The detection and characterization of antimicrobial metabolite derived from marine sediments that produce potent activity against multidrug resistant pathogen. The 16S rRNA sequencing results and phylogeny showed that the resistant bacteria belong to the genera of Klebsiella pneumoniae HAUTI7 and Proteus vulgaris HAUTI14. The antibacterial activity and the characterization of bioactive compound like FT-IR and NMR studies were performed to analyze the structural elucidation of active compounds derived from marine source Micromonospora marina KPMS1. The 16S rRNA sequences of Micromonospora marina KPMS1was deposited in the Gen bank with the accession number MH036351. The effective bioactive compound derived from marine sediments are virtually unlimited interest that control the emerging multiple antibiotic resistant strains.


Implementation of violence prevention education in clinical settings.
Clinical settings provide excellent opportunities for educating adolescent patients about violence prevention. This paper reports on the experience of the Violence Prevention Project of Boston in implementing comprehensive violence prevention activities in different clinical settings. The project's evaluation revealed that staff training, specialized educational materials and staff commitment are required for both initial and sustained implementation of the education. Waiting room brochures and videos are inadequate to address the problem. Conversely, clinical settings can host a variety of educational experiences that generate significant response from patients, families and friends. Clinical settings can also be integrated into broader community-based efforts.


Molecular cloning and nucleotide sequence of human pancreatic secretory trypsin inhibitor (PSTI) cDNA.
We have isolated and sequenced a cDNA clone coding for the human pancreatic secretory trypsin inhibitor (PSTI) from a human pancreatic cDNA library. The predicted product consists of 79 amino acids, and contains no apparent functional polypeptide other than PSTI. Southern blot analysis suggests that there is one copy of PSTI gene per haploid genome. This gene seems to be expressed not only in pancreas, but also in gastric mucosa, since a Northern blot analysis demonstrated the presence of a poly(A) RNA of the same size as in the pancreas. A comparison of sequences between human PSTI mRNA and mouse epidermal growth factor (EGF) mRNA revealed a high homology, suggesting that they share a common ancestral DNA sequence.


Tissue-specific genome editing of laminA/C in the posterior silk glands of Bombyx mori.
The RNA-guided CRISPR/Cas9 system has been shown to be a powerful tool for genome editing in various organisms. A comprehensive toolbox for multiplex genome editing has been developed for the silkworm, Bombyx mori, a lepidopteran model insect of economic importance. However, as previous methods mainly relied on delivery of transient Cas9/guide RNA (gRNA), they could not be used in loss-of-function studies of essential genes. Here, we report a simple and versatile tissue-specific genome editing strategy. We perform a proof-of-principle demonstration by establishing and crossing two transgenic B. mori lines, one expressing Cas9 protein in the posterior silk glands (PSGs) and the other constitutively expressing BmlaminA/C (BmLMN) gRNA. All BmLMN alleles in the PSG cells were edited precisely at the target genome region, resulting in diverse mutations. mRNA expression of BmLMN was reduced by up to 75%, and only very low levels of BmLaminA/C protein were detected. Knockout of BmLMN produced obvious defects in gland cell development and cocoon production. In this study, we developed an efficient strategy for spatially controlled genome editing, providing unprecedented opportunities for investigating the function of essential/lethal genes in B. mori, with potential application for other insects.


Longer-term outcomes of radiofrequency ablation for locally recurrent papillary thyroid cancer.
The purpose of this study was to evaluate the longer-term efficacy of ultrasound (US)-guided radiofrequency ablation (RFA) for treating locally recurrent papillary thyroid cancer (PTC). We retrospectively reviewed 29 patients who underwent RFA for 46 recurrent PTC between September 2008 and April 2012 and were subsequently followed up for at least 5 years. Follow-up included size change on US and thyroglobulin (Tg) level at 1, 3, 6, and 12 months and every 6-12 months thereafter. Any complications were reported during follow-up. The mean follow-up duration after RFA was 80 ± 17.3 months (range, 60-114 months). Tumor volume decreased significantly, from 0.25 ± 0.42 mL before ablation to 0.01 ± 0.08 mL at the final evaluation (p < 0.001), with a mean volume reduction of 99.5% ± 2.9%. Forty-two of the 46 treated tumors (91.3%) had completely disappeared by the final evaluation. The mean serum Tg level decreased from 2.55 ± 4.7 to 0.75 ± 1.83 ng/dL (p < 0.001). There were no delayed complications associated with RFA during the follow-up period. RFA seems to be an effective minimally invasive therapy for the treatment of locally recurrent PTC even in the longer-term period. • RFA is an effective local control treatment option for recurrent PTCs even in the longer-term period with mean tumor VRR of 99.5% and the complete disappearance of the treated tumors in 91.3%. • The mean serum Tg level decreased significantly after RFA and biochemical remission rate was 51.7%. • No delayed complication after RFA for local recurrent PTC.


An in vivo investigation of the effect of anthraquinones on the turnover of aggrecans in spontaneous osteoarthritis in the guinea pig.
We present details of a method which allows for the determination of chondroitin sulphate turnover in vivo using the guinea pig. Such methods have been utilised to examine the effects of diacetyl rhein, a compound with purported anti-osteoarthritic activity, and several related anthraquinone analogues on the turnover of chondroitin. Since the guinea pig develops spontaneous osteoarthritis, this may give useful information on the potential for such compounds to inhibit the progression of osteoarthritis. The results show that several of the anthraquinones are capable of reducing the turnover of chondroitin 4- but not 6-sulphate. This may indicate potential mechanisms for the breakdown of guinea pig cartilage aggrecans. We propose that these techniques could be useful for the screening of chemical agents with useful activity against osteoarthritis.


Retrospective evaluation of inpatient celecoxib use after total hip and knee arthroplasty at a Veterans Affairs Medical Center.
A retrospective cohort study (1.5 years) was performed to investigate the efficacy of celecoxib vs non-celecoxib use in patient who underwent total knee arthroplasty (TKA) and total hip arthroplasty (THA). Study time frame encompassed a pre and post period of a local policy decision opening access to short-term celecoxib use after TKA/THA. Primary end point was the amount of opioid use during their inpatient stay postprocedure. The TKA (n = 81) and THA (n = 60) groups were analyzed independently. Both celecoxib groups used significantly less opioids during their inpatient stay vs noncelecoxib groups, given in oral morphine milligram equivalents (TKA: 203 vs 337 mg, P = .002; THA: 214 vs 336 mg, P = .005). Other secondary outcome measures showed that the celecoxib groups also reported reduction in pain scores, total as needed (PRN) opioid doses, PRN opioid doses per day, average dose of PRN opioids, total PRN opioids, use of intravenous opioids, and rehabilitation facility admissions (in the TKA group only). Linear regression analysis showed a statistically significant inverse relationship between opioid consumption and age. Short-term celecoxib use after TKA/THA may lead to a reduction in overall opioid use and improved pain scores; however, further studies will be required to validate the results of this study.


Expression of orexin-A and orexin receptors in the kidney and the presence of orexin-A-like immunoreactivity in human urine.
Orexin-A (hypocretin-1), a neuropeptide with stimulatory actions on arousal and appetite, was originally shown to be specifically expressed in the hypothalamus. We studied expression of orexin-A and orexin receptors in the kidney and the presence of orexin-A-like immunoreactivity in human urine. Immunocytochemistry showed that orexin-A-like immunoreactivity and two types of orexin receptors (types 1 and 2) were localized in the tubules of the human kidney obtained at autopsy. Orexin-A-like immunoreactivity was detected in human kidneys (21.3 +/- 6.2 fmol/g wet weight, mean +/- S.E.M., n = 4) and rat kidneys (16.2 +/- 1.6 fmol/g wet weight, n = 5) by radioimmunoassay, although the levels were much lower than the levels in the brain. Orexin-A-like immunoreactivity was present in the urine obtained from male healthy volunteers (67.8 +/- 4.5 pmol/l, n = 5). Reverse phase high-performance liquid chromatography showed that most of orexin-A-like immunoreactivity of the urine extract was eluted earlier than authentic orexin-A, suggesting that orexin-A-like immunoreactivity in urine was modified to hydrophilic forms. Reverse transcriptase polymerase chain reaction showed expression of orexin receptors 1 and 2 mRNAs in the human kidney. These findings suggest that orexin-A is produced by the renal tubular cells and secreted into urine. Orexin-A may act on the kidney in the autocrine or paracrine fashion, or via the urine (urocrine fashion).