Building on the Ccr4-Not architecture.
In a recent issue of Nature Communications Ukleja and co-workers reported a cryo-EM 3D reconstruction of the Ccr4-Not complex from Schizosaccharomyces pombe with an immunolocalization of the different subunits. The newly gained architectural knowledge provides cues to apprehend the functional diversity of this major eukaryotic regulator. Indeed, in the cytoplasm alone, Ccr4-Not regulates translational repression, decapping and deadenylation, and the Not module additionally plays a positive role in translation. The spatial distribution of the subunits within the structure is compatible with a model proposing that the Ccr4-Not complex interacts with the 5' and 3' ends of target mRNAs, allowing different functional modules of the complex to act at different stages of the translation process, possibly within a circular constellation of the mRNA. This work opens new avenues, and reveals important gaps in our understanding regarding structure and mode of function of the Ccr4-Not complex that need to be addressed in the future.


Fosamprenavir: advancing HIV protease inhibitor treatment options.
Fosamprenavir, the prodrug formulation of amprenavir, is a protease inhibitor recently approved in the US for the treatment of HIV infection. This agent combines the pharmacological profile of amprenavir with a low pill burden and flexible dosing schedule. Three large international trials have been completed. In treatment-naive patients, fosamprenavir, both ritonavir-boosted and -unboosted, met primary end points of non-inferiority against nelfinavir. Among naive patients, no protease inhibitor mutations emerged in those failing the boosted fosamprenavir regimen. Boosted fosamprenavir was compared to lopinavir/ritonavir in treatment-experienced patients. Non-inferiority was not achieved but similar numbers of patients achieved viral suppression when fosamprenavir was dosed twice-daily. Fosamprenavir demonstrates a favourable lipid profile in naive patients, and a low incidence of adverse effects. Fosamprenavir, as with lopinavir/ritonavir, distinguishes itself among other protease inhibitors with its potent activity in those with advanced HIV. Future trials comparing fosamprenavir with current standard regimens will further assist in defining its role in clinical practice.


Molecular analysis of plasmid DNA repair within ultraviolet-irradiated Escherichia coli. I. T4 endonuclease V-initiated excision repair.
The process by which DNA-interactive proteins locate specific sequences or target sites on cellular DNA within Escherichia coli is a poorly understood phenomenon. In this study, we present the first direct in vivo analysis of the interaction of a DNA repair enzyme, T4 endonuclease V, and its substrate, pyrimidine dimer-containing plasmid DNA, within UV-irradiated E. coli. A pyrimidine dimer represents a small target site within large domains of DNA. There are two possible paradigms by which endonuclease V could locate these small target sites: a processive mechanism in which the enzyme "scans" DNA for dimer sites or a distributive process in which dimers are located by random three-dimensional diffusion. In order to discriminate between these two possibilities in E. coli, an in vivo DNA repair assay was developed to study the kinetics of plasmid DNA repair and the dimer frequency (i.e. the number of dimer sites on a given plasmid molecule) in plasmid DNA as a function of time during repair. Our results demonstrate that the overall process of plasmid DNA repair initiated by T4 endonuclease V (expressed from a recombinant plasmid within repair-deficient E. coli) occurs by a processive mechanism. Furthermore, by reducing the temperature of the repair incubation, the endonuclease V-catalyzed incision step has been effectively decoupled from the subsequent steps including repair patch synthesis, ligation, and supercoiling. By this manipulation, it was determined that the overall processive mechanism is composed of two phases: a rapid processive endonuclease V-catalyzed incision reaction, followed by a slower processive mechanism, the ultimate product of which is the dimer-free supercoiled plasmid molecule.


Phosphorus-Containing Food Additives in the Food Supply-An Audit of Products on Supermarket Shelves.
Phosphorus (P)-containing food additives pose a risk for chronic kidney disease (CKD) patients. We aimed to investigate the prevalence of P-containing additives in the Finnish food supply across different food categories to evaluate their burden in CKD, reflecting the situation in Europe. The dataset of 6,176 products was obtained in June-August 2019 from the foodie.fi website, which contains all foodstuffs sold in the grocery stores of the S Group (46% of the Finnish market share in 2019). The food category, full product name, type of P additive (inorganic, organic, and natural P-containing), and reporting methods (name or E number) of P additives were recorded. Duplicates and products lacking ingredient information were excluded. The prevalence of P additives was 36% in the final sample (n = 5,149). Among food categories, the prevalence varied from 4% in dairy-based snacks to 67% in meat products. Altogether 17 different P additives were observed. Inorganic P additives were the most common P additive type, present in 20% of foodstuffs. Natural P-containing additives were observed in 19% and organic P additives in 2% of foodstuffs. The most commonly used P additives were lecithin (E 322), pyrophosphate (E 450), and triphosphate (E 451). E number was used as a reporting method in 49% of foodstuffs, and full name in 44% of foodstuffs. Reporting by E number was particularly common in the products containing inorganic P. The use of P additives is common in the Finnish food supply, indicating the situation in Europe. The high prevalence of inorganic, that is, the most absorbable and potentially most harmful P additives in particular food groups, and their usual reporting only by E numbers can create challenges in CKD dietary counseling.


International monitoring of capacity of treatment systems for alcohol and drug use disorders: Methodology of the Service Capacity Index for Substance Use Disorders.
We aimed to develop a Service Capacity Index for Substance Use Disorders (SCI-SUD) that would reflect the capacity of national health systems to provide treatment for alcohol and drug use disorders, in terms of the proportion of available service elements in a given country from a theoretical maximum. Data were collected through the WHO Global Survey on Progress with Sustainable Development Goals (SDG) Health Target 3.5, conducted between December 2019 and July 2020 to produce the SCI-SUD, based on 378 variables overall. The SCI-SUD was directly derived for 145 countries. We used multiple imputation to produce comparable SCI-SUD estimates for countries that did not submit data (40 countries) or had very high level of missingness (9 countries). The final SCI-SUD demonstrates considerable consistency and internal stability and is strongly associated with the macro-level economic, healthcare-related and epidemiologic (such as prevalence rates) variables. The presented methodology represents a step forward in monitoring the global situation in regard to the development of treatment systems for SU disorders, however, further work is warranted to improve the external validity of the measure (e.g., in-depth data generation in countries) and ensure its feasibility for regular reporting (e.g., reducing the number of variables).


Colesevelam hydrochloride in clinical practice: a new approach in the treatment of hypercholesterolaemia.
Hypercholesterolaemia is a major risk factor for atherosclerosis and coronary heart disease. Treatment with lipid lowering agents reduces the risk of vascular events. Colesevelam is a novel bile acid sequestrant (BAS) indicated for the treatment of hypercholesterolaemia, either as monotherapy or in combination with statins. This article reviews the efficacy, tolerability and safety of colesevelam in clinical practice. The literature search was based on a PubMed search up to January 2008. Colesevelam, used alone or in combination with other hypolipidaemic agents (statins, ezetimibe and fenofibrate), has an overall favourable effect on lipid profile. Specifically, colesevelam reduces total and low-density lipoprotein cholesterol (LDL-C) and apolipoprotein B levels and increases high-density lipoprotein cholesterol and apolipoprotein AI. However, colesevelam may slightly raise triglyceride levels. Colesevelam can improve glycaemic control in diabetic patients. Moreover, it may have anti-inflammatory properties, as it can reduce high sensitivity C-reactive protein concentration. Colesevelam almost lacks the intense side effects of previously used BASs, thus resulting in better patient compliance. However, the dose regimen consisting of up to 7 tablets/day and high cost may limit its use. Colesevelam is a safe alternative for those intolerant to other lipid lowering medication. This BAS also provides an option for patients who do not reach their LDL-C goal despite treatment with a statin.


Repurposing of existing FDA approved drugs for Neprilysin inhibition: An in-silico study.
Neprilysin (NEP) is a neutral endopeptidase with diverse physiological roles in the body. NEP's role in degradation of diverse classes of peptides such as amyloid beta, natriuretic peptide, substance P, angiotensin, endothelins, etc., is associated with pathologies of alzheimer's, kidney and heart diseases, obesity, diabetes and certain malignancies. Hence, the functional inhibition of NEP in the above systems can be a good therapeutic target. In the present study, in-silico drug repurposing approach was used to identify NEP inhibitors. Molecular docking was carried out using GLIDE tool. 2934 drugs from the ZINC12 database were screened using high throughput virtual screening (HTVS) followed by standard precision (SP) and extra precision (XP) docking. Based on the XP docking score and ligand interaction, the top 8 hits were subjected to free ligand binding energy calculation, to filter out 4 hits (ZINC000000001427, ZINC000001533877, ZINC000000601283, and ZINC000003831594). Further, induced fit docking-standard precision (IFD-SP) and molecular dynamics (MD) studies were performed. The results obtained from MD studies suggest that ZINC000000601283-NEP and ZINC000003831594-NEP complexes were most stable for 20ns simulation period as compared to ZINC000001533877-NEP and ZINC000000001427-NEP complexes. Interestingly, ZINC000000601283 and ZINC000003831594 showed similarity in binding with the reported NEP inhibitor sacubitrilat. Findings from this study suggest that ZINC000000601283 and ZINC000003831594 may act as NEP inhibitors. In future studies, the role of ZINC000000601283 and ZINC000003831594 in NEP inhibition should be tested in biological systems to evaluate therapeutic effect in NEP associated pathological conditions.


Can eggs in a cavity be a female secondary sexual signal? Male nest visits and modelling of egg visual discrimination in blue tits.
Eggshell colouration is thought to function as a female-specific secondary sexual trait. While tests of this idea are rapidly accumulating in cavity-nesting birds, some fundamental underlying assumptions remain rarely investigated: namely, can males see eggshell coloration and perceive colour differences between the eggs of different females? We tested these two key assumptions in a natural population of blue tits (Cyanistes caeruleus). Using transponders, we tracked male nest visits and found that all males visited their nest-boxes while eggs were present and often visually accessible. Interestingly, some males also visited neighbouring nests. We then tested whether birds could detect eggshell coloration using models of avian colour vision; models were performed with and without limitations on visual performance owing to dim light. Both models found that differences in eggshell brightness were often easier to discriminate than differences in colour; there was more contrast in white eggshell background between clutches than within and its contrast against nest background was repeatable within clutches, suggesting these features could act as signals. Yet, the detectability of these contrasts depended entirely on model assumptions of visual limitations. Consequently, we need a better understanding of underlying visual mechanisms in dim-light environments and behavioural discrimination experiments before confirming the signalling potential of eggshell coloration.


Mutation scanning-based analysis of Theileria orientalis populations in cattle following an outbreak.
Bovine theileriosis is a tick-borne disease caused by one or more hemoprotozoan parasites of the genus Theileria. In the past, Theileria infection in cattle in Australia was largely asymptomatic and recognized to be associated with Theileria buffeli. However, outbreaks of theileriosis have occurred in beef and dairy cattle in subtropical climatic regions (New South Wales) of Australia. There is also one published report of a recent theileriosis outbreak in a beef farm near Seymour in the southeastern state of Victoria. In order to gain an improved insight into the genetic composition of Theileria populations following this outbreak, we undertook herein an integrated PCR-coupled mutation scanning-sequencing-phylogenetic analysis of sequence variation in part of the major piroplasm surface protein (MPSP) gene within and among samples from cattle involved in the outbreak. Theileria DNA was detected in 89.4% of 94 cattle in the Seymour farm; the genetic analysis showed that the ikeda and chitose genotypes representing the Theileria orientalis complex were detected in 75 and 4.8% of 84 infected cattle, respectively, and that mixed populations of these two genotypes were found in 20.2% of infected cattle. Given unpublished reports of a significant increase in the number of outbreaks in Victoria, future investigations should focus sharply on elucidating the epidemiology of Theileria to subvert the economic impact on the cattle industry in this state. Although used here to explore genetic variation within the T. orientalis complex in Australia, a mutation scanning-based approach has broad applicability to other species of Theileria in other countries.


Image-based method for retrospective correction of physiological motion effects in fMRI: RETROICOR.
Respiration effects and cardiac pulsatility can induce signal modulations in functional MR image time series that increase noise and degrade the statistical significance of activation signals. A simple image-based correction method is described that does not have the limitations of k-space methods that preclude high spatial frequency correction. Low-order Fourier series are fit to the image data based on time of each image acquisition relative to the phase of the cardiac and respiratory cycles, monitored using a photoplethysmograph and pneumatic belt, respectively. The RETROICOR method is demonstrated using resting-state experiments on three subjects and compared with the k-space method. The method is found to perform well for both respiration- and cardiac-induced noise without imposing spatial filtering on the correction. Magn Reson Med 44:162-167, 2000.


Transanal resection in the palliation of rectal cancer.
Transanal resection of rectal tumours employs a technique similar to transurethral resection of the prostate, using a modified urological resectoscope with continuous fluid irrigation. It is applicable to locally advanced or recurrent rectal cancers, villous adenomas, and some early carcinomas. The technique avoids interference with anal sphincter function and is well tolerated by unfit and elderly patients. It provides good relief from tenesmus, bleeding, diarrhoea, mucous discharge, and obstruction, and is readily repeatable. Transanal resection is a useful addition to conventional techniques in the treatment of rectal cancer.


Guidelines for the treatment of severe sepsis and septic shock - management of the infectious agent - diagnosis.
Sepsis is a common and lethal condition that carries a substantial financial burden and is the primary cause of death in intensive care units. Early diagnosis and treatment of patients has been clearly shown to improve prognosis. Therefore, early diagnosis of infections and control of the primary infection site are fundamental to improving patients' prognosis. This guideline reviews the available evidence concerning the primary strategies for the diagnosis of infection.


Maintaining tissue perfusion in high-risk surgical patients: a systematic review of randomized clinical trials.
Surgical patients with limited organic reserve are considered high-risk patients and have an increased perioperative mortality. For this reason, they need a more rigorous perioperative protocol of hemodynamic control to prevent tissue hypoperfusion. In this study, we systematically reviewed the randomized controlled clinical trials that used a hemodynamic protocol to maintain adequate tissue perfusion in the high-risk surgical patient. We searched MEDLINE, Embase, LILACS, and Cochrane databases to identify randomized controlled clinical studies of surgical patients studied using a perioperative hemodynamic protocol of tissue perfusion aiming to reduce mortality and morbidity; the latter characterized at least one dysfunctional organ in the postoperative period. Pooled odds ratio (POR) and 95% confidence interval (CI) were calculated for categorical outcomes. Thirty-two clinical trials were selected, comprising 5056 high-risk surgical patients. Global meta-analysis showed a significant reduction in mortality rate (POR: 0.67; 95% CI: 0.55-0.82; P < 0.001) and in postoperative organ dysfunction incidence (POR: 0.62; 95% CI: 0.55-0.70; P < 0.00,001) when a hemodynamic protocol was used to maintain tissue perfusion. When the mortality rate was >20% in the control group, the use of a hemodynamic protocol to maintain tissue optimization resulted in a further reduction in mortality (POR: 0.32; 95% CI: 0.21-0.47; P < 0.00,001). Monitoring cardiac output with a pulmonary artery catheter and increasing oxygen transport and/or decreasing consumption also significantly reduced mortality (POR: 0.67; 95% CI: 0.54-0.84; P < 0.001 and POR: 0.71; 95% CI: 0.57-0.88; P < 0.05, respectively). Therapy directed at increasing mixed or central venous oxygen saturation did not significantly reduce mortality (POR: 0.68; 95% CI: 0.22-2.10; P > 0.05). The only study using lactate as a marker of tissue perfusion failed to demonstrate a statistically significant reduction in mortality (OR: 0.33; 95% CI: 0.07-1.65; P > 0.05). In high-risk surgical patients, the use of a hemodynamic protocol to maintain tissue perfusion decreased mortality and postoperative organ failure. Monitoring cardiac output calculating oxygen transport and consumption helped to guide therapy. Additional randomized controlled clinical studies are necessary to analyze the value of monitoring mixed or central venous oxygen saturation and lactate in high-risk surgical patients.


Isoniazid resistance profile and associated levofloxacin and pyrazinamide resistance in rifampicin resistant and sensitive isolates/from pulmonary and extrapulmonary tuberculosis patients in Pakistan: A laboratory based surveillance study 2015-19.
Pakistan is among top five high burden countries for tuberculosis and drug resistant TB. Among rifampicin sensitive new pulmonary TB (PTB), prevalence of isoniazid resistance is 8.3% (95%CI: 7.0-10.7) and resistance to fluoroquinolone is higher (11·1%, 95%CI: 7·8-14·3) than isoniazid resistance. Five year retrospective data (2015-2019) of drug susceptibility testing (DST) for Mycobacterium tuberculosis isolates, performed using recommended phenotypic (pDST) and/or genotypic (gDST) methods was analyzed stratified by rifampicin results for isoniazid resistance profiles and associated levofloxacin and pyrazinamide resistance. DST data was analyzed from 11045 TB patients. Isolates were tested using pDST (87%), gDST (92%) and both methods (79.5%). For both rifampicin and isoniazid, a significant difference (P < .001) was noted between resistance detected by pDST and gDST. Among isolates, tested by both methods (8787), 49% were resistant to rifampicin and 51.7% to isoniazid with discordance in resistant results of 15.8% for each, with 13.2% (570) of rifampicin resistance reported sensitive by pDST and 14.2% (660) of isoniazid resistance missed by gDST. Estimated isoniazid resistance among rifampicin sensitive new PTB, extrapulmonary TB and previously treated PTB was 9.8% (95%CI: 8.7-11.1), 6.8% (95%CI: 5.4-8.5) and 14.6% (95%CI: 11.8-17.9) respectively. Significant differences were reported between the genotypic profile of isoniazid resistance associated with rifampicin-resistant and sensitive isolates including detectable mutations (87% vs 71.6%), frequency of inhA (7.6% and 30.2%) and katG mutations (76.1% vs 41.2%) respectively. Among rifampicin resistant and sensitive isolates, a significantly higher level of resistance to levofloxacin and pyrazinamide was seen associated with isoniazid resistance. There are risks and many challenges in implementing WHO recommended treatment for isoniazid resistant tuberculosis. The laboratory based surveillance can complement random surveys in country specific planning for TB diagnostics and appropriate treatment regimens.


INTERMAP: the dietary data--process and quality control.
The aim of this report is to describe INTERMAP standardized procedures for assessing dietary intake of 4680 individuals from 17 population samples in China, Japan, UK and USA: Based on a common Protocol and Manuals of Operations, standardized collection by centrally trained certified staff of four 24 h dietary recalls, two timed 24-h urines, two 7-day histories of daily alcohol intake per participant; tape recording of all dietary interviews, and use of multiple methods for ongoing quality control of dietary data collection and processing (local, national, and international); one central laboratory for urine analyses; review, update, expansion of available databases for four countries to produce comparable data on 76 nutrients for all reported foods; use of these databases at international coordinating centres to compute nutrient composition. Chinese participants reported 2257 foods; Japanese, 2931; and UK, 3963. In US, use was made of 17,000 food items in the online automated Nutrition Data System. Average time/recall ranged from 22 min for China to 31 min for UK. Among indicators of dietary data quality, coding error rates (from recoding 10% random samples of recalls) were 2.3% for China, 1.4% for Japan, and UK; an analogous US procedure (re-entry of recalls into computer from tape recordings) also yielded low discrepancy rates. Average scores on assessment of taped dietary interviews were high, 40.4 (Japan) to 45.3 (China) (highest possible score: 48); correlations between urinary and dietary nutrient values--similar for men and women--were, for all 4680 participants, 0.51 for total protein, range across countries 0.40-0.52; 0.55 for potassium, range 0.30-0.58; 0.42 for sodium, range 0.33-0.46. The updated dietary databases are valuable international resources. Dietary quality control procedures yielded data generally indicative of high quality performance in the four countries. These procedures were time consuming. Ongoing recoding of random samples of recalls is deemed essential. Use of tape recorded dietary interviews contributed to quality control, despite feasibility problems, deemed remediable by protocol modification. For quality assessment, use of correlation data on dietary and urinary nutrient values yielded meaningful findings, including evidence of special difficulties in assessing sodium intake by dietary methods.


Type II collagen induced arthritis in rats: clinical and immunological studies.
Immunization of laboratory rats with native Type II collagen induces arthritis in approximately 40 percent of the individuals. Histological and radiological analysis have suggested that the lesion bears several similarities to human rheumatoid arthritis. Clinical studies conducted with this model indicate that it is responsive to the treatment with clinically-used NSAID'S, steroids and immunosuppressive agents. When rats with collagen arthritis were treated with DMARD's, only those treated with D-penicillamine showed clinical improvement (radiological evaluation). Type II collagen induced arthritis is complement-dependent and is an example of an immune complex mediated injury. Thus, passive arthritis can be induced in rats by the intravenous administration of affinity-purified anticollagen IgG. The passive lesion is transient and the administered IgG is detected on the articular cartilage of the hind paws. This articular cartilage also contains complement C3. Passive arthritis is also complement-dependent. Suppression of Type II collagen arthritis can be induced by the prior intravenous treatment of rats with either Type II collagen or its constituent peptide alpha 1, (II) CB10. Antigen-induced arthritis cannot be induced in those rats that have recovered from passive arthritis.


CpG-DNA protects against a lethal orthopoxvirus infection in a murine model.
CpG-DNA has been described as a potent activator of the innate immune system, with potential to protect against infection caused by a range of pathogens in a non-specific manner. Here two classes of CpG-DNA (CpG-A and CpG-B) have been investigated for their abilities to protect mice from infection with an orthopoxvirus (vaccinia virus). Dosing with either CpG-A or B by the intraperitonal or intranasal route protected mice against a subsequent intranasal challenge with vaccinia virus. To our knowledge, this is the first time CpG-mediated protection has been demonstrated at the lung surface. The level of protection was greater when CpG-DNA was administered intranasally demonstrating a clear relationship between the route of CpG dosing and infection route. Treatment with CpG-B reduced viral titer in the lung by 10,000-fold at day 3 post-infection. The CC chemokines RANTES and MIP-1beta were elevated in the broncho-alveolar lavage from animals treated intranasally with CpG-B compared to untreated and intraperitoneally dosed controls, and it is possible that these chemokines play a role in the clearance of intranasally delivered vaccinia virus.


Effect of dipyridamole and aspirin on late vein-graft patency after coronary bypass operations.
To study the prevention of occlusion of aortocoronary-artery bypass grafts, we concluded a prospective, randomized, double-blind trial comparing long-term administration of dipyridamole (begun two days before operation) plus aspirin (begun seven hours after operation) with placebo in 407 patients. Results at one month showed a reduction in the rate of graft occlusion in patients receiving dipyridamole and aspirin. At vein-graft angiography performed in 343 patients (84 per cent) 11 to 18 months (median, 12 months) after operation, 11 per cent of 478 vein-graft distal anastomoses were occluded in the treated group, and 25 per cent of 486 were occluded in the placebo group. The proportion of patients with one or more distal anastomoses occluded was 22 per cent of 171 patients in the treated group and 47 per cent of 172 in the placebo group. All grafts were patent within a month of operation in 94 patients in the placebo group and 116 patients in the treated group; late development of occlusions was reduced from 27 per cent in the placebo group to 16 per cent in the treatment group. The results show that dipyridamole and aspirin continue to be effective in preventing vein-graft occlusion late after operation, and we believe that such treatment should be continued for at least one year.


Is Global Warming likely to cause an increased incidence of Malaria?
The rise in the average temperature of earth has been described as global warming which is mainly attributed to the increasing phenomenon of the greenhouse effect. It is believed that global warming can have several harmful effects on human health, both directly and indirectly. Since malaria is greatly influenced by climatic conditions because of its direct relationship with the mosquito population, it is widely assumed that its incidence is likely to increase in a future warmer world.This review article discusses the two contradictory views regarding the association of global warming with an increased incidence of malaria. On one hand, there are many who believe that there is a strong association between the recent increase in malaria incidence and global warming. They predict that as global warming continues, malaria is set to spread in locations where previously it was limited, due to cooler climate. On the other hand, several theories have been put forward which are quite contrary to this prediction. There are multiple other factors which are accountable for the recent upsurge of malaria: for example drug resistance, mosquito control programs, public health facilities, and living standards.


Recent methods for assessing osteoporosis and fracture risk.
In the management and treatment of osteoporosis, the target is to assess fracture risk and the end-point is to prevent fractures. Traditionally, measurement of bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA) has been the standard method for diagnosing osteoporosis, in addition to assessing fracture risk and therapeutic effects. Quantitative computed tomography (QCT) can quantify volumetric BMD, and cancellous bone can be measured independently of surrounding cortical bone and aortic calcification. Hip structure analysis (HSA) is a method using the DXA scan image and provides useful data for assessing hip fracture risk. Recently, new tools to assess osteoporosis and fracture risk have been developed. One of the recent advances has been the development of the FRAX (Fracture Risk Assessment Tool), which is helpful in conveying fracture risk to patients and providing treatment guidance to clinicians. Another advance is the finite element (FE) method based on data from computed tomography (CT), which is useful for assessing bone strength, fracture risk, and therapeutic effects on osteoporosis. In selecting the most appropriate drug for osteoporosis treatment, assessment by bone metabolic markers is an important factor. In this review, recent patents for assessing osteoporosis and fracture risk are discussed.


Anthropometric and physical performance characteristics of top-elite, elite and non-elite youth female team handball players.
In order to maximise the potential for success, developing nations need to produce superior systems to identify and develop talent, which requires comprehensive and up-to-date values on elite players. This study examined the anthropometric and physical characteristics of youth female team handball players (16.07 ± 1.30 years) in non-elite (n = 47), elite (n = 37) and top-elite players (n = 29). Anthropometric profiling included sum of eight skinfolds, body mass, stature, girths, breadths and somatotype. Performance tests included 20 m sprint, counter-movement jump, throwing velocity, repeated shuttle sprint and jump ability test, and Yo-Yo Intermittent Recovery Test Level 1. Youth top-elite players had greater body mass, lean mass, stature, limb girths and breadths than elite and non-elite players, while only stature and flexed arm were higher in elite compared to non-elite players (all P < 0.05). Sum of skinfolds and waist-to-hip ratio were similar between groups (P > 0.05). Top-elite performed better in most performance tests compared to both elite and non-elite players (P < 0.05), although maximal and repeated 10 m sprints were similar between playing standards (P > 0.05). Elite outperformed non-elite players only in throwing velocity. The findings reveal that non-elite players compare unfavourably to top-elite international European players in many anthropometric and performance characteristics, and differ in a few characteristics compared to elite European club team players. This study is useful for emerging team handball nations in improving talent identification processes.


[Surgical treatment for left-sided heart valve disease in a national reference hospital in Lima, Peru].
To describe the preoperative, intraoperative and postoperative characteristics of patients with left-sided heart valve disease treated in the thoracic and cardiovascular surgery service of a national reference hospital; as well as to describe the occurrence of thromboembolic and bleeding events in these patients. A retrospective longitudinal study was carried out, which included 185 patients who underwert surgery between 1999 and 2006 at the Hospital Nacional Dos de Mayo (Lima, Peru). The patients were divided into 4 groups: patients with mitral commissurotomy; with aortic valve replacement; with mitral valve replacement and with double valve replacement. T-student test, Chi-square test, analysis of variance and Bonferroni test were used. The survival analysis took into account the severe thromboembolic and bleeding events and a follow-up period of 7 years was set. The average durations of disease was 4.6 years. The most frequent etiology was rheumatic valve disease (74.6%). Hospital mortality was 3.8%, the most frequent cause of death was low cardiac output syndrome associated to multiple organ failure. The incidence of ischemic events (thrombosis or embolization) in patients who had a long-term valve replacement (more than 6 months) was 3.2%, and the incidence of bleeding events was 4.3%. The results of surgical treatment for left-sided heart valve disease in Peru are favorable. The rate of complications and hospital mortality rate, as well as the long-term thromboembolic and bleeding events are comparable to those reported in the world literature.


Ubiquity of Superconducting Domes in the Bardeen-Cooper-Schrieffer Theory with Finite-Range Potentials.
Based on recent progress in mathematical physics, we present a reliable method to analytically solve the linearized Bardeen-Cooper-Schrieffer (BCS) gap equation for a large class of finite-range interaction potentials leading to s-wave superconductivity. With this analysis, we demonstrate that the monotonic growth of the superconducting critical temperature T_{c} with the carrier density n predicted by standard BCS theory, is an artifact of the simplifying assumption that the interaction is quasilocal. In contrast, we show that any well-defined nonlocal potential leads to a "superconducting dome," i.e., a nonmonotonic T_{c}(n) exhibiting a maximum value at finite doping and going to zero for large n. This proves that, contrary to conventional wisdom, the presence of a superconducting dome is not necessarily an indication of competing orders, nor of exotic superconductivity.


Reliability of shoulder range of motion comparing a goniometer to a digital level.
The clinical use of digital levels, for joint measurement, may be a viable alternative to standard goniometry. The purpose of this study was to determine the intra- and intertester reliability of a construction grade digital level compared to the standard universal goniometer for measurements for active assisted shoulder range of motion (ROM). Two experienced physical therapists measured shoulder flexion, external rotation (ER), and internal rotation (IR) ROM bilaterally, on two different occasions, in 20 patients (9 males, 11 females, 18-79 years old) with unilateral shoulder pathology, using a goniometer and a digital level. Relative reliability was assessed by using intraclass correlation coefficients (ICC), and absolute reliability was assessed by using 95% limits of agreement (LOA). Intratester ICCs ranged from 0.91 to 0.99, and LOA ranged from 3 degrees to 9 degrees for measurements made with the goniometer and digital level. Intertester ICCs ranged from 0.31 to 0.95, and LOA ranged from 6 degrees to 25 degrees . For the comparison of goniometric vs. digital level ROM, ICCs ranged from 0.71 to 0.98. ER and IR ROM were 3-5 degrees greater for the digital level than the goniometer (p < 0.01). Goniometric vs. digital level LOA ranged from 6 degrees to 11 degrees for shoulder flexion. Both measurement techniques had excellent intratester reliability, but for intertester reliability ICCs were 20% lower and LOA were 2.3 times higher than intratester values. Reliability estimates were similar between the digital level and the goniometer. However, because glenohumeral rotation was 3-5 degrees greater for the digital level than the goniometer (systematic error), the two methods cannot be used interchangeably. On the basis of the average intratester LOA for the goniometer and the digital level, a change of 6-11 degrees is needed to be certain that true change has occurred. For comparison of measures made by two different therapists, a change is of 15 degrees is required to be certain a true change has occurred. A digital level can be used to reliably measure shoulder ROM but should not be used interchangeably with a standard goniometer.


A Comprehensive Biophysical Model of Ion and Water Transport in Plant Roots. II. Clarifying the Roles of SOS1 in the Salt-Stress Response in Arabidopsis.
SOS1 transporters play an essential role in plant salt tolerance. Although SOS1 is known to encode a plasma membrane Na+/H+ antiporter, the transport mechanisms by which these transporters contribute to salt tolerance at the level of the whole root are unclear. Gene expression and flux measurements have provided conflicting evidence for the location of SOS1 transporter activity, making it difficult to determine their function. Whether SOS1 transporters load or unload Na+ from the root xylem transpiration stream is also disputed. To address these areas of contention, we applied a mathematical model to answer the question: what is the function of SOS1 transporters in salt-stressed Arabidopsis roots? We used our biophysical model of ion and water transport in a salt-stressed root to simulate a wide range of SOS1 transporter locations in a model Arabidopsis root, providing a level of detail that cannot currently be achieved by experimentation. We compared our simulations with available experimental data to find reasonable parameters for the model and to determine likely locations of SOS1 transporter activity. We found that SOS1 transporters are likely to be operating in at least one tissue of the outer mature root, in the mature stele, and in the epidermis of the root apex. SOS1 transporter activity in the mature outer root cells is essential to maintain low cytosolic Na+ levels in the root and also restricts the uptake of Na+ to the shoot. SOS1 transporters in the stele actively load Na+ into the xylem transpiration stream, enhancing the transport of Na+ and water to the shoot. SOS1 transporters acting in the apex restrict cytosolic Na+ concentrations in the apex but are unable to maintain low cytosolic Na+ levels in the mature root. Our findings suggest that targeted, tissue-specific overexpression or knockout of SOS1 may lead to greater salt tolerance than has been achieved with constitutive gene changes. Tissue-specific changes to the expression of SOS1 could be used to identify the appropriate balance between limiting Na+ uptake to the shoot while maintaining water uptake, potentially leading to enhancements in salt tolerance.


Non-Hodgkin Lymphoma: Examining Mycosis Fungoides and Sézary Syndrome in the Context of Oncology Nursing.
Mycosis fungoides and Sézary syndrome are the most common non-Hodgkin lymphomas that manifest primarily in the skin. Although early-stage disease has an excellent long-term survival rate, advanced disease carries a poor survival rate. Given the lengthy and complex clinical course, nurses are at the forefront of education and supportive care management for patients and caregivers. This article aims to provide an overview of mycosis fungoides and Sézary syndrome and to highlight practice considerations for optimal nursing care. Clinical presentation, diagnosis, management, and nursing consideration are discussed. Oncology nurses have a vital role in educating patients and their caregivers about the side effects of cancer treatment, appropriate skin care, and infection risk.


Tularemia: emergence/re-emergence.
Francisella tularensis is a gram-negative coccobacillus and the etiologic agent of the zoonotic disease tularemia. First described in 1911 in Tulare County, California, it has since been reported throughout the Northern Hemisphere, with natural infections reported among an unusually wide range of vertebrates and invertebrates. In recent years, tularemia has emerged in new geographic locations, populations, and settings. This review will serve to highlight mechanisms contributing to the recent emergence of tularemia as well as a repertoire of diagnostic tools useful for detecting and diagnosing disease.


Aboriginal health.
To inform health care workers about the health status of Canada's native people. A MEDLINE search for articles published from Jan. 1, 1989, to Nov. 31, 1995, with the use of subject headings "Eskimos" and "Indians, North American," excluding specific subject headings related to genetics and history. Case reports were excluded. Material was also identified from a review of standard references and bibliographies and from consultation with experts. Review and research articles containing original data concerning epidemiologic aspects of native health. Studies of Canadian populations were preferred, but population-based studies of US native peoples were included if limited Canadian information was available. Information about target population, methods and conclusions was extracted from each study. Mortality and morbidity rates are higher in the native population than in the general Canadian population. The infant mortality rates averaged for the years 1986 to 1990 were 13.8 per 1000 live births among Indian infants, 16.3 per 1000 among Inuit infants, and only 7.3 per 1000 among all Canadian infants. Age-standardized all-cause mortality rates among residents of reserves averaged for the years 1979 to 1983 were 561.0 per 100,000 population among men and 334.6 per 100,000 among women, compared with 340.2 per 100,000 among all Canadian men and 173.4 per 100,000 among all Canadian women. Compared with the general Canadian population, specific native populations have an increased risk of death from alcoholism, homicide, suicide and pneumonia. Of the aboriginal population of Canada 15 years of age and older, 31% have been informed that they have a chronic health problem. Diabetes mellitus affects 6% of aboriginal adults, compared with 2% of all Canadian adults. Social problems identified by aboriginal people as a concern in their community include substance abuse, suicide, unemployment and family violence. Subgroups of aboriginal people are at a greater-than-normal risk of infectious diseases, injuries, respiratory diseases, nutritional problems (including obesity) and substance abuse. Initial data suggest that, compared with the general population, some subgroups of the native population have a lower incidence of heart disease and certain types of cancer. However, knowledge about contributing factors to the health status of aboriginal people is limited, since the literature generally does not assess confounding factors such as poverty. Canadian aboriginal people die earlier than their fellow Canadians, on average, and sustain a disproportionate share of the burden of physical disease and mental illness. However, few studies have assessed poverty as a confounding factor. Future research priorities in native health are best determined by native people themselves.


Assembly of recombinant Israeli Acute Paralysis Virus capsids.
The dicistrovirus Israeli Acute Paralysis Virus (IAPV) has been implicated in the worldwide decline of honey bees. Studies of IAPV and many other bee viruses in pure culture are restricted by available isolates and permissive cell culture. Here we show that coupling the IAPV major structural precursor protein ORF2 to its cognate 3C-like processing enzyme results in processing of the precursor to the individual structural proteins in a number of insect cell lines following expression by a recombinant baculovirus. The efficiency of expression is influenced by the level of IAPV 3C protein and moderation of its activity is required for optimal expression. The mature IAPV structural proteins assembled into empty capsids that migrated as particles on sucrose velocity gradients and showed typical dicistrovirus like morphology when examined by electron microscopy. Monoclonal antibodies raised to recombinant capsids were configured into a diagnostic test specific for the presence of IAPV. Recombinant capsids for each of the many bee viruses within the picornavirus family may provide virus specific reagents for the on-going investigation of the causes of honeybee loss.


Multiple chemical sensitivity syndrome: symptom prevalence and risk factors in a military population.
To assess the prevalence of and risk factors for self-reported symptoms suggestive of multiple chemical sensitivities/idiopathic environmental intolerance (MCS/IEI) in Persian Gulf War (PGW) veterans from Iowa and a comparison group of PGW-era military personnel. A population-based sample of Iowa military personnel was surveyed using a cross-sectional telephone interview. Study participants were randomly drawn from 1 of 4 domains: PGW active duty, PGW National Guard/Reserve, non-PGW active duty, and non-PGW National Guard/Reserve. A complex sample survey design was used selecting participants from the following substrata: age, sex, race, rank, and military branch. The criteria for MCS/IEI were developed using expert consensus and the medical literature. A total of 3695 study participants (76% of those eligible) completed the telephone survey. The prevalence of symptoms suggestive of MCS/IEI in all participants was 3.4%. Veterans of the PGW reported a significantly higher prevalence of symptoms suggestive of MCS/IEI than did non-PGW military personnel (5.4% vs 2.6%); greater sensitivity to organic chemicals, vehicle exhaust, cosmetics, and smog; and more lifestyle changes. The following risk factors for MCS/IEI were identified with univariate analysis: deployment to the Persian Gulf, age (>25 years), female sex, receiving a physician diagnosis of MCS, previous professional psychiatric treatment, previous psychotropic medication use, current psychiatric illness, and a low level of preparedness. Multiple logistic regression analysis identified several independent risk factors for MCS/IEI, including deployment to the Persian Gulf, age, sex, rank, branch of service, previous professional psychiatric treatment, and current mental illness. Self-reported symptoms suggestive of MCS/IEI are relatively frequent in a military population and are more common among PGW veterans than comparable controls. Reported chemical sensitivities and accompanying behavioral changes were also frequent. After adjusting for age, sex, and training preparedness, previous professional psychiatric treatment and previous psychotropic medication use (before deployment) showed a robust association with symptoms suggestive of MCS.


Cadmium removal by Lemna minor and Spirodela polyrhiza.
The present study investigates the ability of two genus of duckweed (Lemna minor and Spirodela polyrhiza) to phytoremediate cadmium from aqueous solution. Duckweed was exposed to six different cadmium concentrations, such as, 0.5,1.0,1.5, 2.0, 2.5, and 3.0 mg/L and the experiment was continued for 22 days. Water samples were collected periodically for estimation of residual cadmium content in aqueous solution. At the end of treatment period plant samples were collected and accumulated cadmium content was measured. Cadmium toxicity was observed through relative growth factor and changes in chlorophyll content Experimental results showed that Lemna minor and Spirodela polyrhiza were capable of removing 42-78% and 52-75% cadmium from media depending upon initial cadmium concentrations. Cadmium was removed following pseudo second order kinetic model Maximum cadmium accumulation in Lemna minor was 4734.56 mg/kg at 2 mg/L initial cadmium concentration and 7711.00 mg/kg in Spirodela polyrhiza at 3 mg/L initial cadmium concentration at the end of treatment period. Conversely in both cases maximum bioconcentration factor obtained at lowest initial cadmium concentrations, i.e., 0.5 mg/L, were 3295.61 and 4752.00 for Lemna minor and Spirodela polyrhiza respectively. The present study revealed that both Lemna minor and Spirodela polyrhiza was potential cadmium accumulator.


A Double-Edged Sword: The Anti-Cancer Effects of Emodin by Inhibiting the Redox-Protective Protein MTH1 and Augmenting ROS in NSCLC.
Background: Reactive oxygen species (ROS), playing a two-fold role in tumorigenesis, are responsible for tumor formation and progression through the induction of genome instability and pro-oncogenic signaling. The same ROS is toxic to cancer cells at higher levels, oxidizing free nucleotide precursors (dNTPs) as well as damaging DNA leading to cell senescence. Research has highlighted the tumor cell-specific expression of a redox-protective phosphatase, MutT homolog 1 (MTH1), that performs the enzymatic conversion of oxidized nucleotides (like 8-oxo-dGTP) to their corresponding monophosphates, up-regulated in numerous cancers, circumventing their misincorporation into the genomic DNA and preventing damage and cell death. Methods: To identify novel natural small molecular inhibitors of MTH1 to be used as cancer therapeutic agents, molecular screening for MTH1 active site binders was performed from natural small molecular libraries. Emodin was identified as a lead compound for MTH1 active site functional inhibition and its action on MTH1 inhibition was validated on non-small cell lung cancer cellular models (NSCLC). Results: Our study provides strong evidence that emodin mediated MTH1 inhibition impaired NSCLC cell growth, inducing senescence. Emodin treatment enhanced the cellular ROS burdens, on one hand, damaged dNTP pools and inhibited MTH1 function on the other. Our work on emodin indicates that ROS is the key driver of cancer cell-specific increased DNA damage and apoptosis upon MTH1 inhibition. Consequently, we observed a time-dependent increase in NSCL cancer cell susceptibility to oxidative stress with emodin treatment. Conclusions: Based on our data, the anti-cancer effects of emodin as an MTH1 inhibitor have clinical potential as a single agent capable of functioning as a ROS inducer and simultaneous blocker of dNTP pool sanitation in the treatment of NSCL cancers. Collectively, our results have identified for the first time that the potential molecular mechanism of emodin function, increasing DNA damage and apoptosis in cancer cells, is via MTH1 inhibition.


Pharmacists' perceptions and communication of risk for alertness impairing medications.
A core role of the pharmacist is to ensure safe and effective medication use. Therapeutic classes that impair alertness (e.g. sedatives or hypnotics) can pose safety concerns for the consumer when undertaking activities requiring psychomotor vigilance (e.g. driving). To explore pharmacists' perceptions and communication strategy of the risks related to alertness impairing medications in clinical practice. In-depth semi-structured interviews explored community pharmacists' perceptions of medication-related risks, current medication provision and the feasibility of new practice tools. Interviews were digitally recorded, transcribed verbatim and analysed using Framework Analysis to identify emergent themes. A Psychometric Risk Perception Questionnaire was also used to evaluate pharmacists' perceptions across 7 common psychotropic drug classes. Synthesis of the qualitative dataset of 30 pharmacist interviews revealed three key themes: 'Safety and Consequences of AIMs', 'Factors that Influence Risk Communication' and 'Refining Risk Communication'. Participating pharmacists were generally aware of the therapeutic classes associated with medication-related risks but were concerned about patients' level of understanding. Counselling approaches were largely dictated by perceived patient interest/experience with a medication. Concerns were centred on inter-individual pharmacokinetic differences, which could make the precise risk assignment difficult. Pharmacists also highlighted workflow limitations and the need to bring patients' attention to these resources during the clinical interaction to maximise impact. Medication-related risk communication is a complex clinical phenomenon dictated by patients' prior experiences and the pharmacists' practice environment. Extending the evidence base in this therapeutic area and refining clinical resources are key steps towards optimising patient medication safety.


A two-dimensional suspension array system by coupling field flow fractionation to flow cytometry.
Flow field flow fractionation (Fl-FFF) was coupled to flow cytometry to improve the performance of suspension arrays. Size-based separation of the protein-conjugated microspheres by Fl-FFF was performed and the results demonstrated that, the separation could tolerate a wide range of carrier fluid conditions (pH values, salt concentrations, and buffer compositions) favorable for immunoassays. The immuno-complex remained intact during Fl-FFF, as revealed by fluorescence measurements before and after the Fl-FFF separation, and SDS-PAGE of the eluted proteins. The sample throughput of the suspension array can be increased several folds by using particles of different sizes and separating them with Fl-FFF before flow cytometric measurement. Moreover, the gel result hinted that the continuous wash inside the Fl-FFF system may lower the assay background, another possible advantage of the two-dimensional suspension array system.


Human myiasis in patients with diabetic foot: 18 cases.
Myiasis complication of diabetic foot ulcer has only been presented in a few case reports. Therefore, there is a need for additional data on this infestation. Evaluate clinical characteristics of human myiasis in patients with diabetic foot. Case series. A tertiary referral healthcare institution and a diabetic foot center. Patients with diabetic foot infection com.plicated by myiasis who were admitted between June 2012 and July 2017. Bacterial infection rate, accompanying bacterial agents, amputation (morbidity) and mortality rate. 18. Eight (44.4%) of the patients were female. Sixteen (88.9%) had moderate-to-severe infections; 15 (83.3%) had necrotic tissue. Larval debridement therapy was performed on all patients at the bed.side in consecutive sessions. A third-stage larva of Calliphora was detected in one case (5.6%). Second- and third-stage larvae of Lucilia sericata were detected in 5 (27.8%) and 7 (38.9%) patients, respectively. All the patients had a bacterial infection with myiasis. Twelve (66.7%) patients underwent amputation. Three (16.7%) patients died. Myiasis was more frequent in the months of May, June and July. To our knowledge, this is the largest reported series of cases of diabetic foot with myiasis. The most common parasitic agent was Lucilia sericata. Bacterial soft tissue infections were observed in all cases. Poor hygienic conditions were noteworthy and all patients were in need of radical surgery. Myiasis complication of diabetic foot is more frequently seen in the spring and summer. Insufficient follow-up time for analysis of possible confounding factors. None.


Mouse K-Cl cotransporter KCC1: cloning, mapping, pathological expression, and functional regulation.
Although K-Cl cotransporter (KCC1) mRNA is expressed in many tissues, K-Cl cotransport activity has been measured in few cell types, and detection of endogenous KCC1 polypeptide has not yet been reported. We have cloned the mouse erythroid KCC1 (mKCC1) cDNA and its flanking genomic regions and mapped the mKCC1 gene to chromosome 8. Three anti-peptide antibodies raised against recombinant mKCC1 function as immunoblot and immunoprecipitation reagents. The tissue distributions of mKCC1 mRNA and protein are widespread, and mKCC1 RNA is constitutively expressed during erythroid differentiation of ES cells. KCC1 polypeptide or related antigen is present in erythrocytes of multiple species in which K-Cl cotransport activity has been documented. Erythroid KCC1 polypeptide abundance is elevated in proportion to reticulocyte counts in density-fractionated cells, in bleeding-induced reticulocytosis, in mouse models of sickle cell disease and thalassemia, and in the corresponding human disorders. mKCC1-mediated uptake of (86)Rb into Xenopus oocytes requires extracellular Cl(-), is blocked by the diuretic R(+)-[2-n-butyl-6,7-dichloro-2-cyclopentyl-2, 3-dihydro-1-oxo-1H-indenyl-5-yl-)oxy]acetic acid, and exhibits an erythroid pattern of acute regulation, with activation by hypotonic swelling, N-ethylmaleimide, and staurosporine and inhibition by calyculin and okadaic acid. These reagents and findings will expedite studies of KCC1 structure-function relationships and of the pathobiology of KCC1-mediated K-Cl cotransport.


Effect of Bivalent Vaccines against Vibrio anguillarum and Aeromonas salmonicida Subspecie achromogenes on Health and Survival of Turbot.
The efficacy of intraperitoneal injection of an oil-based bivalent autogenous vaccine and the commercial vaccine AlphaJect 3000 (Pharmaq AS) to prevent atypical furunculosis and vibriosis in turbot was analyzed. The effect of both vaccines on health parameters and survival of fish after challenge with V. anguillarum and A. salmonicida subsp. achromogenes was tested. The autogenous vaccine conferred high levels of protection and long-lasting immunity against both pathogens with a single dose. However, severe side effects were observed in turbot injected with this autovaccine and minor negative effects with the AlphaJect 3000 vaccine and the adjuvant Montanide or Eolane. All vaccinated fish showed remarkable antibody agglutination titers, higher than those of control fish, which were maintained 160 d after vaccination. In conclusion, the autogenous bivalent vaccine induces long-lasting protection against atypical furunculosis and vibriosis in turbot, after administration of a single dose, at the cost of high side effects in fish. Therefore, the development of new vaccines should focus on autovaccines and the use of liquid paraffin adjuvants that increase protection with reduced or no side effects.


The effects of streptozotocin-induced diabetes and insulin-treatment on the cardiovascular system of the rat.
In this study the cardiovascular effects of diabetes consisted of a decrease in the heart rate 6 days and in the blood pressure 7 weeks after the induction of streptozotocin-diabetes in rats. The diabetes-induced decrease in heart rate was reversed within 4 days after the institution of insulin-treatment, which also prevented the fall in blood pressure. Maximal KC1 (70 mM) and phenylephrine (10(-4)M)-induced contractures in aortae from diabetic rats were 57 and 48%, respectively, of those from control animals, while tissues from insulin-treated diabetic rats did not differ from controls. Theophylline (10(-2)M)-induced relaxation of the phenylephrine contracture in diabetic tissues was less than in control aortae while relaxation of the K-contracture was greater in control than in diabetic tissues. Insulin-treatment reversed the effects of diabetes on theophylline-induced relaxation of the KC1, but not the phenylephrine contracture. These findings indicate that insulin-treatment will either prevent or reverse diabetes-induced decreases in blood pressure, heart rate and vascular responsiveness to phenylephrine and CK1.


Frequent LOH at chromosome 12q22-23 and Apaf-1 inactivation in glioblastoma.
Glioblastoma (GB) often has loss of heterozygosity on the chromosomes, 1p, 10p, 10q, 11p, 17p, 19q, 22q, and several others. In the case of chromosome 12q, however, it remains to be seen whether LOH occurs. Apaf-1, the apoptotic protease activating factor-1, located at chromosome 12q22-23, is a major effecter of the p53 mediated apoptosis pathway, and Apaf-1 inactivation due to chromosome 12q22-23 LOH and hypermethylation may be involved in some of the neoplasms in malignancy. However, little is known about the frequency of the 12q22-23 LOH or the state of Apaf-1 in GB. To elucidate their involvement in GB, we analyzed a series of 33 GBs for chromosome 12q22-23 LOH, Apaf-1 mRNA expression, and Apaf-1 protein expression, using microsatellite analysis, reverse transcription (RT)-PCR analysis, and immunohistochemical (IHC) analysis, respectively. We also evaluated if and how the 12q22-23 LOH correlated with the p53 gene mutation and EGFR gene amplification. Chromosome 12q22-23 LOH was detected in 14 (42%) of 33 cases. Among the examined cases with LOH at 12q22-23, a low expression of Apaf-1 mRNA was detected in 9 (69%) of 13 cases, and a low expression of Apaf-1 protein was detected in 12 (86%) of 14 cases. The 12q22-23 LOH was significantly correlated with low expression of mRNA and protein (p<0.05, p<0.001 respectively). The p53 gene mutation and EGFR gene amplification were found in 13 cases (39%) and 8 cases (24%), respectively, and these gene alterations were inversely correlated. However, 12q22-23 LOH had no correlations with the p53 gene mutation or EGFR gene amplification. Six of 9 GBs (67%) with neither p53 gene mutation nor EGFR gene amplification tested positive for 12q22-23 LOH. These GBs are likely to belong to another subset independent from the 2 common genetic subsets in GB (one with p53 gene mutation and without EGFR gene amplification, and the other with EGFR gene amplification and without p53 gene mutation). Twenty-three (70%) out of the 33 GBs with the 12q22-23 LOH also tested positive for Apaf-1 inactivation or p53 gene mutation. This high frequency of alterations in the apoptosis-associated factors prompts a speculation that abrogation of the Apaf-1 and p53 mediated apoptosis pathway may play an important role in the tumorigenesis of GB.


Comparison of ceftriaxone and traditional therapy of bacterial meningitis.
Forty-five children (aged 1 day to 15 years) with bacterial meningitis were randomized to receive either traditional therapy (ampicillin and chloramphenicol or gentamicin, pending sensitivity) or ceftriaxone (100 mg/kg per day in two doses for a minimum of 10 days). The etiological agents involved were similar for the two groups and included Haemophilus influenzae type b, Neisseria meningitidis, Streptococcus pneumoniae, and group B streptococcus. Repeat spinal taps were carried out 24 to 48 h after admission. Organisms were seen on the Gram stain of one patient treated with ceftriaxone, but five patients in the traditional therapy group had organisms present on Gram stain of uncentrifuged spinal fluid or positive cultures of the spinal fluid (or both). Ceftriaxone entered the cerebrospinal fluid well, and the average cerebrospinal fluid bactericidal activity for ceftriaxone 1 h after a dose was at least 60 times greater than for ampicillin or chloramphenicol. In those patients who received treatment for a long enough period of time to permit evaluation, there was one death in each group, both due to S. pneumoniae. The length of fever and complications were similar for the patients in both groups. Ceftriaxone was well tolerated; diarrhea, seen in 5 of the 22 patients who received the drug, was the most commonly encountered adverse effect. It was mild, and in no case was it necessary to discontinue the drug. Ceftriaxone appears in this preliminary study to be a safe and acceptable single agent for the treatment of bacterial meningitis in children.


Tumor size is the primary prognosticator for pancreatic cancer after regional pancreatectomy.
The purpose of this study was to evaluate the regional pancreatectomy as surgical therapy for ductal adenocarcinoma of the pancreas and to evaluate potential prognostic factors. Regional pancreatectomy was developed as a more adequate surgical procedure for pancreatic cancer in an attempt to improve the cure rate for this highly lethal disease. Few studies have evaluated large numbers of patients treated with this technique, and in recent years the emphasis has been on more limited surgery for pancreatic cancer. Fifty-six patients with ductal adenocarcinoma of the pancreatic head were treated by regional subtotal or total pancreatectomy. Clinical and pathologic parameters were reviewed and potential prognostic factors were compared statistically. The three patients who died within 30 days of the operation were excluded from the survival analysis. Primary tumor size was the strongest determinant of prognosis. The mean tumor size was 3.9 cm (range, 1-7 cm). Eighty-five percent of patients had peripancreatic soft tissue invasion microscopically, and 58% had regional lymph node metastasis. Kaplan-Meier survival curves indicated a 33% 5-year survival for patients with tumor 2.5 cm or less in diameter (n=12) and 12% for patients with larger tumors (n=39). No patient with a tumor larger than 5 cm survived more than 5 years. Mean tumor size was not significantly associated with lymph node metastases, but 5 of 12 patients (42%) with primary tumor < or =2.5 cm had lymph node metastases. Twenty-four percent of patients with negative lymph nodes and 14% with positive lymph nodes survived 5 years. The difference was not statistically significant (p=0.3), but this is likely related to sample size. The 30- day operative mortality was 5.3%. The most common complications were infection, gastrointestinal bleeding, and gastric stasis. After regional pancreatectomy, tumor size is the strongest predictor of prognosis. A multi- institutional randomized prospective trial of regional pancreatectomy versus pancreaticoduodenectomy is warranted in previously untreated, noninfected cases.


Evaluating the power of surface attendance counts to detect long-term trends in populations of crevice-nesting auklets.
Power analyses are essential when developing a long-term monitoring program for a target species whose observation is logistically challenging and expensive. These analyses can be complicated when the observations have a complex variance structure reflecting many factors. Crevice-nesting seabirds such as least and crested auklets Aethia pusilla and Aethia cristatella illustrate both this need and these challenges. They are ecosystem indicators for the Bering Sea, a system expected to undergo large changes. Unfortunately, they are difficult to monitor as colonies occur on remote, hard to access islands in the Aleutians and Bering Sea, and nests occur in crevices underground, preventing direct observation. Current monitoring consists of breeding-season counts of auklets standing on the surface of sample plots in the colony; logically, a substantial decline in nesting population guarantees an eventual substantial decline in surface attendants. Yet, it remains debatable whether these highly variable counts can be used to statistically detect biologically relevant declines in the attending population let alone the nesting population. Subsequently, existing monitoring programs vary widely in survey design, effort levels, and daily summary statistics. The power of different survey designs was assessed by simulating observations from a state model developed from 11 years of observations using mixed-effects models and zero-inflated Poisson-lognormal regression. The analyses illustrate the process required for any monitoring program whose observations are described inadequately by standard statistical models. State model development revealed survey design refinements that reduce sampling variation. For least auklets, current sampling efforts provided 90% power to detect annual declines of 11% ("Critically Endangered" using IUCN Red List criteria), 4.5% ("Endangered"), or 2.4% ("Vulnerable") in two, four, or six generations, respectively; crested auklets took a few years longer. Power was more sensitive to number of days than number of plots. Results appear robust across a range of bird densities, providing guidance for monitoring other colonies or crevice-nesting species with similar life history strategies. Research should now focus on illuminating the relationship between the attending and nesting populations. Given the frequency of complicated variance structures and zero counts in ecological data, the general statistical models used here should prove widely applicable.


Modulation of growth of human prostate cancer cells by the N-acetylcysteine conjugate of phenethyl isothiocyanate.
There is growing evidence that thiol conjugates of isothiocyanates present in cruciferous vegetables are effective cancer chemopreventive and potentially active therapeutic agents. The effects of the N-acetylcysteine conjugate of phenethyl isothiocyanate (PEITC-NAC) on tumor cell growth were analyzed in human prostate cancer cell lines LNCaP, androgen-dependent, and DU-145, androgen-independent. Exposure of the cells to PEITC-NAC at high concentrations caused cytolysis, while at lower concentrations PEITC-NAC mediated a dose-dependent growth modulation, with reduction of DNA synthesis and growth rate, inhibition of clonogenicity and induction of apoptosis in both types of prostate cancer cells. PEITC-NAC decreased cells in S and G2M phases of cell cycle, blocking cells entering replicating phases. In parallel, a significant enhancement of cells expressing the cell cycle regulator p21 as well as its intensity was determined using a fluorescent antibody technique. The action of PEITC-NAC was time-dependent, with the magnitude of inhibition increasing to 50-65% after PEITC-NAC exposure for several days. Interaction of tumor cells with dissociation products of PEITC-NAC, PEITC and NAC, are proposed as the mechanism of growth regulation.


Modified nucleosides in asbestos workers at high risk of malignant disease: results of a preliminary study applying discriminant analysis.
Patients with asbestos related malignant mesothelioma excrete high levels of modified nucleosides in their urine. The purpose of the present report was to explore further the usefulness of measuring these breakdown products of transfer RNA (tRNA) in male asbestos insulation workers who are at high neoplastic risk but without clinical signs of malignancy. Modified nucleoside levels (psi, m'A, m'I, m2G, and ac4C) were used as discriminator variables in a computer generated discriminant function in which 96% of the controls and 95% of the insulation workers were correctly classified. It was also found, using a similar multiple regression model, that 10 of 13 were correctly classified as having normal chest radiographs and 27 of 30 asbestos exposed subjects as exhibiting alterations in either the parenchyma, pleura, or both. The results suggest that measuring modified nucleosides levels in the urine of asbestos exposed workers, and perhaps others exposed to carcinogenic agents, has the potential for identifying, through multivariate statistical techniques, individuals who are at high neoplastic risk.


MRI of tumor T cell infiltration in response to checkpoint inhibitor therapy.
Immune checkpoint inhibitors, the most widespread class of immunotherapies, have demonstrated unique response patterns that are not always adequately captured by traditional response criteria such as the Response Evaluation Criteria in Solid Tumors or even immune-specific response criteria. These response metrics rely on monitoring tumor growth, but an increase in tumor size and/or appearance after starting immunotherapy does not always represent tumor progression, but also can be a result of T cell infiltration and thus positive treatment response. Therefore, non-invasive and longitudinal monitoring of T cell infiltration are needed to assess the effects of immunotherapies such as checkpoint inhibitors. Here, we proposed an innovative concept that a sufficiently large influx of tumor infiltrating T cells, which have a smaller diameter than cancer cells, will change the diameter distribution and decrease the average size of cells within a volume to a degree that can be quantified by non-invasive MRI. We validated our hypothesis by studying tumor response to combination immune-checkpoint blockade (ICB) of anti-PD-1 and anti-CTLA4 in a mouse model of colon adenocarcinoma (MC38). The response was monitored longitudinally using Imaging Microstructural Parameters Using Limited Spectrally Edited Diffusion (IMPULSED), a diffusion MRI-based method which has been previously shown to non-invasively map changes in intracellular structure and cell sizes with the spatial resolution of MRI, in cell cultures and in animal models. Tumors were collected for immunohistochemical and flow cytometry analyzes immediately after the last imaging session. Immunohistochemical analysis revealed that increased T cell infiltration of the tumors results in a decrease in mean cell size (eg, a 10% increase of CD3+ T cell fraction results a ~1 µm decrease in the mean cell size). IMPULSED showed that the ICB responders, mice with tumor volumes were less than 250 mm3 or had tumors with stable or decreased volumes, had significantly smaller mean cell sizes than both Control IgG-treated tumors and ICB non-responder tumors. IMPULSED-derived cell size could potentially serve as an imaging marker for differentiating responsive and non-responsive tumors after checkpoint inhibitor therapies, a current clinical challenge that is not solved by simply monitoring tumor growth.


Sand-induced diarrhea in a foal.
A diagnosis of sand enteropathy was made in a 3 1/2-month-old Quarter Horse filly. Clinical signs included diarrhea and weight loss of 2 1/2 months' duration. Abdominal radiographs were useful in diagnosis of the condition and in evaluating response to therapy. The filly responded to treatment with psyllium hydrophilic mucilloid although full recovery took 2 months. Diagnosis and treatment of sand enteropathy is discussed.


Quantitative Analysis of Phenolic Acids and Antiplatelet Activity of Melissa officinalis Leaf Extracts.
The contents of total phenolic acids and rosmarinic acid were determined and the antiplatelet potential assessed of extracts of lemon balm (Melissa officinalis L., Lamiaceae) leaf samples collected during 2014 and 2015. The quantity of total phenolic acids ranged from 4.8 to 8.5%, while the rosmarinic acid content ranged from 1.78 to 4.35%. Results of statistical analysis showed that the undiluted extracts have antiplatelet activity in the ADP test (final concentration 31.25 mg/L). While there are reports on antiplatelet activity of lemon balm essential oil, to the best of our knowledge, this is the first report on antiplatelet activity of lemon balm leaf extracts.


Comparison of Poisson and Bernoulli spatial cluster analyses of pediatric injuries in a fire district.
With limited resources available, injury prevention efforts need to be targeted both geographically and to specific populations. As part of a pediatric injury prevention project, data was obtained on all pediatric medical and injury incidents in a fire district to evaluate geographical clustering of pediatric injuries. This will be the first step in attempting to prevent these injuries with specific interventions depending on locations and mechanisms. There were a total of 4803 incidents involving patients less than 15 years of age that the fire district responded to during 2001-2005 of which 1997 were categorized as injuries and 2806 as medical calls. The two cohorts (injured versus medical) differed in age distribution (7.7 +/- 4.4 years versus 5.4 +/- 4.8 years, p < 0.001) and location type of incident (school or church 12% versus 15%, multifamily residence 22% versus 13%, single family residence 51% versus 28%, sport, park or recreational facility 3% versus 8%, public building 8% versus 7%, and street or road 3% versus 30%, respectively, p < 0.001). Using the medical incident locations as controls, there was no significant clustering for environmental or assault injuries using the Bernoulli method while there were four significant clusters for all injury mechanisms combined, 13 clusters for motor vehicle collisions, one for falls, and two for pedestrian or bicycle injuries. Using the Poisson cluster method on incidence rates by census tract identified four clusters for all injuries, three for motor vehicle collisions, four for fall injuries, and one each for environmental and assault injuries. The two detection methods shared a minority of overlapping geographical clusters. Significant clustering occurs overall for all injury mechanisms combined and for each mechanism depending on the cluster detection method used. There was some overlap in geographic clusters identified by both methods. The Bernoulli method allows more focused cluster mapping and evaluation since it directly uses location data. Once clusters are found, interventions can be targeted to specific geographic locations, location types, ages of victims, and mechanisms of injury.


Health status of people of slums in Nairobi, Kenya.
The objective of this study was to describe the health status of people living in the slums of Nairobi. It was designed as a cross-sectional study based on data from visitors at a clinic at Trnava University located in Mukuru slum in Nairobi. There were about 16,000 visits registered at Mary Immaculate Clinic of Trnava University in Nairobi during 2 years of operation. A random 5% sample was drawn from the paper-card database of this clinic to assess basic characteristics and health complaints of visitors. Both self-reported health complaints and diagnoses written by physicians were used to assess health status of participants. More females with average age (by slum) ranging from 20.46 to 21.30 years than males with average age ranging from (by slum) 15.86 to 19.49 years are the visitors of the clinic. The major self-reported health complaints of visitors were cough, abdominal pain, and headache for both sexes. The most frequent diagnoses were consequently virosis, acute respiratory infections, and bronchitis. Differences in health complaints by slums were observed and are described herein. The major health complaints and diagnoses in addition to the differences in health complaints and diagnoses by slum show that environmental conditions can have major influences on health status. Therefore, environmental improvements are important in the improvement of health status. A very high prevalence of respiratory complaints and gastrointestinal problems signify that improvements in air pollution reduction, drinking water provision, and waste management in slums can lead to more significant and sustainable improvements in health status than just simple treatment. This fact should be taken into account when planning future relief programs.


Revision Total Elbow Arthroplasty.
Despite recent technologic advances, total elbow arthroplasty has complication rates higher than that of total joint arthroplasty in other joints. With new antirheumatic treatments, the population receiving total elbow arthroplasty has shifted from patients with rheumatoid arthritis to those with posttraumatic arthritis, further compounding the high complication rate. The most common reasons for revision include infection, aseptic loosening, fracture, and component failure. Common mechanisms of total elbow arthroplasty failure include infection, aseptic loosening, fracture, component failure, and instability. Tension band fixation, allograft struts with cerclage wire, and/or plate and screw constructs can be used for fracture stabilization.


Lentiviral-mediated RNAi knockdown yields a novel mouse model for studying Cyp2b function.
There are few in vivo knockout models available to study the function of Cyp2 members involved in the metabolism of endogenous and exogenous chemicals. These models may help provide insight into the cytochrome P450s (CYPs) responsible for the detoxification and activation of drugs, environmental toxicants, and endobiotics. The aim of this work is to produce a potent Cyp2b-knockdown (KD) mouse for subsequent study of Cyp2b function. We made a quintuple Cyp2b-KD mouse using lentiviral-promoted short hairpin RNA (shRNA) homologous to all five murine Cyp2b subfamily members (Cyp2b9, 2b10, 2b13, 2b19, and 2b23). The Cyp2b-KD mice are viable, fertile, and without obvious gross abnormalities except for an increase in liver weight. Expression of the three hepatic Cyp2b members, 2b9, 2b10, and 2b13, is significantly repressed as demonstrated by quantitative real-time PCR and Western blotting. The constitutive androstane receptor activator, 1,4-Bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP), was used to determine if shRNA-mediated Cyp2b10 repression could be outcompeted by Cyp2b10 induction. TCPOBOP-treated Cyp2b-KD mice show 80-90% less Cyp2b protein expression than TCPOBOP-treated wild-type (WT) mice, demonstrating that Cyp induction does not outcompete the repressive function of the shRNA. Untreated and TCPOBOP-treated Cyp2b-KD mice are poor metabolizers of parathion compared with WT mice. Furthermore, Cyp2b-KD mice are sensitive to parathion, an organophosphate insecticide primarily metabolized by Cyp2b enzymes, when compared with WT mice. In summary, we designed an shRNA construct that repressed the expression and activity of multiple Cyp2b enzymes. We foresee that this novel Cyp2b-KD mouse model will significantly improve our understanding of the role of Cyp2b enzymes in chemical sensitivity and drug metabolism.


Distinct domains in the ARC region of the potato resistance protein Rx mediate LRR binding and inhibition of activation.
Plant nucleotide binding and leucine-rich repeat (NB-LRR) proteins contain a region of homology known as the ARC domain located between the NB and LRR domains. Structural modeling suggests that the ARC region can be subdivided into ARC1 and ARC2 domains. We have used the potato (Solanum tuberosum) Rx protein, which confers resistance to Potato virus X (PVX), to investigate the function of the ARC region. We demonstrate that the ARC1 domain is required for binding of the Rx N terminus to the LRR domain. Domain-swap experiments with Rx and a homologous disease resistance gene, Gpa2, showed that PVX recognition localized to the C-terminal half of the LRR domain. However, inappropriate pairings of LRR and ARC2 domains resulted in autoactive molecules. Thus, the ARC2 domain is required to condition an autoinhibited state in the absence of elicitor as well as for the subsequent elicitor-induced activation. Our data suggest that the ARC region, through its interaction with the LRR, translates elicitor-induced modulations of the C terminus into a signal initiation event. Furthermore, we demonstrate that physical disruption of the LRR-ARC interaction is not required for signal initiation. We propose instead that this activity can lead to multiple rounds of elicitor recognition, providing a means of signal amplification.


The highly conserved N-terminal domains of histones H3 and H4 are required for normal cell cycle progression.
The N-terminal domains of the histones H3 and H4 are highly conserved throughout evolution. Mutant alleles deleted for these N-terminal domains were constructed in vitro and examined for function in vivo in Saccharomyces cerevisiae. Cells containing a single deletion allele of either histone H3 or histone H4 were viable. Deletion of the N-terminal domain of histone H4 caused cells to become sterile and temperature sensitive for growth. The normal cell cycle progression of these cells was also altered, as revealed by a major delay in progression through the G2 + M periods. Deletion of the N-terminal domain of histone H3 had only minor effects on mating and the temperature-sensitive growth of mutant cells. However, like the H4 mutant, the H3 mutants had a significant delay in completing the G2 + M periods of the division cycle. Double mutants containing N-terminal domain deletions of both histone H3 and histone H4 were inviable. The phenotypes of cells subject to this synthetic lethality suggest that the N-terminal domains are required for functions essential throughout the cell division cycle and provide genetic evidence that histones are randomly distributed during chromosome replication.


Discharges Against Medical Advice at a County Hospital: Provider Perceptions and Practice.
Patients discharged against medical advice (AMA) have higher rates of readmission and mortality than patients who are conventionally discharged. Bioethicists have proposed best practice approaches for AMA discharges, but studies have revealed that some providers have misconceptions about their roles in these discharges. This study assessed patient characteristics and provider practices for AMA discharges at a county hospital and provider perceptions and knowledge about AMA discharges. This mixed-methods cross-sectional study involved chart abstraction and survey administration. Charts were reviewed for all AMA discharges (n = 319) at a county hospital in 2014. Surveys were completed by 178 healthcare providers at the hospital. Of 12,036 admissions, 319 (2.7%) ended with an AMA discharge. Compared with conventionally discharged patients, patients who left AMA were more likely to be young, male, and homeless and less likely to be Spanish-speaking. Of the AMA patients, 29.6% had capacity documented, 21.4% had medications prescribed, and 25.7% had follow-up arranged. Of patients readmitted within 6 months after AMA, 23.5% left AMA again at the next visit. Attending physicians and trainee physicians were more likely than nurses to say that AMA patients should receive medications and follow-up (94% and 84% vs 64%; P < 0.05). Although providers overall felt comfortable determining capacity and discussing AMA discharges, they rarely documented these discussions. Nurses and physicians differed in their thinking regarding whether to arrange follow-up for patients leaving AMA, and in practice arrangements were seldom made. Journal of Hospital Medicine 2017;12:11-17.


[Effects of climatic treatment in the Bairam-Ali sanatorium on the course of chronic glomerulonephritis].
A study was made of the survival, frequency and time of the development of chronic renal failure (CRF) in 712 patients with chronic glomerulonephritis (CGN) treated more than once (2 to 23 times) in the sanatorium Bairam-Ali. It was shown that a more benign course was typical of latent and hypertonic forms among the clinical ones and mesangioproliferative, mesangiomembranous and membranous variants among the morphological variants of CRF. It should be noted that CRF frequency in these groups was lower and developed much later than in the other CGN variants. Prognosis was unfavorable in the patients over 50. The highest survival was noted among the patients aged 20 to 29. Tonsillectomy did not make a positive effect on the CGN course.


AtnMat2, a nuclear-encoded maturase required for splicing of group-II introns in Arabidopsis mitochondria.
Mitochondria (mt) in plants house about 20 group-II introns, which lie within protein-coding genes required in both organellar genome expression and respiration activities. While in nonplant systems the splicing of group-II introns is mediated by proteins encoded within the introns themselves (known as "maturases"), only a single maturase ORF (matR) has retained in the mitochondrial genomes in plants; however, its putative role(s) in the splicing of organellar introns is yet to be established. Clues to other proteins are scarce, but these are likely encoded within the nucleus as there are no obvious candidates among the remaining ORFs within the mtDNA. Intriguingly, higher plants genomes contain four maturase-related genes, which exist in the nucleus as self-standing ORFs, out of the context of their evolutionary-related group-II introns "hosts." These are all predicted to reside within mitochondria and may therefore act "in-trans" in the splicing of organellar-encoded introns. Here, we analyzed the intracellular locations of the four nuclear-encoded maturases in Arabidopsis and established the roles of one of these genes, At5g46920 (AtnMat2), in the splicing of several mitochondrial introns, including the single intron within cox2, nad1 intron2, and nad7 intron2.


Dielectrophoretic force-induced wrinkling of graphene oxide: Enhancing electrical conductivity and expanding biosensing applications.
Graphene oxide (GO) has many advantages, making it suitable for various applications. However, it has low electrical conductivity, restricting its applicability to electrochemical biosensors. This study used dielectrophoretic (DEP) force to control the movement and deformation of GO nanosheets to achieve high electrical conductivity without the chemical reduction of oxygen functional groups. Subjecting the DEP force to GO nanosheets induced physical deformation leading to the formation of wrinkled structures. A computational simulation was performed to set an appropriate electrical condition for operating a positive DEP force effect of at least 1019 v2/m3, and the interdigitated microelectrode structure was selected. The resulting wrinkled GO exhibited significantly improved electrical conductivity, reaching 21.721 μS while preserving the essential oxygen functional groups. Furthermore, a biosensor was fabricated using wrinkled GO deposited via DEP force. The biosensor demonstrated superior sensitivity, exhibiting a 9.6-fold enhancement compared with reduced GO (rGO) biosensors, as demonstrated through biological experiments targeting inducible nitric oxide synthase. This study highlights the potential of using DEP force to enhance electrical conductivity in GO-based biosensing applications, opening new avenues for high-performance diagnostics.


Urine pentamidine as an indicator of lung pentamidine in patients receiving aerosol therapy.
To determine if urine pentamidine was reflective of lung pentamidine, we compared levels of the drug in bronchoalveolar lavage fluid and simultaneously obtained urine. Thirty-one patients who were receiving aerosolized pentamidine either as treatment or as prophylaxis underwent BAL and submitted urine samples for pentamidine analysis. Pentamidine was analyzed in both phases of BAL fluid (supernatant and cell pellet) and in urine using high performance liquid chromatography. Urine results were normalized for creatinine. Patients were categorized as prophylaxis failures (active Pneumocystis carinii pneumonia on prophylaxis), electives (free from PCP on prophylaxis), treatment (daily AP in treatment doses for active PCP, or miscellaneous (single dose of AP). Levels in BAL fluid and urine varied widely over several orders of magnitude. However, for all patients, we found a highly significant relationship between BAL supernatant and urine (r = 0.97, p less than 0.0001). No statistical differences were found when comparing levels of pentamidine between failures and electives; however, the number of failures was small. We conclude that urine pentamidine is related to lung pentamidine and can be used as a clinical indicator in patients receiving aerosolized therapy.


NF-κB is required for Smac mimetic-mediated sensitization of glioblastoma cells for γ-irradiation-induced apoptosis.
Evasion of apoptosis contributes to radioresistance of glioblastoma, calling for novel strategies to overcome apoptosis resistance. In this study, we investigated the potential of the small molecule Smac mimetic BV6 to modulate radiosensitivity of glioblastoma cells. Here, we identify a novel proapoptotic function of NF-κB in γ-irradiation-induced apoptosis of glioblastoma cells by showing, for the first time, that NF-κB is critically required for Smac mimetic-mediated radiosensitization. BV6 significantly increases γ-irradiation-triggered apoptosis in several glioblastoma cell lines in a dose- and time-dependent manner. Calculation of combination index (CI) reveals that the interaction of BV6 and γ-irradiation is highly synergistic (CI < 0.3). Molecular studies show that BV6 stimulates NF-κB activation, which is critical for radiosensitization, because genetic inhibition of NF-κB by overexpression of the dominant-negative superrepressor IκBα-SR significantly decreases BV6- and γ-irradiation-induced apoptosis. Also, the BV6-mediated enhancement of γ-irradiation-triggered caspase activation, drop of mitochondrial membrane potential, and cytochrome c release is abolished in cells overexpressing IκBα-SR. Similarly, NF-κB inhibition by ectopic expression of a kinase dead mutant of IKKβ prevents the BV6-mediated sensitization for γ-irradiation. The clinical relevance is underscored by experiments with primary tumor samples showing that BV6 sensitizes primary cultured glioma cells as well as glioblastoma-initiating cancer stem cells derived from surgical specimens for γ-irradiation. In conclusion, we identify NF-κB as a critical mediator of Smac mimetic-conferred radiosensitization of glioblastoma cells. These results have important implications for the development of Smac mimetic-based combination protocols for radiosensitization of glioblastoma.


Adolescent food insecurity in female rodents and susceptibility to diet-induced obesity.
Food insecurity is defined as having limited or uncertain access to nutritious foods, and adolescent food insecurity is associated with obesity and disordered eating behaviors in humans. We developed a rodent model of adolescent food insecurity to determine whether adolescent food insecurity per se promotes increased susceptibility to diet-induced obesity and altered eating behaviors during adulthood. Female juvenile Wistar rats were singly housed and assigned to three experimental diets: food-secure with standard chow (CHOW), food-secure with a high-fat/sugar Western diet (WD), and food-insecure with WD (WD-FI). Food-secure rats (CHOW and WD) received meals at fixed feeding times (9:00, 13:00, and 16:00). WD-FI rats received meals at unpredictable intervals of the above-mentioned feeding times but had isocaloric amounts of food to WD. We investigated the impact of adolescent food insecurity on motivation for sucrose (Progressive Ratio), approach-avoidance behavior for palatable high-fat food (Approach-Avoidance task), and susceptibility to weight gain and hyperphagia when given an obesogenic choice diet. Secondary outcomes were the effects of food insecurity during development on anxiety-like behaviors (Open Field and Elevated Plus Maze) and learning and memory function (Novel Location Recognition task). Rodents with adolescent food insecurity showed a greater trend of weight gain and significantly increased fat mass and liver fat accumulation on an obesogenic diet in adulthood, despite no increases in motivation for sucrose or high-fat food. These data suggest that adolescent unpredictable food access increases susceptibility to diet-induced fat gain without impacting food motivation or food intake in female rodents. These findings are among a small group of recent studies modeling food insecurity in rodents and suggest that adolescent food insecurity in females may have long-term implications for metabolic physiology later in life.


Staff perceptions of change resulting from participation in a European cancer accreditation programme: a snapshot from eight cancer centres.
Healthcare accreditation is considered to be an essential quality improvement tool. However, its effectiveness has been critiqued. Twenty-four interviews were conducted with clinicians (five), nurses (six), managers (eight), and basic/translational researchers (five) from eight European cancer centres on changes observed from participating in a European cancer accreditation programme. Data were thematically analysed and verified with participants and checked against auditor's feedback. Four change categories emerged: (i) the growing importance of the nursing and supportive care field (role change). Nurses gained more autonomy/clarity on their daily duties. Importance was given to the hiring and training of supportive care personnel (ii) critical thinking on data integration (strategic change). Managers gained insight on how to integrate institutional level data (iii) improved processes within multidisciplinary team (MDT) meetings (procedural change). Clinical staff experienced improved communication between MDTs (iv) building trust (organisational change). Accreditation improved the centre's credibility with its own staff and externally with funders and patients. No motivational changes were perceived. Researchers perceived no changes. The auditor's feedback included changes in 13 areas: translational research, biobanks, clinical trials, patient privacy and satisfaction, cancer registries, clinical practice guidelines, patient education, screening, primary prevention, role of nurses, MDT, supportive care, and data integration. However, our study revealed that staff perceived changes only in the last four areas. Staff perceived changes in data integration, nursing and supportive care, and in certain clinical aspects. Accreditation programmes must pay attention to the needs of different stakeholder groups, track changes, and observe how/why change happens.


Discrimination between the tastes of sucrose and monosodium glutamate in rats.
Conditioned taste aversion studies have demonstrated that rats conditioned to avoid monosodium glutamate (MSG) with amiloride added to reduce the intensity of the sodium component of MSG taste, will generalize an aversion for MSG to sucrose and vice versa. This suggests that taste transduction for sodium, sucrose and MSG may intersect at some point. Generalization of conditioned taste aversion indicates that two substances share similar taste features, but it does not reveal the extent of their differences. In this study, we tested how well rats can discriminate sucrose and MSG under a variety of conditions. Water-deprived rats were trained on a combination of water reinforcement and shock avoidance to discriminate between MSG and sucrose, both with and without amiloride, and with and without equimolar NaCl in all solutions. In the absence of amiloride, rats reliably distinguished between MSG and sucrose down to 10 mM solutions. However, they could correctly identify solutions only above 50 mM in the presence of amiloride, equimolar sodium chloride, or both. These results suggest that gustatory stimulation by MSG and sucrose interact somewhere in taste transduction, perhaps within taste receptor cells or gustatory afferent pathways.