Subjective judgements of quality of life: a comparison study between people with intellectual disability and those without disability.
The present study investigated the relationship between objective and subjective measures of quality of life (QoL), and in particular, health status and primary care services. Thirty-one people with intellectual disability (ID) were interviewed using a QoL questionnaire. Thirty-one matched controls from the general population filled in a parallel questionnaire. The results in both groups support the findings of many researchers which indicate that satisfaction with aspects of life is generally high (i.e. approximate 75% of maximum possible satisfaction). The present study also confirms previous findings which indicate that satisfaction with aspects of life does not generally correlate with objective circumstances in either group. The importance that people attach to aspects of their lives did not generally correlate with their objective circumstances or their satisfaction with life. However, people with ID attached greater importance to all aspects of their lives than those without disability. This may be linked to their aspirations, preferences and opportunities for choice, and therefore, these factors may be a more meaningful way of considering their QoL. The present findings call into question the use of satisfaction as a general measure of QoL, and also indicate that the use of importance as a mediating variable in understanding satisfaction may not be a solution. The participants with ID had poorer health than the controls, and were significantly less satisfied with their health. Contrary to other findings, the present participants with ID reported that they had received more health checks than the controls over the previous year. Half of these checks had been carried out by 'special' services rather than primary care services.


Cost-effectiveness of adjuvant radiotherapy in intermediate risk endometrial cancer.
Endometrial cancer is the most common gynecologic malignancy in the United States. Adjuvant radiotherapy in patients with intermediate risk disease (stage IB, IC, and occult stage II) is controversial. Despite no proven survival advantage, a significant number of women undergo this treatment annually. The purpose of this study was to compare the estimated health and economic outcomes for adjuvant whole pelvic radiotherapy to no treatment with salvage therapy for recurrence. A decision analytic model was created to estimate the costs of adjuvant pelvic radiotherapy versus no adjuvant radiotherapy in patients with intermediate risk endometrial cancer. Data used was gathered from published literature and institutional data on costs. The model incorporates complications, recurrence rates, treatment of recurrence, and survival in each group. In the base case analysis, adjuvant pelvic radiation reduced the recurrence rate by 50%. Cost-effectiveness as measured by cost per recurrence prevented was highly sensitive to the probability of recurrence and the efficacy of adjuvant therapy. In our model the mean costs of Strategy 1 with observation and treatment reserved until the time of recurrence would be $5016. In contrast the mean cost of Strategy 2 which incorporated adjuvant radiotherapy would be $21,159. Cost per recurrence prevented based on the incremental cost-effectiveness is thus $225,215. In the highest risk subgroup, using the upper limit of the 90% confidence limit of efficacy seen in GOG Protocol 99, cost/recurrence prevented was approximately $50,000. Results did not differ when using parameters solely from GOG 99 or PORTEC. Although adjuvant pelvic radiation does not appear to improve survival for intermediate risk endometrial cancer patients, it does prevent recurrences, at a net positive cost compared to no therapy. Data are not currently available to incorporate quality of life information into cost-effectiveness analyses. Obtaining such data would allow cost/quality-adjusted life year gained to be estimated. This information is necessary to determine if the extra costs of adjuvant radiotherapy in patients with intermediate risk endometrial cancer are acceptable by current health care policy standards.


Thermal hydrolysis of sewage sludge: Improvement in biogas generation and prediction of global warming potential.
Anaerobic digestion (AD) is a prominent treatment method for the sludge produced from sewage treatment plants. Poor solid reduction and longer retention time are the main drawbacks of AD. Thermal hydrolysis (TH) is a potential pretreatment method for solubilization of sewage sludge (SS) solids thereby improving biogas production during AD post-treatment. In this study, the SS sample (total solids = 1.75 wt% and total chemical oxygen demand (COD) = 15,450 mg L-1) was subjected to TH pretreatment (temperature = 140-180°C and reaction time = 60 minutes) in a 0.7-L capacity stainless-steel high-pressure reactor. At a reaction temperature of 180°C, the maximum solid solubilization (total dissolved solids = 4652 mg L-1) and improved dewaterability (time to filter = 4.7 s.L g-1) were observed. The biochemical methane potential test results showed almost doubling of methane generation from 145 to 284 mL gCOD-1 after TH pretreatment at 180°C. The life cycle assessment approach was used to compare various SS treatment and disposal scenarios, two of which included hydrothermal pretreatment. The scenarios involving hydrothermal pretreatments showed the least global warming potential.


Two stage reconstruction for the Shepherd's crook deformity in a case of polyostotic fibrous dysplasia.
Polyostotic fibrous dysplasia leads to progressive and disabling deformity involving the proximal femur. Conventional methods of treatment have been ineffective in controlling this problem. Two stage reconstruction was carried out in a case of polyostotic fibrous dysplasia with bilateral shepherd's crook deformity. Bilateral subtrochanteric osteotomies with intramedullary fixation in the first stage and intertrochanteric osteotomies in the second stage with nail plate fixation was done to provide definitive control of the deformity. Bone graft was not used.


Factors affecting the urinary excretion of endogenously formed dimethyltryptamine in normal human subjects.
The hallucinogenic substance N',N'-dimethyltryptamine and its precursor N-methyltryptamine were found in 24-h specimens of urine from 19 normal human subjects; the mean excretion rates were 386 ng 24 h(-1) and 856 ng 24 h(-1) respectively. The urinary excretion of both compounds was unrelated to age, sex, urinary volume, or creatinine, nor was any consistent diurnal pattern observed. Rates for the mono and dimethylated compounds were not correlated. Diet and the intestinal flora were excluded as a source of urinary dimethyltryptamine. Administration to 4 subjects of sufficient ammonium chloride to increase the H ion concentration of the urine caused a transient increase in dimethyltryptamine excretion but no consistent increase in the rate for N-methyltryptamine. Acidification of the urine did not appear to be the determining factor in this result since in one subject the same drop in urinary pH was achieved by feeding methionine without any increase in dimethyltryptamine excretion.


Biological uptake of polychlorinated biphenyls by Macoma balthica from sediment amended with activated carbon.
This work characterizes the efficacy of activated carbon amendment in reducing polychlorinated biphenyl (PCB) bioavailability to clams (Macoma balthica) from field-contaminated sediment (Hunters Point Naval Shipyard, San Francisco Bay, CA, USA). Test methods were developed for the use of clams to investigate the effects of sediment amendment on biological uptake. Sediment was mixed with activated carbon for one month. Bioaccumulation tests (28 d) were employed to assess the relationships between carbon dose and carbon particle size on observed reductions in clam biological uptake of PCBs. Extraction and cleanup protocols were developed for the clam tissue. Efficacy of activated carbon treatment was found to increase with both increasing carbon dose and decreasing carbon particle size. Average reductions in bioaccumulation of 22, 64, and 84% relative to untreated Hunters Point sediment were observed for carbon amendments of 0.34, 1.7, and 3.4%, respectively. Average bioaccumulation reductions of 41, 73, and 89% were observed for amendments (dose = 1.7% dry wt) with carbon particles of 180 to 250, 75 to 180, and 25 to 75 microm, respectively, in diameter, indicating kinetic phenomena in these tests. Additionally, a biodynamic model quantifying clam PCB uptake from water and sediment as well as loss through elimination provided a good fit of experimental data. Model predictions suggest that the sediment ingestion route contributed 80 to 95% of the PCB burdens in the clams.


Comparison of zinc complexation properties of dissolved organic matter from surface waters and wastewater treatment plant effluents.
Unlike natural organic matter(NOM), wastewater organic matter(WWOM) from wastewater treatment plant effluents has not been extensively studied with respect to complexation reactions with heavy metals such as copper or zinc. In this study, organic matter from surface waters and a wastewater treatment plant effluent were concentrated by reverse osmosis (RO) method. The samples were treated in the laboratory to remove trace metals and major cations. The zinc complexing properties of both NOM and the WWOM were studied by square wave anodic stripping voltammetry(SWASV). Experimental data were compared to predictions using the Windermere Humic Aqueous Model (WHAM) Version VI. We found that the zinc binding of WWOM was much stronger than that of NOM and not well predicted by WHAM. This suggests that in natural water bodies that receive wastewater treatment plant effluents the ratio of WWOM to NOM must be taken into account in order to accurately predict free zinc activities.


Human liver-kidney model elucidates the mechanisms of aristolochic acid nephrotoxicity.
Environmental exposures pose a significant threat to human health. However, it is often difficult to study toxicological mechanisms in human subjects due to ethical concerns. Plant-derived aristolochic acids are among the most potent nephrotoxins and carcinogens discovered to date, yet the mechanism of bioactivation in humans remains poorly understood. Microphysiological systems (organs-on-chips) provide an approach to examining the complex, species-specific toxicological effects of pharmaceutical and environmental chemicals using human cells. We microfluidically linked a kidney-on-a-chip with a liver-on-a-chip to determine the mechanisms of bioactivation and transport of aristolochic acid I (AA-I), an established nephrotoxin and human carcinogen. We demonstrate that human hepatocyte-specific metabolism of AA-I substantially increases its cytotoxicity toward human kidney proximal tubular epithelial cells, including formation of aristolactam adducts and release of kidney injury biomarkers. Hepatic biotransformation of AA-I to a nephrotoxic metabolite involves nitroreduction, followed by sulfate conjugation. Here, we identify, in a human tissue-based system, that the sulfate conjugate of the hepatic NQO1-generated aristolactam product of AA-I (AL-I-NOSO3) is the nephrotoxic form of AA-I. This conjugate can be transported out of liver via MRP membrane transporters and then actively transported into kidney tissue via one or more organic anionic membrane transporters. This integrated microphysiological system provides an ex vivo approach for investigating organ-organ interactions, whereby the metabolism of a drug or other xenobiotic by one tissue may influence its toxicity toward another, and represents an experimental approach for studying chemical toxicity related to environmental and other toxic exposures.


Patients With Cancer and COVID-19: A WhatsApp Messenger-Based Survey of Patients' Queries, Needs, Fears, and Actions Taken.
This descriptive investigation was undertaken at three oncology units to report queries, needs, and fears related to severe acute respiratory syndrome coronavirus 2 (COVID-19) of patients with cancer and to avoid uncontrolled treatment delays or withdrawal, behavioral mistakes, and panic. All queries spontaneously delivered through the WhatsApp instant messaging system commonly used by patients to communicate with oncology units were collected and grouped by homology in five categories. Responses to the queries were given according to recommendations by the Italian Association of Medical Oncology through WhatsApp and by subsequent phone calls. Patients were also classified according to the site of the primary tumor, stage of disease, and current treatments. Analysis of the association between these data and queries was carried out. The social scenario in Italy is a nationwide lockdown except for hospitals, pharmacies, and food supplies. Overall, 446 different patients' WhatsApp conversations were analyzed between March 1 and March 13 and comprised the following: requirement of visit delay by patients undergoing oral therapies or in follow-up, delays in chemotherapy or immunotherapy administration, queries about possible immunosuppression, and changes in lifestyle or daily activities. Delay requirements were statistically more frequent among patients with prostate or breast cancer compared with those with lung or pancreatic cancer. Actions taken by oncologists are also reported. To our knowledge, the WhatsApp instant messaging system has been occasionally used in other medical settings with controversial results. In our experience, WhatsApp turned out to be adequate to give a rapid answer to most queries from patients with cancer in the COVID-19 pandemic scenario.


N-(2-morpholin-4-yl-ethyl)-2-(1naphthyloxy)acetamide inhibits the chronic constriction injury-generated hyperalgesia via the antagonism of sigma-1 receptors.
The most used therapeutic treatment to relieve neuropathic pain is that of neuromodulators such as anti-epileptics or anti-depressants; however, there are alternatives that may be potentially useful. The sigma-1 receptor is a therapeutic target that has shown favorable results at preclinical levels. The aim of this study was to evaluate the anti-hyperalgesic effect of N-(2-morpholin-4-yl-ethyl)-2-(1-naphthyloxy) acetamide (NMIN) in a chronic constriction injury model (CCI) and compare it both a sigma-1 antagonist (BD-1063) and also Gabapentin, as well as determine its possible role as an antagonist of sigma-1 receptors. The anti-hyperalgesic effects of Gabapentin (10.0, 17.8, 31.6, 56.2 and 100mg/kg, s.c.), BD-1063 (5.6, 10.0, 17.8, 31.6 and 56.2mg/kg, s.c.) and NMIN (31.6, 10.0, 316mg/kg and 562mg/kg, s.c.) were determined after single-doses, using the von Frey test in the CCI model. NMIN had the same efficacy as BD-1063, but both show less efficacy than Gabapentin. In an analysis of pharmacological potency, the ED50 were compared with it being found that BD-1063 is the most potent drug, followed by Gabapentin and NMIN. The anti-hyperalgesic effect of NMIN on CCI rats was reversed by (+)-pentazocine (s.c. route) and by PRE-084 (i.t. route), both sigma-1 agonists. Furthermore, NMIN reversed the hyperalgesic effect of PRE-084 in naïve rats. These results suggest that NMIN has an anti-hyperalgesic effect on the CCI model, and that one of its mechanisms of action is as a sigma-1 antagonist, being a significant role the blocking of these receptors at the spinal level.


Automated locomotor activity monitoring as a quality control assay for mass-reared tephritid flies.
The Sterile Insect Technique (SIT) requires vast numbers of consistently high quality insects to be produced over long periods. Quality control (QC) procedures are critical to effective SIT, both providing quality assurance and warning of operational deficiencies. We here present a potential new QC assay for mass rearing of Queensland fruit flies (Bactrocera tryoni Froggatt) for SIT; locomotor activity monitoring. We investigated whether automated locomotor activity monitors (LAMs) that simply detect how often a fly passes an infrared sensor in a glass tube might provide similar insights but with much greater economy. Activity levels were generally lower for females than for males, and declined over five days in the monitor for both sexes. Female activity levels were not affected by irradiation, but males irradiated at 60 or 70 Gy had reduced activity levels compared with unirradiated controls. We also found some evidence that mild heat shock of pupae results in adults with reduced activity. LAM offers a convenient, effective and economical assay to probe such changes.


Trends of Gallbladder Cancer in Jordan Over 2 Decades: Where Are We?
The prevalence of gallbladder cancer (GBC) varies between different parts of the world. This study is a review of literature and an update of a previously published study conducted in our university and aims to reassess the incidence of GBC over the past 2 decades. We conducted a retrospective study between 2002 and 2016. Data regarding demographics, clinical presentation, risk factors, histopathology, investigations, and treatments were obtained. A diagnosis of GBC established during surgery or primarily detected in the surgical specimen was classified as incidental. Of 11 391 cholecystectomies performed, 31 cases (0.27%) of GBC were found. The mean age of patients with GBC was 68 years (43-103 years), 74% were women. The annual incidence of GBC was 0.2/100 000 (men: 0.1/100 000; women: 0.3/100 000). Biliary colic and acute cholecystitis were the main presentations. Diagnosis of GBC was "incidental" in 67% of cases. About 75% of patients with GBC had gallstones, 13% had polyps, and 3% had porcelain gallbladder. Adenocarcinoma was the dominant (87%) histologic type. The GBC rate in our region, similar to others parts of the world, is still low and has not changed over the past 2 decades. This study consolidates the previously published recommendations regarding the high index of suspicion of GBC in elderly with cholelithiasis.


[Rubino's reaction].
Some aspects related to the Rubino's reaction are commented, viewing both a better identification of the involved elements and the arousing of the investigators interest for this test. The following aspects are discussed: 1) the influence of temperature upon the elements of the reaction; 2) Correlation between Rubino's reaction and: a) humoral alterations observed in leprosy; b) cell mediated immunity; 3) variations of the serologic reactivity; 4) prognosis value and healing criterion.


Prenatal diagnosis of multiple acyl-CoA dehydrogenase deficiency: association with elevated alpha-fetoprotein and cystic renal changes.
We report the occurrence of multiple acyl-CoA dehydrogenase deficiency (MADD) in two consecutive pregnancies in a young, Caucasian, non-consanguineous couple. In the first pregnancy, the maternal serum alpha-fetoprotein was elevated. A sonogram showed growth delay, cystic renal disease, and oligohydramnios; the parents decided to terminate the pregnancy. Postmortem examination confirmed the cystic renal disease and showed hepatic steatosis, raising the suspicion of a metabolic disorder. The diagnosis of MADD was made by immunoblot studies on cultured fibroblasts. In the subsequent pregnancy, a sonogram at 15 weeks' gestation showed an early growth delay but normal kidneys. The maternal serum and amniotic fluid concentrations of alpha-fetoprotein were elevated, and the amniotic fluid acylcarnitine profile was consistent with MADD. In vitro metabolic studies on cultured amniocytes confirmed the diagnosis. A follow-up sonogram showed cystic renal changes. These cases provide additional information regarding the evolution of renal changes in affected fetuses and show a relationship with elevated alpha-fetoprotein, which may be useful in counseling the couple at risk. MADD should be considered in the differential diagnosis of elevated alpha-fetoprotein and cystic renal disease. Early growth delay may be an additional feature.


Extracorporeal Photopheresis for Bronchiolitis Obliterans Syndrome After Lung Transplantation.
Lung transplantation is a therapeutic option for select patients with end-stage lung disease. However, successful lung transplantation is hampered by chronic lung allograft dysfunction, in particular bronchiolitis obliterans syndrome (BOS). Although there is no approved or standard treatment for BOS, which may have several distinct phenotypes, extracorporeal photopheresis (ECP) has shown promising results in patients who develop BOS refractory to azithromycin treatment. We reviewed all relevant clinical data indexed on PubMed from 1987 to 2017 to evaluate the role of ECP in patients with BOS. Seven small studies investigated the immunomodulatory effects of ECP in patients after solid organ transplant, and 12 studies reported clinical data specific to ECP therapy for BOS. Studies indicate that ECP triggers an apoptotic cellular cascade that exerts various immunomodulatory effects mediated via increases in anti-inflammatory cytokines, a decrease in proinflammatory cytokines, and an increase in tolerogenic regulatory T cells. Clinical evidence derived from relatively small single-center studies suggests that ECP therapy is associated with improvement or stabilization in lung function and sustainable, statistically significant, decreases in the rate of lung function decline in patients with BOS. Additionally, when adverse event data were reported, ECP was generally well tolerated. None of the comparative studies were randomized. Immunomodulation mediated via ECP is a rational therapeutic option that may improve clinical outcomes in patients with BOS, particularly in the context of in-depth patient phenotyping as part of a stratified approach to treatment; good quality randomized controlled trials are needed to confirm observational findings.


Fractionation of crude extracts from controlled dried and commercially available stem bark of Juglans regia and their antimicrobial effects.
The current research investigates the anti-microbial activities of methanol, ethyl acetate, n-hexane , n-butanol and water extracted samples from controlled dried and commercial bark of walnut (Juglans regia) against five bacterial (Staphylococcus aureus, Bacilus subtilis, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonia and one fungal pathogenic specie (Candida albicans) by discs diffusion susceptibility assay using 0.5 and 1mg disc-1 concentrations. Our results revealed that all the extracts from controlled dried and commercial bark of walnut showed varying degrees of antimicrobial activities. Ethyl acetate fraction from both sources exhibited maximum activity against all tested microbial species followed by n-butanol and crude methanolic extraxt. N-hexane and aqueous extracted samples from controlled dried bark reduced the growth of all studied microbes except Staphylococcus aureus in case of commercial available bark. Aqueous extracted sample showed inhibitory effects against all tested microbes except Candida albicans respectively in case of commercial bark. The most susceptible gram positive bacteria were S. aureus while Bacillus subtilis was the most resistant one. Among Gram negative bacteria, Pseudomonas aeruginosa was the most susceptibility while Klebsella pneumonia showed some resistively. Compared to commercial bark samples, controlled dried bark extract and fractions were found to be more active in reducing the growth of all the tested microbes at both concentrations.


Rational engineering of an erythropoietin fusion protein to treat hypoxia.
Erythropoietin enhances oxygen delivery and reduces hypoxia-induced cell death, but its pro-thrombotic activity is problematic for use of erythropoietin in treating hypoxia. We constructed a fusion protein that stimulates red blood cell production and neuroprotection without triggering platelet production, a marker for thrombosis. The protein consists of an anti-glycophorin A nanobody and an erythropoietin mutant (L108A). The mutation reduces activation of erythropoietin receptor homodimers that induce erythropoiesis and thrombosis, but maintains the tissue-protective signaling. The binding of the nanobody element to glycophorin A rescues homodimeric erythropoietin receptor activation on red blood cell precursors. In a cell proliferation assay, the fusion protein is active at 10-14 M, allowing an estimate of the number of receptor-ligand complexes needed for signaling. This fusion protein stimulates erythroid cell proliferation in vitro and in mice, and shows neuroprotective activity in vitro. Our erythropoietin fusion protein presents a novel molecule for treating hypoxia.


Role of patient preferences in clinical practice guidelines: a multiple methods study using guidelines from oncology as a case.
Many treatment decisions are preference-sensitive and call for shared decision-making, notably when benefits are limited or uncertain, and harms impact quality of life. We explored if clinical practice guidelines (CPGs) acknowledge preference-sensitive decisions in how they motivate and phrase their recommendations. We performed a qualitative analysis of the content of CPGs and verified the results in semistructured interviews with CPG panel members. Dutch oncology CPGs issued in 2010 or later, concerning primary treatment with curative intent. 14 CPG panel members. For treatment recommendations from six CPG modules, two researchers extracted the following: strength of recommendation in terms of the Grading of Recommendations Assessment, Development and Evaluation and its consistency with the CPG text; completeness of presentation of benefits and harms; incorporation of patient preferences; statements on the panel's benefits-harm trade-off underlying recommendation; and advice on patient involvement in decision-making. We identified 32 recommendations, 18 were acknowledged preference-sensitive decisions. Three of 14 strong recommendations should have been weak based on the module text. The reporting of benefits and harms, and their probabilities, was sufficiently complete and clear to inform the strength of the recommendation in one of the six modules only. Numerical probabilities were seldom presented. None of the modules presented information on patient preferences. CPG panel's preferences were not made explicit, but appeared to have impacted 15 of 32 recommendations. Advice to involve patients and their preferences in decision-making was given for 20 recommendations (14 weak). Interviewees confirmed these findings. Explanations for lack of information were, for example, that clinicians know the information and that CPGs must be short. Explanations for trade-offs made were cultural-historical preferences, compliance with daily care, presumed role of CPGs and lack of time. The motivation and phrasing of CPG recommendations do not stimulate choice awareness and a neutral presentation of options, thus hindering shared decision-making.


Elucidation of GlcNAc-binding properties of type III intermediate filament proteins, using GlcNAc-bearing polymers.
Vimentin, desmin, glial fibrillary acidic protein (GFAP) and peripherin belong to type III intermediate filament family and are expressed in mesenchymal cells, skeletal muscle cells, astrocytes and peripheral neurons, respectively. Vimentin and desmin possess N-acetyl-d-glucosamine (GlcNAc)-binding properties on cell surfaces. The rod II domain of these proteins is a GlcNAc-binding site, which also exists in GFAP and peripherin. However, the GlcNAc-binding activities and behaviors of these proteins remain unclear. Here, we characterized the interaction and binding behaviors of these proteins, using various well-defined GlcNAc-bearing polymers synthesized by radical polymerization with a reversible addition-fragmentation chain transfer reagent. The small GlcNAc-bearing polymers strongly interacted with HeLa cells through vimentin expressed on the cell surface and interacted with vimentin-, desmin-, GFAP- and peripherin-transfected vimentin-deficient HeLa cells. These proteins present high affinity to GlcNAc-bearing polymers, as shown by surface plasmon resonance. These results show that type III intermediate filament proteins possess GlcNAc-binding activities on cell surfaces. These findings provide important insights into novel cellular functions and physiological significance of type III intermediate filaments.


Proteomics of genetically engineered mouse mammary tumors identifies fatty acid metabolism members as potential predictive markers for cisplatin resistance.
In contrast to various signatures that predict the prognosis of breast cancer patients, markers that predict chemotherapy response are still elusive. To detect such predictive biomarkers, we investigated early changes in protein expression using two mouse models for distinct breast cancer subtypes who have a differential knock-out status for the breast cancer 1, early onset (Brca1) gene. The proteome of cisplatin-sensitive BRCA1-deficient mammary tumors was compared with that of cisplatin-resistant mammary tumors resembling pleomorphic invasive lobular carcinoma. The analyses were performed 24 h after administration of the maximum tolerable dose of cisplatin. At this time point, drug-sensitive BRCA1-deficient tumors showed DNA damage, but cells were largely viable. By applying paired statistics and quantitative filtering, we identified highly discriminatory markers for the sensitive and resistant model. Proteins up-regulated in the sensitive model are involved in centrosome organization, chromosome condensation, homology-directed DNA repair, and nucleotide metabolism. Major discriminatory markers that were up-regulated in the resistant model were predominantly involved in fatty acid metabolism, such as fatty-acid synthase. Specific inhibition of fatty-acid synthase sensitized resistant cells to cisplatin. Our data suggest that exploring the functional link between the DNA damage response and cancer metabolism shortly after the initial treatment may be a useful strategy to predict the efficacy of cisplatin.


An Exaggerated Immune Response in Female BALB/c Mice Controls Initial Toxoplasma gondii Multiplication but Increases Mortality and Morbidity Relative to Male Mice.
Studies indicate that female mice are more susceptible to T. gondii infection, as defined by higher mortality rates in comparison to male mice. However, whether this is due to an inability to control initial parasite multiplication or due to detrimental effects of the immune system has not been determined. Therefore, the following studies were undertaken to determine the influence of sex on early parasite multiplication and the immune response during T. gondii infection and to correlate this with disease outcome. Early parasite replication was studied through applying an in vivo imaging system (IVIS) with luciferase expressing T. gondii. In parallel immunological events were studied by cytometric bead array to quantify key immunological mediators. The results confirmed the previous findings that female mice are more susceptible to acute infection, as determined by higher mortality rates and weight loss compared with males. However, conflicting with expectations, female mice had lower parasite burdens during the acute infection than male mice. Female mice also exhibited significantly increased production of Monocyte Chemoattractant Protein-1 (MCP-1), Interferon (IFN)-γ, and Tumour Necrosis Factor (TNF)-α than male mice. MCP-1 was found to be induced by T. gondii in a dose dependent manner suggesting that the observed increased levels detected in female mice was due to a host-mediated sex difference rather than due to parasite load. However, MCP-1 was not affected by physiological concentration of estrogen or testosterone, indicating that MCP-1 differences observed between the sexes in vivo are due to an as yet unidentified intermediary factor that in turn influences MCP-1 levels. These results suggest that a stronger immune response in female mice compared with male mice enhances their ability to control parasite replication but increases their morbidity and mortality.


A thermal activated and differential self-calibrated flexible epidermal biomicrofluidic device for wearable accurate blood glucose monitoring.
This paper reports a flexible electronics-based epidermal biomicrofluidics technique for clinical continuous blood glucose monitoring, overcoming the drawback of the present wearables, unreliable measurements. A thermal activation method is proposed to improve the efficiency of transdermal interstitial fluid (ISF) extraction, enabling extraction with a low current density to notably reduce skin irritation. An Na+ sensor and a correction model are proposed to eliminate the effect of individual differences, which leads to fluctuations in the amount of ISF extraction. An electrochemical sensor with a 3D nanostructured working electrode surface is designed to enable precise in situ glucose measurement. A differential structure is proposed to eliminate the effect of passive perspiration, which leads to inaccurate blood glucose prediction. Fabrications of the epidermal biomicrofluidic device including formation of flexible electrodes, nanomaterial modification, and enzyme immobilization are fully realized by inkjet printing to enable facile manufacturing with low cost, which benefits practical production.


Advising patients about breast implant associated anaplastic large cell lymphoma.
The United States Food and Drug Administration (FDA) recently recognized the necessity for improved patient education and public awareness of breast implant-associated anaplastic large cell lymphoma (BIA-ALCL). BIA-ALCL is an uncommon T-cell lymphoma that can arise years after placement of a textured breast implant or tissue expander. Patients and providers would greatly benefit from a broader awareness of BIA-ALCL and how it presents with both common and rare symptoms. Disease awareness can be improved with specific steps moving forward. This includes improved counseling about annual recommendations from the FDA, standard of care guidelines, current estimates of disease prevalence, and manufacturer-specific risk for developing the malignancy. Informed consent for implant-based breast surgery should include a discussion of BIA-ALCL, symptoms, inherent risks and alternatives. Consideration should be given for reasonable efforts to retroactively contact patients with textured implants to inform them of recent regulatory actions as well as the risk of developing the disease and the signs and symptoms to watch for. Where risk can be reduced, all steps should be taken to ensure patient safety. In keeping with our commitment to patient safety, the strategies moving forward for implant-based breast surgery should be marked by patient awareness, physician vigilance, and defined by evidence-based diagnosis and treatment.


Using the humanities to strengthen the concept of professionalism among first-professional year pharmacy students.
To engage pharmacy students at the McWhorter School of Pharmacy in an authentic discussion of professionalism early in their education. A booklet was prepared that included several classic short stories and essays that dealt with professionalism. This booklet was sent to all entering students in the class of 2008 and 2009 during the summer prior to their first-professional year of the PharmD program. The stories and essays were discussed in small groups with faculty facilitation during orientation when the students first arrived on campus. A survey instrument was created and administered to assess the impact of this innovative approach to enhancing professionalism. The program was well received and engaged our pharmacy students in a productive discussion on professionalism. Both classes' mean scores on survey items related that the students were engaged in the discussion of professionalism. Survey results pertaining to professional behavior also indicated increased awareness of the importance of professionalism. Enhancing professionalism requires a culture change that necessitates addressing professionalism at its core, a calling to serve, in a persistent and continual manner. Requiring students to read and think about professionalism in a novel way, before they even begin their first-professional year of pharmacy school, appears to be an effective approach to nurturing/encouraging professionalism.


Epstein-Barr virus negative clonal plasma cell proliferations and lymphomas in peripheral T-cell lymphomas: a phenomenon with distinctive clinicopathologic features.
Clonal B-cell populations have been described in peripheral T-cell lymphomas (PTCL) as secondary Epstein-Barr virus (EBV) driven B-cell expansions that may evolve to an overt B-cell lymphoma. EBV-negative B-cell proliferations associated with T-cell lymphomas are uncommon and not well characterized. We studied 15 patients who developed an EBV-negative B-cell proliferation or malignant lymphoma associated with PTCL. The T-cell tumors were 8 PTCL, not otherwise specified, 4 angioimmunoblastic T-cell lymphomas, and 3 cutaneous PTCL. The B-cell component was intermingled with the PTCL in all patients and it was classified as clonal/monotypic plasma cell proliferation in 8 lesions, clonal/monotypic large B-cell proliferation in 4 patients, and B-cell lymphoma with plasmacytic/plasmablastic differentiation in 3 patients. Two patients had 2 clonally unrelated plasma cell proliferations associated with the same PTCL. All cases showed cytoplasmic Ig light chain restriction. Clonal IgH and T-cell receptor rearrangements were detected in 11/12 and 11/13 cases examined, respectively. EBV, cytomegalovirus, and HHV-8 were not observed in any of the examined cases. Sequential samples in 7 patients showed persistence of the PTCL and the B-cell component in 4, the PTCL without the B-cell lymphoma in 2, and progression of the B-cell neoplasm in 1. Patients followed an aggressive clinical course similar to conventional PTCL. In conclusion, EBV-negative clonal or mononotypic B-cell proliferations in patients with PTCL present with a spectrum of lesions ranging from plasma cell proliferations to overt lymphomas with plasmacytic/plasmablastic features. The distinctive features of these patients suggest that these lesions represent a specific phenomenon in PTCL.


A new approach to quantify the adaptive potential of gene expression variation in gymnosperms.
Variation in patterns of gene expression contributes to phenotypic diversity and can ultimately predict adaptive responses. However, in many cases, the consequences of regulatory mutations on patterns of gene expression and ultimately phenotypic differences remain elusive. A standard way to study the genetic architecture of expression variation in model systems has been to map gene expression variation to genetic loci (Fig. 1a). At the same time, in many nonmodel species, especially for long-lived organisms, controlled crosses are not feasible. If we are to expand our understanding of the role of regulatory mutations on phenotypes, we need to develop new methodologies to study species under ecologically relevant conditions. In this issue of Molecular Ecology, Verta et al. (2013) present a new approach to analyse gene expression variation and regulatory networks in gymnosperms (Fig. 1b). They capitalized on the fact that gymnosperm seeds contain an energy storage tissue (the megagametophyte) that is directly derived from a single haploid cell (the megaspore). The authors identified over 800 genes for which expression segregated in this maternally inherited haploid tissue. Based on the observed segregation patterns, these genes (Mendelian Expression Traits) are most probably controlled by biallelic variants at a single locus. Most of these genes also belonged to different regulatory networks, except for one large group of 180 genes under the control of a putative trans-acting factor. In addition, the approach developed here may also help to uncover the effect of rare recessive mutations, which usually remain hidden in a heterozygous state in diploid individuals. The appeal of the work by Verta et al. (2013) to study gene expression variation is in its simplicity, which circumvents several of the hurdles behind traditional expression quantitative trait locus (eQTL) studies, and could potentially be applied to a large number of species.


Geraniol protects against cyclophosphamide-induced hepatotoxicity in rats: Possible role of MAPK and PPAR-γ signaling pathways.
Cyclophosphamide (CP) is one of the famous anti-cancer drugs. However, CP-induced hepatotoxicity is a dose-limiting side effect. The present study aimed to investigate the potential protective effect of geraniol (GOH), the main ingredient of Palmarosa oil and rose oil, against CP-induced hepatotoxicity in rats. Results showed that CP provoked a marked elevation in serum levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase. In addition, oxidative stress was significantly boosted in CP-treated rats as compared to control rats. On the other hand, GOH (200 mg/kg, p.o.) administration attenuated CP-evoked disturbances in the above-mentioned parameters. Moreover, histopathological aberrations in CP-treated rats were significantly ameliorated in GOH-treated rats. GOH markedly abrogated CP-induced inflammation via decreasing the protein expression of nuclear factor-kappa B, inducible nitric oxide synthase and cyclo-oxygenase 2, as well as reducing the levels of pro-inflammatory cytokines in CP-treated rats. CP induced activation of MAPK; p38 and JNK and diminished PPAR-γ protein expression. GOH effectively reversed all these effects. In conclusion, GOH is suggested to be a potential candidate for attenuation of CP-induced hepatotoxicity. This effect is attributed to its antioxidant and anti-inflammatory activities, as well as, modulation of MAPK and PPAR-γ signaling pathways.


Diagnosing and Managing Breast Disease During Pregnancy and Lactation.
Although carcinoma of the breast complicates 1:3000 deliveries in the US, most breast conditions unique to pregnancy and lactation are benign--for example, lactating adenoma, galactocele, gigantomastia, and benign bloody nipple discharge. Nevertheless, malignancy must be excluded by a thorough work-up, including breast biopsy if indicated; "watchful waiting" when a breast mass is discovered is no more appropriate than in a nonpregnant patient. During lactation, the major problems encountered often are part of a spectrum of inflammatory and infectious complications. Nasopharyngeal organisms from the infant are usually the source of breast infections in lactating women. Keeping the breast empty of milk promotes healing by helping to drain the culture medium that is facilitating growth of organisms. Hence, the earlier recommendations that breast-feeding cease during mastitis have been superseded by the knowledge that breast-feeding is generally not harmful to the infant and may speed resolution of the infectious process. The diagnosis and management of pregnancy-associated breast cancer (PABC) is reviewed. Pregnancy-associated masses are usually discovered by patient self-examination, and the clinician should proceed to fine-needle aspiration or biopsy, rather than mammography, which has poor sensitivity during pregnancy and lactation because of increased breast density. Management of a new breast mass in pregnancy should maximize diagnostic accuracy and minimize the chances of missing PABC, yet avoid harm to the fetus or interruption of lactation.


The Clinical Effectiveness and Cost-Effectiveness of Lamotrigine in Borderline Personality Disorder: A Randomized Placebo-Controlled Trial.
The authors examined whether lamotrigine is a clinically effective and cost-effective treatment for people with borderline personality disorder. This was a multicenter, double-blind, placebo-controlled randomized trial. Between July 2013 and November 2016, the authors recruited 276 people age 18 or over who met diagnostic criteria for borderline personality disorder. Individuals with coexisting bipolar affective disorder or psychosis, those already taking a mood stabilizer, and women at risk of pregnancy were excluded. A web-based randomization service was used to allocate participants randomly in a 1:1 ratio to receive either an inert placebo or up to 400 mg/day of lamotrigine. The primary outcome measure was score on the Zanarini Rating Scale for Borderline Personality Disorder (ZAN-BPD) at 52 weeks. Secondary outcome measures included depressive symptoms, deliberate self-harm, social functioning, health-related quality of life, resource use and costs, side effects of treatment, and adverse events. A total of 195 (70.6%) participants were followed up at 52 weeks, at which point 49 (36%) of those in the lamotrigine group and 58 (42%) of those in the placebo group were taking study medication. The mean ZAN-BPD score was 11.3 (SD=6.6) among those in the lamotrigine group and 11.5 (SD=7.7) among those in the placebo group (adjusted difference in means=0.1, 95% CI=-1.8, 2.0). There was no evidence of any differences in secondary outcomes. Costs of direct care were similar in the two groups. The results suggest that treating people with borderline personality disorder with lamotrigine is not a clinically effective or cost-effective use of resources.


Hyperbaric oxygen therapy: Can it be a novel supportive therapy in COVID-19?
The coronavirus disease 2019 (COVID-19) is a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV2). Although 85% of infected patients remain asymptomatic, 5% show severe symptoms such as hypoxaemic respiratory failure and multiple end organ dysfunction (MODS) requiring intensive care unit (ICU) admission with a mortality rate of about 2.8%. Since a definitive treatment is yet to be identified, preventive and supportive strategies remain the mainstay of management. Supportive measures such as oxygen therapy with nasal cannula, face mask, noninvasive ventilation, mechanical ventilation and even extreme measures such as extracorporeal membrane oxygenation (ECMO) fail to improve oxygenation in some patients. Hence, hyperbaric oxygen therapy (HBOT) has been proposed as a supportive strategy to improve oxygenation in COVID-19 patients. HBOT is known to increase tissue oxygenation by increasing the amount of dissolved oxygen in plasma. HBOT also mitigates tissue inflammation thus reducing the ill effects of cytokine storm in COVID-19 patients. Though there is limited literature available on HBOT in COVID-19 patients, considering the present need for additional supportive therapy to improve oxygenation, HBOT has been proposed as a novel supportive treatment in COVID-19 patients.


miRNA suppression of a Notch repressor directs non-neuronal fate in Drosophila mechanosensory organs.
Although there is abundant evidence that individual microRNA (miRNA) loci repress large cohorts of targets, large-scale knockout studies suggest that most miRNAs are phenotypically dispensable. Here, we identify a rare case of developmental cell specification that is highly dependent on miRNA control of an individual target. We observe that binary cell fate choice in the Drosophila melanogaster peripheral sensory organ lineage is controlled by the non-neuronally expressed mir-279/996 cluster, with a majority of notum sensory organs exhibiting transformation of sheath cells into ectopic neurons. The mir-279/996 defect phenocopies Notch loss of function during the sheath-neuron cell fate decision, suggesting the miRNAs facilitate Notch signaling. Consistent with this, mir-279/996 knockouts are strongly enhanced by Notch heterozygosity, and activated nuclear Notch is impaired in the miRNA mutant. Although Hairless (H) is the canonical nuclear Notch pathway inhibitor, and H heterozygotes exhibit bristle cell fate phenotypes reflecting gain-of-Notch signaling, H/+ does not rescue mir-279/996 mutants. Instead, we identify Insensible (Insb), another neural nuclear Notch pathway inhibitor, as a critical direct miR-279/996 target. Insb is posttranscriptionally restricted to neurons by these miRNAs, and its heterozygosity strongly suppresses ectopic peripheral nervous system neurons in mir-279/996 mutants. Thus, proper assembly of multicellular mechanosensory organs requires a double-negative circuit involving miRNA-mediated suppression of a Notch repressor to assign non-neuronal cell fate.


Water dynamics in silica nanopores: the self-intermediate scattering functions.
The dynamics of water molecules confined in approximately cylindrical silica nanopores is investigated using molecular simulation. The model systems are pores of diameter varying between 20 and 40 Å containing water at room temperature and at full hydration, prepared using grand canonical Monte Carlo simulation. Water dynamics in these systems is studied via molecular dynamics simulation. The results of the basic characterization of these systems have been reported in A. A. Milischuk and B. M. Ladanyi [J. Chem. Phys. 135, 174709 (2011)]. The main focus of the present study is the self-intermediate scattering function (ISF), F(S)(Q, t), of water hydrogens, the observable in quasi-elastic neutron scattering experiments. We investigate how F(S)(Q, t) depends on the pore diameter, the direction and magnitude of the momentum transfer Q, and the proximity of water molecules to the silica surface. We also study the contributions to F(S)(Q, t) from rotational and translational motions of water molecules and the extent of rotation-translation coupling present in F(S)(Q, t). We find that F(S)(Q, t) depends strongly on the pore diameter and that this dependence is due mainly to the contributions to the ISF from water translational motion and can be attributed to the decreased mobility of water molecules near the silica surface. The relaxation rate depends on the direction of Q and is faster for Q in the axial than in the radial direction. As the magnitude of Q increases, this difference diminishes but does not disappear. We find that its source is mainly the anisotropy in translational diffusion at low Q and in molecular reorientation at higher Q values.


Engineering lanthipeptides by introducing a large variety of RiPP modifications to obtain new-to-nature bioactive peptides.
Natural bioactive peptide discovery is a challenging and time-consuming process. However, advances in synthetic biology are providing promising new avenues in peptide engineering that allow for the design and production of a large variety of new-to-nature peptides with enhanced or new bioactivities, using known peptides as templates. Lanthipeptides are ribosomally synthesized and post-translationally modified peptides (RiPPs). The modularity of post-translational modification (PTM) enzymes and ribosomal biosynthesis inherent to lanthipeptides enables their engineering and screening in a high-throughput manner. The field of RiPPs research is rapidly evolving, with many novel PTMs and their associated modification enzymes being identified and characterized. The modularity presented by these diverse and promiscuous modification enzymes has made them promising tools for further in vivo engineering of lanthipeptides, allowing for the diversification of their structures and activities. In this review, we explore the diverse modifications occurring in RiPPs and discuss the potential applications and feasibility of combining various modification enzymes for lanthipeptide engineering. We highlight the prospect of lanthipeptide- and RiPP-engineering to produce and screen novel peptides, including mimics of potent non-ribosomally produced antimicrobial peptides (NRPs) such as daptomycin, vancomycin, and teixobactin, which offer high therapeutic potential.


Double cation adduction in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of electron deficient anthraquinone derivatives.
Six anthraquinone derivatives were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS). Clear (pseudo) molecular ions were observed for all the compounds. Interestingly, for some derivatives, strong ions with double cation adduction were also recorded in the positive mode. It is remarkable that all these ions are singly charged. In this work, possible mechanisms for the double cation adduction were investigated and discussed. It appears that the double cation adduction was due to the electron deficient nature of the derivatives, and formed by taking up two singly charged cations and one electron. Substituents on the anthraquinone ring were found to have a significant effect on the double cation adduction. In contrast, no considerable influence of the acidity of MALDI matrix/solution was observed, even on the double proton adduction. Furthermore, it was demonstrated that double cation adduction might occur in the MALDI gas-phase plume. In addition to the anthraquinones, three more electron deficient compounds of different types, i.e. a perylene bisimide derivative (PB), 3,7-decanoylamino-4,8-dihydrobenzo[1,2-b:4,5-b']dithiophene-4,8-dione (TQ) and 6,6-phenyl C61-butyric acid methyl ester (PCBM), were also analyzed with MALDI TOF MS. The results indicate that the 'abnormal' double cation adduction might be a 'normal' phenomenon in the MALDI TOF MS analysis of many electron deficient compounds.


Impact of vitamin D treatment on 25 hydroxy vitamin D levels and insulin homeostasis in obese African American adolescents in a randomized trial.
Obesity is affecting children in epidemic proportions in the United States with nearly 25% of children being obese. Consequences of obesity including dyslipidemia, type 2 diabetes and cardiovascular disease are leading to morbidity at younger ages. Parallel to the obesity and diabetes epidemics, the prevalence of vitamin D deficiency has reached very high levels and has been associated with insulin resistance and dyslipidemia. Studies exploring the impact of vitamin D repletion on insulin sensitivity and dyslipidemia in children are sparse.The aim of this study was to determine the impact of treatment with vitamin D (ergocalciferol) in obese African American (AA) children on vitamin D levels and insulin secretion and sensitivity. This pilot study was conducted in a tertiary care Pediatric Emergency Department (ED). African American obese children (n = 29; 22 female) 13-17 y, with 25-hydroxy vitamin D level [25(OH)D] <20 ng/ml, were randomized to receive either 50,000 IU vitamin D2/week or a placebo for 12 weeks. Pre- and post- oral glucose tolerance testing with glucose and insulin levels drawn at 0, 30, 60, 90 and 120 min were performed. Pre/post intervention lipid profiles and calcium levels were also evaluated. There was no difference in serum 25(OH)D between groups at baseline. Follow-up 25(OH)D level was greater in the treatment vs. placebo group, and significantly increased from baseline in the treatment group only. However, there was no difference between groups in baseline vs. follow-up insulin- or lipid-related parameters. Follow-up serum 25(OH)D was positively correlated with fasting insulin and high-density lipoprotein (HDL) level in the vitamin D treated group only. While serum 25(OH)D levels in obese AA teens increased adequately with vitamin treatment for 12 weeks and correlated with fasting insulin, it did not significantly impact insulin secretion or sensitivity. Larger studies are required over a longer period of time to confirm and explore the reasons for this finding.


Heart failure with preserved ejection fraction.
Heart failure with preserved ejection fraction (HFpEF) has recently emerged as a major cause of cardiovascular morbidity and mortality. Contrary to initial beliefs, HFpEF is now known to be as common as heart failure with reduced ejection fraction (HFrEF) and carries an unacceptably high mortality rate. With a prevalence that has been steadily rising over the past two decades, it is very likely that HFpEF will represent the dominant heart failure phenotype over the coming few years. The scarcity of trials in this semi-discrete form of heart failure and lack of unified enrolment criteria in the studies conducted to date might have contributed to the current absence of specific therapies. Understanding the epidemiological, pathophysiological and molecular differences (and similarities) between these two forms of heart failure is cornerstone to the development of targeted therapies. Carefully designed studies that adhere to unified diagnostic criteria with the recruitment of appropriate controls and adoption of practical end-points are urgently needed to help identify effective treatment strategies.


Multistate Dynamic Pathways for Anisotropic Colloidal Assembly and Reconfiguration.
We report the controlled interfacial assembly and reconfiguration of rectangular prism colloidal particles between microstructures of varying positional and orientational order including stable, metastable, and transient states. Structurally diverse states are realized by programming time dependent electric fields that mediate dipolar interactions determining particle position, orientation, compression, and chaining. We identify an order parameter set that defines each state as a combination of the positional and orientational order. These metrics are employed as reaction coordinates to capture the microstructure evolution between initial and final states upon field changes. Assembly trajectory manifolds between states in the low-dimensional reaction coordinate space reveal a dynamic pathway map including information about pathway accessibility, reversibility, and kinetics. By navigating the dynamic pathway map, we demonstrate reconfiguration between states on minute time scales, which is practically useful for particle-based materials processing and device responses. Our findings demonstrate a conceptually general approach to discover dynamic pathways as a basis to control assembly and reconfiguration of self-organizing building blocks that respond to global external stimuli.


Rac1 signalling in the Drosophila larval cellular immune response.
The Drosophila larval cellular immune response involves cells (hemocytes) that can be recruited from a hematopoietic organ located behind the brain, as well as a sessile population of cells found just underneath the larval cuticle arranged in a segmental pattern. By using two Rac1 GTPase effector-loop mutants together with epistasis studies, we show that Rac1 requires the Drosophila melanogaster Jun N-terminal kinase Basket (Bsk), as well as stable actin formation to recruit the sessile hemocyte population. We show that actin stabilization is necessary for Rac1-induced hemocyte activation by lowering cofilin (encoded by the twinstar gene tsr) expression in blood cells. Removing Bsk by RNAi suppressed Rac1-induced release of sessile hemocytes. RNAi against Bsk also suppressed Rac1 induction of lamellocytes, a specialized population of hemocytes necessary for the encapsulation of invading pathogens. Furthermore, Rac1 and Bsk are involved in regulating the formation of actin- and focal adhesion kinase (FAK)-rich placodes in hemocytes. Lastly, Rac1 and Bsk are both required for the proper encapsulation of eggs from the parasitoid wasp Leptipolina boulardi. From these data we conclude that Rac1 induces Bsk activity and stable actin formation for cellular immune activation, leading to sessile hemocyte release and an increase in the number of circulating hemocytes.


Interaction of carbocyanine dyes with DNA: synthesis and spectroscopic studies.
Six carbocyanine dyes have been synthesized and their interactions with DNA have been investigated for their prospective use as fluorescent markers in molecular biology. The noncovalent binding of the compounds with DNA was explored by fluorescence spectroscopy, ultraviolet/visible spectrophotometry, and photobleaching. The electron-withdrawing ability of the substituents in N-position of free thio- and selenocarbocyanines strongly affected their photostabilities. Changes in the experimental conditions, such as the presence or absence of oxygen, had an impact on the rate of photobleaching. In the presence of DNA, absorbance photofading and fluorescence photobleaching occurred. It appears that the reduction of fluorescence intensity was due to quenching of the dye fluorescence by interaction with DNA.


Degenerate four-wave mixing in plasmas.
We report an analysis of degenerate four-wave mixing in uniform plasmas and show that the magnitude of the third-order susceptibility (varying as lambda(2)) can be comparable to (10(-11) esu for lambda = 10.6 microm) or orders of magnitude larger (10(-3) esu for lambda= 10 cm) than the corresponding susceptibility in typical nonlinear materials. The nonlinear contribution to the macroscopic current resulting from the ponderomotive force is generated from the two-component fluid model by means of a linearized perturbation scheme. Numerical estimates of the nonlinear susceptibility show that relatively large signals can be produced easily without significant problems from competing physical phenomena such as self-focusing or stimulated Brillouin and Raman scattering.


Uncertainty-Aware Blind Image Quality Assessment in the Laboratory and Wild.
Performance of blind image quality assessment (BIQA) models has been significantly boosted by end-to-end optimization of feature engineering and quality regression. Nevertheless, due to the distributional shift between images simulated in the laboratory and captured in the wild, models trained on databases with synthetic distortions remain particularly weak at handling realistic distortions (and vice versa). To confront the cross-distortion-scenario challenge, we develop a unified BIQA model and an approach of training it for both synthetic and realistic distortions. We first sample pairs of images from individual IQA databases, and compute a probability that the first image of each pair is of higher quality. We then employ the fidelity loss to optimize a deep neural network for BIQA over a large number of such image pairs. We also explicitly enforce a hinge constraint to regularize uncertainty estimation during optimization. Extensive experiments on six IQA databases show the promise of the learned method in blindly assessing image quality in the laboratory and wild. In addition, we demonstrate the universality of the proposed training strategy by using it to improve existing BIQA models.


An in vivo and in silico evaluation of the hepatoprotective potential of Gynura procumbens: A promising agent for combating hepatotoxicity.
The liver, the most important metabolic organ of the body, performs a wide variety of vital functions. Hepatic cell injury occurs by the activation of reactive oxygen species (ROS) that are generated by carbon tetrachloride (CCl4), xenobiotics, and other toxic substances through cytochrome P450-dependent steps resulting from the covalent bond formation with lipoproteins and nucleic acids. Observing the urgent state of hepatotoxic patients worldwide, different medicinal plants and their properties can be explored to combat such free radical damage to the liver. In vivo and in silico studies were designed and conducted to evaluate the antioxidant and hepatoprotective properties of Gynura procumbens in rats. Gynura procumbens leaves were collected and extracted using 70% ethanol. The required chemicals CCl4, standard drug (silymarin), and blood serum analysis kits were stocked. The in vivo tests were performed in 140 healthy Wister albino rats of either sex under well-controlled parameters divided into 14 groups, strictly maintaining Institutional Animal Ethics Committee (IEAC) protocols. For the histopathology study, 10% buffered neutral formalin was used for organ preservation. Later the specimens were studied under a fluorescence microscope. In silico molecular docking and absorption, distribution, metabolism, excretion, and toxicity (ADMET) studies were performed, and the results were analyzed statistically. Gynura procumbens partially negate the deleterious effect of carbon tetrachloride on normal weight gain in rats. The elevated level of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), creatinine, LDH, total cholesterol (TC), low-density lipoprotein (LDL), triglycerides (TG), malondialdehyde (MDA), deoxyribonucleic acid (DNA) fragmentation ranges, gamma-glutamyl transferase (γ-GT) in CCl4 treated groups were decreased by both standard drug silymarin and G. procumbens leaf extract. We have found significant & highly significant changes statistically for different doses, here p<0.05 & p<0.01, respectively. On the other hand, G. procumbens and silymarin displayed Statistically significant (p<0.05) and high significant(p<0.01) increased levels of HDL, CAT SOD (here p<0.05 & p<0.01 for different doses) when the treatment groups were compared with the disease control group. Because the therapeutic activity imparted by plants and drugs accelerates the movement of the disturbed pathophysiological state toward the healthy state. In the molecular docking analysis, G. procumbens phytoconstituents performed poorly against transforming growth factor-beta 1 (TGF-β1) compared to the control drug silymarin. In contrast, 26 phytoconstituents scored better than the control bezafibrate against peroxisome proliferator-activated receptor alpha (PPAR-α). The top scoring compounds for both macromolecules were observed to form stable complexes in the molecular dynamics simulations. Flavonoids and phenolic compounds performed better than other constituents in providing hepatoprotective activity. It can, thus, be inferred that the extract of G. procumbens showed good hepatoprotective properties in rats.


Concentration of 25-hydroxyvitamin D from neonatal dried blood spots and the relation to gestational age, birth weight and Ponderal Index: the D-tect study.
Studies have suggested that vitamin D status at birth may be associated with a range of neonatal outcomes. The aim of this study was to assess the association between neonatal 25-hydroxyvitamin D3 (25(OH)D3) concentration and gestational age, birth weight, Ponderal Index and size for gestational age. Neonatal capillary blood stored as dried blood spots was used to assess 25(OH)D3 concentrations among 2686 subjects selected from a random population sub-sample of individuals, born in Denmark from 1 May 1981 to 31 December 2002. There was an inverse association between 25(OH)D3 concentration and gestational age at birth of -0·006 (95 % CI -0·009, -0·003, P<0·001) weeks of gestation per 1 nmol/l increase in 25(OH)D3 concentration. An inverted U-shaped association between 25(OH)D3 and birth weight and Ponderal Index (P=0·04) was found, but no association with size for gestational age was shown. This study suggests that neonatal 25(OH)D3 concentration is associated with anthropometric measures at birth known to be correlated with many subsequent health outcomes such as obesity and type 2 diabetes.


[Acute hepatitis C in the presence of chronic HBV infection].
Acute HCV superinfection was studied in 23 patients with chronic hepatitis B virus infection. HBsAg, anti-HCV (C-100, core, NS3, NS5) were detected in patients' sera at first investigation. Predominant replication of HBV DNA was detected in the sera of 68% patients and HCV RNA in only 24% patients. The clinical course of acute hepatitis C in patients with chronic HBV infection in general corresponded to HCV monoinfection except for more pronounced biochemical shifts and shorter intoxication. The role of HBV and HCV in infectious process is discussed.


[Errors in the diagnosis of acute pancreatitis by means of puncture of the abdominal cavity].
On the basis of the experiences with diagnostic punctures of the abdominal cavity in 91 patients with suspected acute pancreatitis and errors in diagnosis of the disease by data of examination of the exudate, the author recommends as necessary a bacterioscopic examination of the peritoneal exudate and determination of the ammonia level in it.


Deep sequencing of sncRNAs reveals hallmarks and regulatory modules of the transcriptome during Parkinson's disease progression.
Noncoding RNAs have diagnostic and prognostic importance in Parkinson's disease (PD). We studied circulating small noncoding RNAs (sncRNAs) in two large-scale longitudinal PD cohorts (Parkinson's Progression Markers Initiative (PPMI) and Luxembourg Parkinson's Study (NCER-PD)) and modeled their impact on the transcriptome. Sequencing of sncRNAs in 5,450 blood samples of 1,614 individuals in PPMI yielded 323 billion reads, most of which mapped to microRNAs but covered also other RNA classes such as piwi-interacting RNAs, ribosomal RNAs and small nucleolar RNAs. Dysregulated microRNAs associated with disease and disease progression occur in two distinct waves in the third and seventh decade of life. Originating predominantly from immune cells, they resemble a systemic inflammation response and mitochondrial dysfunction, two hallmarks of PD. Profiling 1,553 samples from 1,024 individuals in the NCER-PD cohort validated biomarkers and main findings by an independent technology. Finally, network analysis of sncRNA and transcriptome sequencing from PPMI identified regulatory modules emerging in patients with progressing PD.


[Iatrogenic biliary lesions and stenosis].
Pathogenetic, diagnostic and therapeutic aspects of postoperative bile duct injuries are reviewed. Treatment options are discussed in relation to the time of diagnosis. Lesions detected during the same operation must be immediately repaired through an end-to-end biliary anastomosis or a bilioenteric anastomosis. In limited lesions of the bile duct a T-tube placement should be sufficient. Bile duct lesions recognized postoperatively can be managed through a multimodal surgical, endoscopic, and radiologic approach. In the early postoperative period, surgery is indicated when a complete section of the biliary tract or a severe peritonitis is recognized, or when endoscopic and radiologic treatment has failed. Surgery is also the treatment of choice in the late complete stenosis of the bile duct. Roux-en-Y hepatico-jejunostomy is the most common surgical procedure for the treatment of bile duct lesions and strictures. However, in high bile duct lesions, especially if the risk of anastomotic dehiscence is increased the Authors emphasize the Rodney-Smith technique for the reconstruction of the biliary tract.


Tankyrase-mediated β-catenin activity regulates vasopressin-induced AQP2 expression in kidney collecting duct mpkCCDc14 cells.
Aquaporin-2 (AQP2) mediates arginine vasopressin (AVP)-induced water reabsorption in the kidney collecting duct. AVP regulates AQP2 expression primarily via Gsα/cAMP/PKA signaling. Tankyrase, a member of the poly(ADP-ribose) polymerase family, is known to mediate Wnt/β-catenin signaling-induced gene expression. We examined whether tankyrase plays a role in AVP-induced AQP2 regulation via ADP-ribosylation of G protein-α (Gα) and/or β-catenin-mediated transcription of AQP2. RT-PCR and immunoblotting analysis revealed the mRNA and protein expression of tankyrase in mouse kidney and mouse collecting duct mpkCCDc14 cells. dDAVP-induced AQP2 upregulation was attenuated in mpkCCDc14 cells under the tankyrase inhibition by XAV939 treatment or small interfering (si) RNA knockdown. Fluorescence resonance energy transfer image analysis, however, revealed that XAV939 treatment did not affect dDAVP- or forskolin-induced PKA activation. Inhibition of tankyrase decreased dDAVP-induced phosphorylation of β-catenin (S552) and nuclear translocation of phospho-β-catenin. siRNA-mediated knockdown of β-catenin decreased forskolin-induced AQP2 transcription and dDAVP-induced AQP2 expression. Moreover, inhibition of phosphoinositide 3-kinase/Akt, which was associated with decreased nuclear translocation of β-catenin, diminished dDAVP-induced AQP2 upregulation, further indicating that β-catenin mediates AQP2 expression. Taken together, tankyrase plays a role in AVP-induced AQP2 regulation, which is likely via β-catenin-mediated transcription of AQP2, but not ADP-ribosylation of Gα. The results provide novel insights into vasopressin-mediated urine concentration and homeostasis of body water metabolism.


Children's and adults' eye movements and the extraction of number information from ambiguous and unambiguous markings.
Children learn to extract number information from utterances using a variety of language-specific cues, but in some languages this information is encoded in features that display high variability in their realization. How does this affect their acquisition process? In this paper, we present an eye tracking study comparing 3.5- to 7.5-year-old children and adults' use of number information on the verb and the determiner of the subject noun phrase in Chilean Spanish, a dialect of Spanish with variable realization of plural morphology in the noun phrase (due to phonological weakening) and categorical realization of number on the verb. Our results suggest that, while adults can determine whether the subject refers to a plurality or a singleton set based on the morphology of the verb alone, even 5- to 7.5-year-old children do not and, instead, require information from the noun phrase determiner to make a decision. Children younger than 5 years, instead, cannot use number information on the verb and on the determiner to make a decision. This supports Miller and Schmitt's (2010, 2012) hypothesis that number morphology is not always mapped into syntactic and semantic features by younger children in varieties of Spanish where number is subject to variation.


Smek promotes corticogenesis through regulating Mbd3's stability and Mbd3/NuRD complex recruitment to genes associated with neurogenesis.
The fate of neural progenitor cells (NPCs) during corticogenesis is determined by a complex interplay of genetic or epigenetic components, but the underlying mechanism is incompletely understood. Here, we demonstrate that Suppressor of Mek null (Smek) interact with methyl-CpG-binding domain 3 (Mbd3) and the complex plays a critical role in self-renewal and neuronal differentiation of NPCs. We found that Smek promotes Mbd3 polyubiquitylation and degradation, blocking recruitment of the repressive Mbd3/nucleosome remodeling and deacetylase (NuRD) complex at the neurogenesis-associated gene loci, and, as a consequence, increasing acetyl histone H3 activity and cortical neurogenesis. Furthermore, overexpression of Mbd3 significantly blocked neuronal differentiation of NPCs, and Mbd3 depletion rescued neurogenesis defects seen in Smek1/2 knockout mice. These results reveal a novel molecular mechanism underlying Smek/Mbd3/NuRD axis-mediated control of NPCs' self-renewal and neuronal differentiation during mammalian corticogenesis.


Large-scale collection and characterization of promoters of human and mouse genes.
We report the generation and initial characterization of a large-scale collection of sequences of putative promoter regions (PPRs) of human and mouse genes. Based on our unique collection of 400,225 and 580,209 human and mouse full-length cDNAs, we determined exact transcriptional start sites (TSSs). Using positional information of the TSSs, we could retrieve adjacent sequences as PPRs for 8,793 and 6,875 human and mouse genes, respectively. The positions of the PPRs were 4 kb upstream to previously reported 5'-ends of cDNAs on average, demonstrating that full-length cDNA information is indispensable for this purpose. Among those PPRs supported by experimentally validated TSSs, 3,324 could be paired as mutually homologous genes between human and mouse and were used for the comprehensive comparative studies. The sequence identities in the proximal regions of the TSSs were 45% on average, and 22,794 putative transcription factor binding sites that are conserved between human and mouse were identified. The data resource created in the present work and the results of the sequences' initial characterization should lay the firm foundation for deciphering the transcriptional modulations of human genes. All the data were deposited and made available through a database for comparative studies, DBTSS.


Case Report of Spontaneous Thyroid Hemorrhage Following LMA Insertion.
A case of thyroid hemorrhage following laryngeal mask airway (LMA) intubation is described. A 42-year-old female presented to the emergency department (ED) with left neck pain after same day surgery with LMA intubation and was found to have a thyroid hemorrhage on imaging. The patient was ultimately discharged in stable condition with no acute interventions performed in the ED or by otorhinolaryngology (ENT). Airway implications and possible interventions based on isolated case reports are reviewed and discussed. Thyroid hemorrhage, LMA complications.


A candidate pathway for a visual instructional signal to the barn owl's auditory system.
Many organisms use multimodal maps to generate coherent neuronal representations that allow adequate responses to stimuli that excite several sensory modalities. During ontogeny of these maps, one modality typically acts as the dominant system the other modalities are aligned to. A well studied model for the alignment of sensory maps is the calibration of the auditory space map by the visual system in the optic tectum of the barn owl. However, a projection from the optic tectum to the site of plasticity in the auditory pathway that could deliver an instructive signal has not been found so far. We have analyzed the development of the connectivity between the bimodal (visual and auditory) map of space in the barn owl's optic tectum and the auditory space map in the inferior colliculus with tracing methods and intracellular fills. Neurons in the tectal stratum griseum centrale were found to be suited to deliver an alignment signal from the visual midbrain to the auditory pathway. These neurons are presumably part of the efferent tectal projection pathway that mediates head saccades. The implications of a sensory alignment signal possibly being delivered by a (pre)motor command pathway are discussed.


[Adrenal glucocorticoid function and cytoplasmic dehydrogenase activity in the presnece of mechanical and toxic liver damage].
Against the background of low steroid metabolism in the liver there was noted some decrease in the rate of corticosterone synthesis by the adrenal gland sections, and also, a decrease in the dehydrogenase activity in the cytoplasm of adrenal cells in male rats 48 hours after partial hepatectomy, as compared to the sham-operated animals. These changes resulted from suppression of the central mechanisms of stress realization due to the lowered steroid metabolism. Intraperitoneal administration of carbon tetrachloride (0.1 ml per 100 g of body weight) at the same periods led to a significant intensification of the steroidogenesis in the adrenal tissue and to the activation of NAD-dependent dehydrogenases in the cytoplasm. The role of toxic injury of the glands in the changes of the functional state of the adrenal cortex cells is discussed.


The IPASC data format: A consensus data format for photoacoustic imaging.
Photoacoustic imaging (PAI) is an emerging modality that has shown promise for improving patient management in a range of applications. Unfortunately, the current lack of uniformity in PAI data formats compromises inter-user data exchange and comparison, which impedes: technological progress; effective research collaboration; and efforts to deliver multi-centre clinical trials. To overcome this challenge, the International Photoacoustic Standardisation Consortium (IPASC) has established a data format with a defined consensus metadata structure and developed an open-source software application programming interface (API) to enable conversion from proprietary file formats into the IPASC format. The format is based on Hierarchical Data Format 5 (HDF5) and designed to store photoacoustic raw time series data. Internal quality control mechanisms are included to ensure completeness and consistency of the converted data. By unifying the variety of proprietary data and metadata definitions into a consensus format, IPASC hopes to facilitate the exchange and comparison of PAI data.


Is Barrett's metaplasia the source of adenocarcinomas of the cardia?
To investigate the prevalence of Barrett's esophagus in patients with adenocarcinomas located at the gastroesophageal junction. A case series of patients who underwent esophagogastrectomy for adenocarcinoma was retrospectively reviewed. Tumors were grouped by location as esophageal, cardiac, and subcardiac, and the prevalence of specialized intestinal metaplasia in the histological specimens was determined. A university department of surgery that specializes in esophageal diseases. One hundred patients with adenocarcinoma of the esophagus, cardia, or proximal stomach. Cardiac adenocarcinomas were associated with Barrett's esophagus in 42% of the patients. Specialized intestinal metaplasia was identified in the histological sections from the resected specimen in 42% (13/31) of cardiac adenocarcinomas and in 79% (38/48) of esophageal adenocarcinomas but in only 5% (1/21) of subcardiac adenocarcinomas. The preoperative endoscopic biopsy results concurred with the final diagnosis of Barrett's esophagus in 33 of the 38 esophageal tumors, six of the 13 cardiac tumors, and the one subcardiac tumor but failed to detect specialized intestinal metaplasia in 54% (7/13) of cardiac tumors. Cardiac tumors were associated with shorter lengths of Barrett's mucosa than esophageal tumors (2.7 +/- 1.8 cm vs 7.4 +/- 3.4 cm, P < .01). The Barrett's metaplasia was dysplastic in 36 of the 38 esophageal tumors, 10 of the 13 cardiac tumors, but not in the subcardiac tumor. Adenocarcinomas located at the gastroesophageal junction were associated with Barrett's metaplasia in nearly one half of the patients. The length of the Barrett segment tends to be short and may be missed during endoscopy. The presence of high-grade dysplasia within Barrett's mucosa supports a barrett's origin for half of the adenocarcinomas arising at this location.


The development of gut microbiota in critically ill extremely low birth weight infants assessed with 16S rRNA gene based sequencing.
An increasing number of studies that are using high-throughput molecular methods are rapidly extending our knowledge of gut microbial colonization in preterm infants whose immaturity and requirement for extensive treatment may result in altered colonization process. We aimed to describe the profile of gut microbiota in 50 extremely low birth weight (<1200 g) critically ill infants at three different time points during the first two months of life by using 16S rRNA gene specific sequencing. Stool samples were collected at the age of one week, one month and two months. Bacterial community profiling was done using universal amplification of 16S rRNA gene and 454 pyrosequencing. The diversity of gut microbiota in preterm neonates in the first week of life was low but increased significantly over two months. The gut microbiota was dominated by facultative anaerobic bacteria (Staphylococcus spp. and Enterobacteriaceae) and lacked colonization with bacteria known to provide resistance against pathogens (Bacteroides, Bifidobacterium, and Lactobacillus) throughout the study. Colonization of Escherichia coli and uncultured Veillionella was positively correlated with maturity. Infants born to mothers with chorioamnionitis had significantly higher bacterial diversity than those without. High prevalence and abundance of potentially pathogenic Enterobacteriaceae and Staphylococcaceae with low prevalence and abundance of colonization resistance providing taxa bifidobacteria, Bacteroides and lactobacilli may lead to high infection risk via microbial translocation from the gut. Additionally, our data suggest that maternal chorioamnionitis may have an effect on the diversity of infants' gut microbiota; however, the mechanisms involved remain to be elucidated.


Disclosure documents and informed consent: the pros and cons.
This article critically examines the nature, scope, role and function as well as the pros and cons of disclosure documents as a means of procuring a patient's informed consent to proposed medical intervention. The conclusion reached is that, although the disadvantages of disclosure documents outnumber and outweigh their benefits, they should not be denied a role and function. Disclosure documents may be useful instruments for furnishing the basic information that a prudent patient would require to come to a decision whether to undergo or refuse the proposed medical intervention, but individual disclosure tailored to fit the actual circumstances of the particular patient can only be achieved by means of a disclosure conversation between doctor and patient. Hence, disclosure documents may serve as a supplement to but not as a substitute for a disclosure conversation.


Percutaneous nephrolithotomy in prone position in patients with spinal deformities.
The feasibility, safety and efficacy of percutaneous nephrolithotomy (PCNL) in patients with spinal deformities were evaluated and the results of a single centre experience were reported. Between July 1999 and December 2014, 16 patients with spinal deformities underwent PCNL. The anomalies included 5 cases with kyphoscoliosis, 4 with post-polio syndrome, 3 with osteogenesis imperfecta, 3 with myotonic dystrophy, and 1 with ankylosing spondylitis. All patients were preoperatively evaluated by an intravenous urogram and computerized tomography to assess the anatomy and appropriate access. The operative details, stone clearance rates, and complications were retrospectivelyanalyzed. A total of 16 standard PCNL procedures were performed on 16 renal-units. The mean age of the patients was 30.7 ± 17.2 (5-62) years, and the mean stone burden was 609.6 ± 526.9 (100-1800) mm(2). The mean operative and fluoroscopy times were 76.6 ± 35.1 (35-150) minutes and 12.5 ± 8.5 (3-34) minutes, respectively. At the end of the surgery, 13 (81.2%) of the patients were stone free. The overall success rate was 93.7% with the inclusion of 2 patients with clinically insignificant residual fragments (<3 mm). Complications (31.2%) included haemorrhage requiring a transfusion in 2 patients, prolonged urine leakage requiring double J catheter insertion in 1, infection in 1, and nephrectomy due to bleeding in 1. Mean hospitalization time was 4.6 ± 2.4 (3-13) days. PCNL is an effective, safe and minimally invasive procedure for the treatment of kidney stones in patients with spinal deformities, and it can be performed with low morbidity and high success rates. To achieve better results and minimizing the risk factors, systematic and anatomic evaluations for anaesthesia and operative planning are crucial before surgery.


Ontogenetic niche shifts in a locally endangered tree species (Olea europaea subsp. cuspidata) in a disturbed forest in Northern Ethiopia: Implications for conservation.
Understanding the responses of different ontogenetic stages to environmental and human disturbance factors is essential for developing efficient conservation strategies for endangered plant species. We examined how three ontogenetic stages of a locally endangered tree species, Olea europaea subsp. cuspidata, responded to environmental factors and human disturbance in Hugumburda dry Afromontane forest in Ethiopia. We counted individual seedlings, saplings and adults of O. europaea in 70 20 × 20 m quadrats over ca. 2.8 ha, and measured biotic (woody species richness, canopy cover, aboveground tree biomass, herbaceous cover), abiotic (soil and topographic variables), and human disturbance factors (logging and tracks). To detect ontogenetic niche shifts, we compared observed vs. simulated locations of trees in the three life stages and how they related to the environmental and human disturbance factors. We found that the population structure of O. europaea showed generally low recruitment, with few seedlings per hectare compared with the abundance of saplings and adults. The probability of finding O. europaea individuals was influenced by biotic (woody species richness) and abiotic (soil depth, slope) environmental conditions and human disturbance (logging intensity), but the direction, strength and shape of the relationships differed between seedling, sapling and adult life stages, indicating ontogenetic niche shifts. All life stages showed a positive relationship with elevation. The observed environmental niches of the different lifestages of O. europaea, and their association with human disturbance levels, should be considered when conservation strategies are developed for this species. Human disturbance in terms of logging decreases the abundance of saplings, but may facilitate emerging seedlings through creation of gaps with improved light conditions. Recruitment is, however, very low in the study area, and seedlings should be protected from browsing to enhance survival. Woody species richness in general should be conserved to optimize conditions also for O. europaea saplings.


CRISPR Typing Increases the Discriminatory Power of Streptococcus agalactiae Typing Methods.
We explored the relevance of a Clustered regularly interspaced short palindromic repeats (CRISPR)-based genotyping tool for Streptococcus agalactiae typing and we compared this method to current molecular methods [multi locus sequence typing (MLST) and capsular typing]. To this effect, we developed two CRISPR marker schemes (using 94 or 25 markers, respectively). Among the 255 S. agalactiae isolates tested, 229 CRISPR profiles were obtained. The 94 and 25 markers made it possible to efficiently separate isolates with a high diversity index (0.9947 and 0.9267, respectively), highlighting a high discriminatory power, superior to that of both capsular typing and MLST (diversity index of 0.9017 for MLST). This method has the advantage of being correlated with MLST [through analysis of the terminal direct repeat (TDR) and ancestral spacers] and to possess a high discriminatory power (through analysis of the leader-end spacers recently acquired, which are the witnesses of genetic mobile elements encountered by the bacteria). Furthermore, this "one-shot" approach presents the benefit of much-reduced time and cost in comparison with MLST. On the basis of these data, we propose that this method could become a reference method for group B Streptococcus (GBS) typing.


Moxifloxacin in Pediatric Patients With Complicated Intra-abdominal Infections: Results of the MOXIPEDIA Randomized Controlled Study.
This study was designed to evaluate primarily the safety and also the efficacy of moxifloxacin (MXF) in children with complicated intra-abdominal infections (cIAIs). In this multicenter, randomized, double-blind, controlled study, 451 pediatric patients aged 3 months to 17 years with cIAIs were treated with intravenous/oral MXF (N = 301) or comparator (COMP, intravenous ertapenem followed by oral amoxicillin/clavulanate; N = 150) for 5 to 14 days. Doses of MXF were selected based on the results of a Phase 1 study in pediatric patients (NCT01049022). The primary endpoint was safety, with particular focus on cardiac and musculoskeletal safety; clinical and bacteriologic efficacy at test of cure was also investigated. The proportion of patients with adverse events (AEs) was comparable between the 2 treatment arms (MXF: 58.1% and COMP: 54.7%). The incidence of drug-related AEs was higher in the MXF arm than in the COMP arm (14.3% and 6.7%, respectively). No cases of QTc interval prolongation-related morbidity or mortality were observed. The proportion of patients with musculoskeletal AEs was comparable between treatment arms; no drug-related events were reported. Clinical cure rates were 84.6% and 95.5% in the MXF and COMP arms, respectively, in patients with confirmed pathogen(s) at baseline. MXF treatment was well tolerated in children with cIAIs. However, a lower clinical cure rate was observed with MXF treatment compared with COMP. This study does not support a recommendation of MXF for children with cIAIs when alternative more efficacious antibiotics with better safety profile are available.


An evaluation of the Abaxis VSPro for the measurement of equine plasma fibrinogen concentrations.
Accurate measurement of plasma fibrinogen concentrations is an important tool for assessment of horses with inflammatory diseases. To determine the precision and accuracy of a benchtop instrument using both fresh and frozen equine plasma by comparing the plasma fibrinogen concentration measured by a benchtop instrument to 2 separate laboratory standard methods (ACL 100 and STA Compact) for fibrinogen measurement. Accuracy and precision of the VSPro was evaluated using both human fibrinogen standards and samples from horses. Fifty frozen samples from horses with gastrointestinal disease had the fibrinogen concentration measured using the ACL 1000 and the VSPro. Fifty fresh samples were collected from hospitalised horses and fibrinogen concentration was measured using the STA Compact coagulation machine and the VSPro. Correlations for measurements were performed, as well as Bland-Altman analysis. Coefficients of variability for the VSPro ranged from 7% to 15%. The VSPro fibrinogen values were well correlated to both the ACL 1000 (r = 0.94, P<0.001) and the STA Compact measurements (r = 0.926, P<0.001). Bland-Altman analysis showed a mean bias of -0.83 g/l (95% confidence interval -2.03-0.324 g/l) for the ACL 1000 and a mean bias of -0.024 g/l (95% confidence interval -1.434-1.386 g/l) for the STA Compact. The VSPro appears to have adequate accuracy and precision for clinical measurement of plasma fibrinogen concentrations. The VSPro provides a measurement of equine plasma fibrinogen concentration using a benchtop instrument with a rapid test time that has comparable accuracy to the fibrinogen concentration obtained from reference laboratories.


Microbiome-based body fluid identification of samples exposed to indoor conditions.
In the forensic reconstruction of crime scene activities, the identification of biological traces and their bodily origin are valuable evidence that can be presented in court. While several presumptive and confirmatory tests are currently available, the limitations in specificity and sensitivity have instigated a search for alternative methods. Bacterial markers have been proposed as a novel approach for forensic body fluid/tissue identification. Bacteria are not only ubiquitous throughout the human body, but also, as shown by recent microbiome sequencing studies of the 16S rRNA gene, bacterial community structures are distinct across body sites. Traces and stains at crime scenes are, however, often exposed to the environment outside the human body for variable periods of time before laboratory processing. Thus, it is not clear whether exposed samples continue to harbor microbial signatures characteristic of their body site of origin. In this proof-of-concept study we collected samples from six different body sites: saliva, skin, peripheral blood, vaginal fluid, menstrual blood and semen. We exposed a subset of these samples to indoor conditions for 30 days while the remaining samples were processed directly after extraction. Our analyses of 16S rRNA gene sequence data for a total of 46 control and exposed samples show that both types of samples group by body site, although a few outliers are observed. Based on our results, vaginal and menstrual samples share their microbial signatures, and cannot be distinguished using bacterial markers. Overall, our findings indicate that bacterial markers are a promising avenue for forensic body fluid/tissue identification.


[Comparative action of melatonin and bilobil on light perception and psychophysiological indices in patients with cerebral trauma anamnesis].
Pineal hormone melatonin (melaxen preparation) and a typical cognitive enhancer Ginkgo biloba (bilobil preparation) decreased the retinal light sensitivity thresholds upon two-week administration in young (aged 19-23) patients with cerebral trauma anamnesis. This effect was more significant in evening than in the morning hours. Both drugs also attenuated the reactive anxiety (bilobil was slightly superior to melaxen in this respect) and insignificantly increased the volume of acoustic memory. The observed similarity of the psychotropic effect of two preparations allows melatonin to be classified into nootropic agents.


In vitro and in vivo pharmacokinetic-pharmacodynamic relationships for the trisubstituted aminopurine cyclin-dependent kinase inhibitors olomoucine, bohemine and CYC202.
To investigate pharmacokinetic-pharmacodynamic relationships for the trisubstituted aminopurine cyclin-dependent kinase inhibitors olomoucine, bohemine, and CYC202 (R-roscovitine; seliciclib) in the HCT116 human colon carcinoma model. The in vitro activity of the agents was determined in a human tumor panel using the sulforhodamine B assay. The concentration and time dependence was established in HCT116 cells. Molecular biomarkers, including RB phosphorylation and cyclin expression, were assessed by Western blotting. Pharmacokinetic properties were characterized in mice following analysis by liquid chromatography-tandem mass spectrometry. Based on these studies, a dosing regimen was developed for CYC202 that allowed therapeutic exposures in the HCT116 tumor xenograft. The antitumor potency of the agents in vitro was in the order olomoucine (IC50, 56 micromol/L) < bohemine (IC50, 27 micromol/L) < CYC202 (IC50, 15 micromol/L), corresponding to their activities as cyclin-dependent kinase inhibitors. Antitumor activity increased with exposure time up to 16 hours. The agents caused inhibition of RB and RNA polymerase II phosphorylation and depletion of cyclins. They exhibited relatively rapid clearance following administration to mice. CYC202 displayed the slowest clearance from plasma and the highest tumor uptake, with oral bioavailability of 86%. Oral dosing of CYC202 gave active concentrations in the tumor, modulation of pharmacodynamic markers, and inhibition of tumor growth. CYC202 showed therapeutic activity on human cancer cell lines in vitro and on xenografts. Pharmacodynamic markers are altered in vitro and in vivo, consistent with the inhibition of cyclin-dependent kinases. Such markers may be potentially useful in the clinical development of CYC202 and other cyclin-dependent kinase inhibitors.