Independent Validation of Early-Stage Non-Small Cell Lung Cancer Prognostic Scores Incorporating Epigenetic and Transcriptional Biomarkers With Gene-Gene Interactions and Main Effects.
DNA methylation and gene expression are promising biomarkers of various cancers, including non-small cell lung cancer (NSCLC). Besides the main effects of biomarkers, the progression of complex diseases is also influenced by gene-gene (G×G) interactions. Would screening the functional capacity of biomarkers on the basis of main effects or interactions, using multiomics data, improve the accuracy of cancer prognosis? Biomarker screening and model validation were used to construct and validate a prognostic prediction model. NSCLC prognosis-associated biomarkers were identified on the basis of either their main effects or interactions with two types of omics data. A prognostic score incorporating epigenetic and transcriptional biomarkers, as well as clinical information, was independently validated. Twenty-six pairs of biomarkers with G×G interactions and two biomarkers with main effects were significantly associated with NSCLC survival. Compared with a model using clinical information only, the accuracy of the epigenetic and transcriptional biomarker-based prognostic model, measured by area under the receiver operating characteristic curve (AUC), increased by 35.38% (95% CI, 27.09%-42.17%; P = 5.10 × 10-17) and 34.85% (95% CI, 26.33%-41.87%; P = 2.52 × 10-18) for 3- and 5-year survival, respectively, which exhibited a superior predictive ability for NSCLC survival (AUC3 year, 0.88 [95% CI, 0.83-0.93]; and AUC5 year, 0.89 [95% CI, 0.83-0.93]) in an independent Cancer Genome Atlas population. G×G interactions contributed a 65.2% and 91.3% increase in prediction accuracy for 3- and 5-year survival, respectively. The integration of epigenetic and transcriptional biomarkers with main effects and G×G interactions significantly improves the accuracy of prognostic prediction of early-stage NSCLC survival.


Issues raised by respiratory therapy arrangements.
In large part due to the foregoing issues, Medicare program officials have focused on respiratory therapy as an area with great potential for abuse, and may well introduce significant reforms in the near future. Accordingly, any contractual arrangements for respiratory therapy programs of the type discussed above should be carefully reviewed for compliance with Medicare requirements and for a realistic assessment of the parties' potential exposure to liability under the anti-kickback statute. At a minimum, these agreements should provide for short "without cause" termination provisions, or include a so-called "jeopardy" provision that permits the parties to renegotiate or terminate their contract if significant changes occur in, or if the current arrangement is found or threatened to be found to violate, applicable law.


Efflux pumps are involved in the defense of Gram-negative bacteria against the natural products isobavachalcone and diospyrone.
The activities of two naturally occurring compounds, isobavachalcone and diospyrone, against documented strains and multidrug-resistant (MDR) Gram-negative bacterial isolates were evaluated. The results indicated that the two compounds exhibited intrinsic antibacterial activity against several Gram-negative bacteria, and their activities were significantly improved in the presence of an efflux pump inhibitor (MIC values decreased to below 10 microg/ml). In addition, the activities of isobavachalcone and diospyrone against various strains exhibiting deletions of the major efflux pump components (AcrAB, TolC) were significantly increased. The overall results indicate that isobavachalcone and diospyrone could be candidates for the development of new drugs against MDR strains and that their use in combination with efflux pump inhibitors reinforces their activity.


Parthenolide Destabilizes Microtubules by Covalently Modifying Tubulin.
Detyrosination of the α-tubulin C-terminal tail is a post-translational modification (PTM) of microtubules that is key for many biological processes.1 Although detyrosination is the oldest known microtubule PTM,2-7 the carboxypeptidase responsible for this modification, VASH1/2-SVBP, was identified only 3 years ago,8,9 precluding genetic approaches to prevent detyrosination. Studies examining the cellular functions of detyrosination have therefore relied on a natural product, parthenolide, which is widely believed to block detyrosination of α-tubulin in cells, presumably by inhibiting the activity of the relevant carboxypeptidase(s).10 Parthenolide is a sesquiterpene lactone that forms covalent linkages predominantly with exposed thiol groups; e.g., on cysteine residues.11-13 Using mass spectrometry, we show that parthenolide forms adducts on both cysteine and histidine residues on tubulin itself, in vitro and in cells. Parthenolide causes tubulin protein aggregation and prevents the formation of microtubules. In contrast to epoY, an epoxide inhibitor of VASH1/2-SVBP,9 parthenolide does not block VASH1-SVBP activity in vitro. Lastly, we show that epoY is an efficacious inhibitor of microtubule detyrosination in cells, providing an alternative chemical means to block detyrosination. Collectively, our work supports the notion that parthenolide is a promiscuous inhibitor of many cellular processes and suggests that its ability to block detyrosination may be an indirect consequence of reducing the polymerization-competent pool of tubulin in cells.


Preliminary assessment of the effect of more informative (structured) abstracts on citation retrieval from MEDLINE.
To compare the recall and precision of citation retrieval from MEDLINE when searching in journals containing traditional abstracts with journals containing abstracts prepared according to the guidelines of more informative abstracts (MIAs) for clinical journal articles. Analytic survey of operating characteristics of MeSH terms and textwords selected to detect studies meeting basic methodological criteria for direct clinical use in general adult medicine in priority journals with and without MIAs. The sensitivity, specificity, and precision of search terms were determined by comparison with a manual review of all articles (the gold standard) in 10 internal and general medicine journals for 1986 and 1991. The performance of each search term was compared before and after the implementation of MIAs and in two journal sets with and without MIAs in 1991. The sensitivity of some terms increased with the use of MIAs and also increased by the same amount in journals with usual abstracts in 1991 compared to 1986. The use of MIAs had the greatest impact in the areas of etiology and prognosis. Many MeSH and textwords had comparable performance before and after the implementation of MIAs and between journals with and without MIAs in 1991. More informative abstracts improved the retrieval properties of some, but not all, textwords and medical subject headings. Further work may be needed to enhance the effects of MIAs on bibliographic retrieval.


Hypolipidaemic effect of Casearia esculenta root extracts in streptozotocin-induced diabetic rats.
The hypolipidaemic effect of an aqueous extract of Casearia esculenta root, an indigenous antidiabetic medicine popularly used in rural South India was investigated. Administration of the extract of C. esculenta (200 and 300 mg/kg body wt.) for 45 days resulted in significant reduction in serum and tissue cholesterol, phospholipids, free fatty acids and triglycerides in streptozotocin (STZ) diabetic rats. In addition to that, significant (p < 0.05) decrease in high density lipoprotein (HDL) whereas significant increase (p < 0.05) in low density lipoprotein (LDL) and very low density lipoprotein (VLDL) were observed in STZ diabetic rats, which was normalized after 45 days of C. esculenta root extract treatment. The root extract at dose of 300 mg/kg body wt. showed much significant hypolipidaemic effects than the dose of 200 mg/kg body weight.


Difference in Functional Performance on the Upper-Quarter Y-Balance Test Between High School Baseball Players and Wrestlers.
The Upper Quarter Y-Balance Test (YBT-UQ) is a unique movement test where individuals perform at the limits of their stability, requiring the coordination of balance, proprioception, range of motion, and stabilization. It is not yet clear if performance on the YBT-UQ differs between sports with dissimilar emphasis on upper-extremity performance. To compare performance on the YBT-UQ between wrestlers, whose sport requires some degree of closed-chain activity, and baseball players, whose sport is primarily open kinetic chain in nature. Cross-sectional. High school preparticipation physical assessment. 24 healthy high school male wrestlers (mean age 16.12 ± 1.24 y) and 24 healthy high school male baseball players (mean age 15.79 ± 1.25 y). All subjects performed the YBT-UQ, which requires reaching in 3 directions while maintaining a push-up position. The variables of interest include the maximum reach in each direction, as well as the composite score. In addition, asymmetries between limbs for each reach direction were compared. Wrestlers performed significantly better than baseball players in the medial direction, inferolateral direction, and in composite scores. In the medial direction, wrestlers exhibited greater scores (P < .01) on both left and right limbs, 10.5 ± 10.2%LL and 9.95 ± 10.2%LL, respectively. Significant differences (P < .01) were also observed in the inferolateral direction, with a difference of 11.3 ± 12.0%LL on the left and 8.7 ± 11.0%LL on the right. Composite scores were higher (P < .01) for the wrestlers, with a difference of 7.0% on the left and 7.1% on the right. This study suggests that wrestlers perform better on the YBT-UQ than baseball players. The findings may suggest sport-specific normative data for the YBT-UQ in high school athletes.


Effects of binding policy and other factors on the availability of journal issues.
Random samples of journal issues, derived from a test set of 106 biomedical journal titles, were checked against the shelves of the Woodward Biomedical Library under circulating and noncirculating conditions. As well as circulation, binding procedures and duplication of sets were seen to affect availability. Indications are that the practice of binding journals upon completion of the volume may remove them from use at a time of peak demand. The usefulness of second sets of journals appeared to decline rapidly within a few years after publication. A change from circulating to noncirculating conditions may have increased availability certain journals by discouraging use.


A prospective study of the immune system activation biomarker neopterin and colorectal cancer risk.
Neopterin may be relevant for colorectal cancer (CRC) development, as a biomarker of cellular immune activity exerting pleiotropic effects on cellular ageing, oxidative stress, and inflammation. So far, the association between prediagnostic neopterin and colon and rectal cancer risk has not been evaluated in human populations. A nested case-control study was conducted within the European Prospective Investigation into Cancer and Nutrition cohort using data on plasma concentrations of total neopterin (T-N, sum of neopterin and 7,8-dihydroneopterin) in 830 incident CRC case patients (561 colon and 269 rectal) matched within risk sets to 830 control participants. A subsequent replication study used data from the Hordaland Health Study, where 173 CRC case patients have been diagnosed among 6594 healthy participants over 12 years of follow-up. After multivariable adjustment for a priori chosen CRC risk factors, a "U-shaped" association of T-N with CRC was revealed. Compared with the second quintile of the T-N distribution, the relative risks for the first, third, fourth, and fifth quintiles were 2.37 (95% CI = 1.66 to 3.39), 1.24 (95% CI = 0.87 to 1.77), 1.55 (95% CI = 1.08 to 2.22), and 2.31 (95% CI = 1.63 to 3.27), respectively. Replication of these associations within the Hordaland Health Study yielded similar results. No differences have been observed when the associations were explored by colon and rectal cancer site (two-sided P difference = .87) and after excluding case patients diagnosed within the first four follow-up years. These novel findings provide evidence of the role of both suppressed and activated cell-mediated immunity as reflected by prediagnostic T-N concentrations in the development of CRC.


[Penetrating wounds of abdominal cavity in children].
An analysis of treatment of 26 children with penetrating wounds of the abdominal cavity has been made. Extensive laparotomy and careful revision of internal organs is shown to be necessary on the establishing of the penetrating character of the wound. The presence of shock is not a contraindication to operation. Modern anesthesiological methods and intensive treatment in the postoperative period improve results of the treatment.


Argyrophilic grain disease as a neurodegenerative substrate in late-onset schizophrenia and delusional disorders.
To study the relationship between neurodegenerative diseases including argyrophilic grain disease (AGD) and late-onset schizophrenia and delusional disorders (LOSD; onset ≥40 years of age), we pathologically examined 23 patients with LOSD, 71 age-matched normal controls, and 22 psychiatric disease controls (11 depression, six personality disorder, two bipolar disorders, and three neurotic disorders cases). In all LOSD cases (compared to age-matched normal controls), the frequencies of Lewy body disease (LBD), AGD, and corticobasal degeneration (CBD) were 26.1 % (11.3 %), 21.7 % (8.5 %), and 4.3 % (0.0 %), respectively. There was no case of pure Alzheimer's disease (AD). The total frequency of LBD, AGD, and CBD was significantly higher in LOSD cases than in normal controls. Argyrophilic grains were significantly more severe in LOSD than in controls, but were almost completely restricted to the limbic system and adjacent temporal cortex. In LOSD patients whose onset occurred at ≥65 years of age (versus age-matched normal controls), the frequencies of LBD and AGD were 36.4 % (19.4 %) and 36.4 % (8.3 %), respectively, and AGD was significantly more frequent in LOSD patients than in normal controls. In LOSD patients whose onset occurred at <65 years of age, the frequencies of LBD, AGD, and CBD were 16.7, 8.3, and 8.3 %, comparable to those of age-matched normal controls (10.2, 5.1, and 0.0 %). In all psychiatric cases, delusion was significantly more frequent in AGD cases than in cases bearing minimal AD pathology alone. Given these findings, LOSD patients may have heterogeneous pathological backgrounds, and AGD may be associated with the occurrence of LOSD especially after 65 years of age.


Evidence-based treatment of chronic heart failure.
The past two decades have seen a knowledge explosion in the field of cardiovascular diseases, in general, and in the understanding of chronic heart failure (HF) as a complex neurohumoral syndrome in particular. A new staging system for chronic HF has been developed within the last decade to facilitate the evidence-based prescription of medications and medical devices for each of its four stages. The burden of care for patients with chronic HF is substantially provided in primary care settings. Primary care physicians need to understand the underlying pathophysiology of chronic HF, the elements of its evaluation and treatment by stage, and when referral is necessary.


Systematic review of interventions to improve the psychological well-being of general practitioners.
The health of doctors who work in primary care is threatened by workforce and workload issues. There is a need to find and appraise ways in which to protect their mental health, including how to achieve the broader, positive outcome of well-being. Our primary outcome was to evaluate systematically the research evidence regarding the effectiveness of interventions designed to improve General Practitioner (GP) well-being across two continua; psychopathology (mental ill-health focus) and 'languishing to flourishing' (positive mental health focus). In addition we explored the extent to which developments in well-being research may be integrated within existing approaches to design an intervention that will promote mental health and prevent mental illness among these doctors. Medline, Embase, Cinahl, PsychINFO, Cochrane Register of Trials and Web of Science were searched from inception to January 2015 for studies where General Practitioners and synonyms were the primary participants. Eligible interventions included mental ill-health prevention strategies (e.g. promotion of early help-seeking) and mental health promotion programmes (e.g. targeting the development of protective factors at individual and organizational levels). A control group was the minimum design requirement for study inclusion and primary outcomes had to be assessed by validated measures of well-being or mental ill-health. Titles and abstracts were assessed independently by two reviewers with 99% agreement and full papers were appraised critically using validated tools. Only four studies (with a total of 997 GPs) from 5392 titles met inclusion criteria. The studies reported statistically significant improvement in self-reported mental ill-health. Two interventions used cognitive-behavioural techniques, one was mindfulness-based and one fed-back GHQ scores and self-help information. There is an urgent need for high quality, controlled studies in GP well-being. Research on improving GP well-being is limited by focusing mainly on stressors and not giving systematic attention to the development of positive mental health.


Thromboembolism and pancreatic cancer.
Venous thromboembolism (VTE) is a frequent event in the clinical course of patients with exocrine pancreatic cancer; studies have been designed to evaluate the role of prophylactic anticoagulation in this ominous disease. Searching for the molecular basis of thrombosis in cancer, Bozkurt et al. present in the Abstract #e22049 the result of their investigation on the frequency of inherited and carcinogenesis-acquired proteins in oncologic patients with and without venous thromboembolism. From the bedside, Muñoz Martin et al. present in the Abstract #e15187 their work on the incidence of venous thromboembolism in patients with exocrine pancreatic cancer and the role of the established Khorana score in predicting symptomatic and incidental venous thromboembolism. At last, Cella et al. in the Abstract #e20625 expand the predictor landscape from the Khorana score to other risk factors for venous thromboembolism, refining the selection of oncologic patients who can benefit from prophylactic anticoagulation.


Ginsenosides Act As Positive Modulators of P2X4 Receptors.
We investigated the selectivity of protopanaxadiol ginsenosides from Panax ginseng acting as positive allosteric modulators on P2X receptors. ATP-induced responses were measured in stable cell lines overexpressing human P2X4 using a YOPRO-1 dye uptake assay, intracellular calcium measurements, and whole-cell patch-clamp recordings. Ginsenosides CK and Rd were demonstrated to enhance ATP responses at P2X4 by ∼twofold, similar to potentiation by the known positive modulator ivermectin. Investigations into the role of P2X4 in mediating a cytotoxic effect showed that only P2X7 expression in HEK-293 cells induces cell death in response to high concentrations of ATP, and that ginsenosides can enhance this process. Generation of a P2X7-deficient clone of BV-2 microglial cells using CRISPR/Cas9 gene editing enabled an investigation of endogenous P2X4 in a microglial cell line. Compared with parental BV-2 cells, P2X7-deficient BV-2 cells showed minor potentiation of ATP responses by ginsenosides, and insensitivity to ATP- or ATP+ ginsenoside-induced cell death, indicating a primary role for P2X7 receptors in both of these effects. Computational docking to a homology model of human P2X4, based on the open state of zfP2X4, yielded evidence of a putative ginsenoside binding site in P2X4 in the central vestibule region of the large ectodomain.


Tumor immunotargeting using innovative radionuclides.
This paper reviews some aspects and recent developments in the use of antibodies to target radionuclides for tumor imaging and therapy. While radiolabeled antibodies have been considered for many years in this context, only a few have reached the level of routine clinical use. However, alternative radionuclides, with more appropriate physical properties, such as lutetium-177 or copper-67, as well as alpha-emitting radionuclides, including astatine-211, bismuth-213, actinium-225, and others are currently reviving hopes in cancer treatments, both in hematological diseases and solid tumors. At the same time, PET imaging, with short-lived radionuclides, such as gallium-68, fluorine-18 or copper-64, or long half-life ones, particularly iodine-124 and zirconium-89 now offers new perspectives in immuno-specific phenotype tumor imaging. New antibody analogues and pretargeting strategies have also considerably improved the performances of tumor immunotargeting and completely renewed the interest in these approaches for imaging and therapy by providing theranostics, companion diagnostics and news tools to make personalized medicine a reality.


Complete pathway elucidation and heterologous reconstitution of (+)-nootkatone biosynthesis from Alpinia oxyphylla.
(+)-Nootkatone is a natural sesquiterpene ketone widely used in food, cosmetics, pharmaceuticals, and agriculture. It is also regarded as one of the most valuable terpenes used commercially. However, plants contain trace amounts of (+)-nootkatone, and extraction from plants is insufficient to meet market demand. Alpinia oxyphylla is a well-known medicinal plant in China, and (+)-nootkatone is one of the main components within the fruits. By transcriptome mining and functional screening using a precursor-providing yeast chassis, the complete (+)-nootkatone biosynthetic pathway in Alpinia oxyphylla was identified. A (+)-valencene synthase (AoVS) was identified as a novel monocot-derived valencene synthase; three (+)-valencene oxidases AoCYP6 (CYP71BB2), AoCYP9 (CYP71CX8), and AoCYP18 (CYP701A170) were identified by constructing a valencene-providing yeast strain. With further characterisation of a cytochrome P450 reductase (AoCPR1) and three dehydrogenases (AoSDR1/2/3), we successfully reconstructed the (+)-nootkatone biosynthetic pathway in Saccharomyces cerevisiae, representing a basis for its biotechnological production. Identifying the biosynthetic pathway of (+)-nootkatone in A. oxyphylla unravelled the molecular mechanism underlying its formation in planta and also supported the bioengineering production of (+)-nootkatone. The highly efficient yeast chassis screening method could be used to elucidate the complete biosynthetic pathway of other valuable plant natural products in future.


Carbohydrate intake and food sources of junior triathletes during a moderate and an intensive training period.
The study was conducted to investigate the quantity and the main food sources of carbohydrate (CHO) intake of junior elite triathletes during a short-term moderate (MOD; 12 km swimming, 100 km cycling, 30 km running per wk) and intensive training period (INT; 23 km swimming, 200 km cycling, 45 km running per wk). Self-reported dietary-intake data accompanied by training protocols of 7 male triathletes (18.1 ± 2.4 yr, 20.9 ± 1.4 kg/m(2)) were collected on 7 consecutive days during both training periods in the same competitive season. Total energy and CHO intake were calculated based on the German Food Database. A paired t test was applied to test for differences between the training phases (α = .05). CHO intake was slightly higher in INT than in MOD (9.0 ± 1.6 g · kg(-1) · d(-1) vs. 7.8 ± 1.6 g · kg(-1) · d(-1); p = .041). Additional CHO in INT was mainly ingested during breakfast (115 ± 37 g in MOD vs. 175 ± 23 g in INT; p = .002) and provided by beverages (280.5 ± 97.3 g/d vs. 174.0 ± 58.3 g/d CHO; p = .112). Altogether, main meals provided approximately two thirds of the total CHO intake. Pre- and postexercise snacks additionally supplied remarkable amounts of CHO (198.3 ± 84.3 g/d in INT vs. 185.9 ± 112 g/d CHO in MOD; p = .231). In conclusion, male German junior triathletes consume CHO in amounts currently recommended for endurance athletes during moderate to intensive training periods. Main meals provide the majority of CHO and should therefore not be skipped. CHO-containing beverages, as well as pre- and postexercise snacks, may provide a substantial amount of CHO intake in training periods with high CHO requirements.


Early isolated optic neuropathy caused by cyclosporine.
To examine, for the first time, whether cyclosporine intake has an early isolated effect on the optic nerve. This observational case series consisted of 192 eyes of 98 patients treated with cyclosporine. Patient age and duration and dosage of cyclosporine were recorded, and visual acuity, optic nerve function, visual fields, and visual evoked potential (VEP) were tested. Fundus examination was also performed. Patients with glaucoma, vascular retinopathies, and deep amblyopia were excluded. Mean patient age was 46 years, average duration of treatment was 6 years, and median dosage of cyclosporine was 200 mg daily. VEP was tested in 73 patients (142 eyes) and yielded a delayed P100 wave in 9 (12.32%) (14 eyes). Among these 9 patients, abnormal findings were also noted on the Ishihara colour test in 42.86% of the eyes, and on the visual field test in 64.3% of the eyes. Abnormal VEP showed a significant correlation (p < 0.05) with older age (> 46 years) and a non-significant correlation with longer duration of treatment. Higher abnormal VEP potential was not correlated with higher cyclosporine dose, and there was no correlation between abnormal VEP and blood level of cyclosporine. Optic neuropathy was significantly associated with older age in cyclosporine-treated patients. A correlation between optic neuropathy with longer duration of cyclosporine treatment was noted but was not statistically significant. We suggest that tests of optic nerve function, including VEP, be a part of the follow-up of patients receiving cyclosporine.


Cellular localization and trafficking of vascular adhesion protein-1 as revealed by an N-terminal GFP fusion protein.
Recent studies of vascular adhesion protein-1 (VAP-1) have greatly advanced our understanding of the important role this protein plays in the establishment and progression of inflammatory disease. To facilitate more detailed studies on the function of VAP-1, we developed a GFP-fusion protein that enabled us to monitor the trafficking of the protein in three selected cell types: hepatic sinusoidal endothelial cells, liver myofibroblasts and an hepatic stellate cell line (LX-2). The fusion protein was detected as punctate cytoplasmic GFP staining, but was present only at low levels at the cell surface in all cell types studied. The subcellular distribution of the protein was not altered in a catalytically inactive mutant form of the protein (Tyr471Phe) or in the presence of exogenous VAP-1 substrate (methylamine) or inhibitor (semicarbazide). The GFP-VAP-1 protein was localized to the Golgi apparatus (GM-130), endoplasmic reticulum (GRP94) and early endosomes (EEA-1). Additional staining for VAP-1 revealed that the overexpressed protein was also present in vesicles that were negative for GFP fluorescent signal and did not express EEA-1. We propose that these vesicles are responsible for recycling the fusion protein and that the fluorescence of the GFP moiety is quenched at the low pH within these vesicles. This feature of the protein makes it well suited for live cell imaging studies where we wish to track protein that is being actively trafficked within the cell in preference to that which is being recycled.


An Interactive Web Portal for Tracking Oncology Patient Physical Activity and Symptoms: Prospective Cohort Study.
Physical activity levels typically decline during cancer treatment and often do not return to prediagnosis or minimum recommended levels. Interventions to promote physical activity are needed. Support through the use of digital health tools may be helpful in this situation. The goal of the research was to evaluate the feasibility, usability, and acceptability of an interactive Web portal developed to support patients with cancer to increase daily physical activity levels. A Web portal for supportive cancer care which was developed to act as a patient-clinician information and coaching tool focused on integrating wearable device data and remote symptom reporting. Patients currently receiving or who had completed intensive anticancer therapy were recruited to 3 cohorts. All cohorts were given access to the Web portal and an activity monitor over a 10-week period. Cohort 2 received additional summative messaging, and cohort 3 received personalized coaching messaging. Qualitative semistructured interviews were completed following the intervention. The primary outcome was feasibility of the use of the portal assessed as both the number of log-ins to the portal to record symptoms and the completion of post-program questionnaires. Of the 49 people were recruited, 40 completed the intervention. Engagement increased with more health professional contact and was highest in cohort 3. The intervention was found to be acceptable by participants. The portal was feasible for use by people with a history of cancer. Further research is needed to determine optimal coaching methods.


Associations Between Depression/Anxiety and Headache Frequency in Migraineurs: A Cross-Sectional Study.
While migraines have been associated with emotional disturbances, it remains unknown whether the intensity of emotional expression is directly related to migraine frequency. The present study investigated depression/anxiety among migraineurs. This cross-sectional study included 588 clinical outpatients in Taiwan. Migraines were stratified by attack frequency, with and without auras, and with well-controlled confounding variables. Demographic and clinical data, including sleep characteristics, were collected. Multivariable linear regressions were employed to examine whether migraine frequency (1-4 headache days per month, 5-8 headache days per month, 9-14 headache days per month, or >14 headache days per month) was associated with depression/anxiety symptoms, as indicated by the Beck's Depression Inventory (BDI) and Hospital Anxiety and Depression Subscales (HADS). BDI total scores were highest in patients with chronic migraines (mean ± SD: 13.2 ± 8.5), followed by those with high frequency (12.1 ± 8.5), medium frequency (10.6 ± 8.0), low frequency (9.1 ± 7.1), and lowest in nonmigraine controls (6.6 ± 5.9), with a significant trend in frequency (P trend < .001); similar results were obtained for HADS scores. BDI and HADS scores were independently related to high-frequency episodic and chronic migraine frequency and to poor sleep quality. The relationship between BDI score and migraine frequency was present in both aura-present (P trend = .001) and aura-absent subgroups (P trend = .029). Higher migraine frequency, either with or without auras, correlated with higher symptom scores of anxiety and depression.


Evaluation of nationwide referral pathways, investigation and treatment of suspected cauda equina syndrome in the United Kingdom.
Cauda equina syndrome (CES) is a spinal emergency with clinical symptoms and signs that have low diagnostic accuracy. National guidelines in the United Kingdom (UK) state that all patients should undergo an MRI prior to referral to specialist spinal units and surgery should be performed at the earliest opportunity. We aimed to evaluate the current practice of investigating and treating suspected CES in the UK. A retrospective, multicentre observational study of the investigation and management of patients with suspected CES was conducted across the UK, including all patients referred to a spinal unit over 6 months between 1st October 2016 and 31st March 2017. A total of 28 UK spinal units submitted data on 4441 referrals. Over half of referrals were made without any previous imaging (n = 2572, 57.9%). Of all referrals, 695 underwent surgical decompression (15.6%). The majority of referrals were made out-of-hours (n = 2229/3517, 63.4%). Patient location and pre-referral imaging were not associated with time intervals from symptom onset or presentation to decompression. Patients investigated outside of the spinal unit experienced longer time intervals from referral to undergoing the MRI scan. This is the largest known study of the investigation and management of suspected CES. We found that the majority of referrals were made without adequate investigations. Most patients were referred out-of-hours and many were transferred for an MRI without subsequently requiring surgery. Adherence to guidelines would reduce the number of referrals to spinal services by 72% and reduce the number of patient transfers by 79%.


WITHDRAWN: Kinin-enhancing drugs for unexplained subfertility in men.
Oligo-astheno-teratospermia (sperm of low concentration, reduced motility and increased abnormal morphology) of unknown cause is common and the need for treatment is felt by patients and doctors alike. As a result, a variety of empirical, non-specific treatments have been used in an attempt to improve semen characteristics and fertility. One suggested treatment for idiopathic oligo- and/or asthenospermia is the administration of kallikrein (kallidinogenase), a kinin-releasing enzyme (or kininogenase). Although the kinin system may also be involved in the regulation of spermatogenesis in vivo, a clear mechanism of action is missing. Multiple suggestions on how an increase in kinin levels in the genital tract influences spermatogenesis at the testicular levels have been made by various authors. To determine whether treatment of the male with drugs enhancing kinin levles increases pregnancy rates among couples where failure to conceive has been attributed to idiopathic oligo- and/or asthenospermia. Effects on sperm parameters and sex hormones were studied as secondary outcomes. The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SIxteen RCTs on the therapeutic use of androgens (clomiphene citrate or tamoxifen) in subfertile men were identified. Six trials were excluded. All data were extracted by the first author. Unclear issues were resolved by consensus with the senior author (RL). Data was combined for meta-analysis using Peto odds ratios for dichotomous data or weighted mean difference for continuous data. X AUTHORS' CONCLUSIONS: This review does not provide conclusive proof of effectiveness. The benefit on pregnancy rates suggested by observational studies and low quality controlled trials is not confirmed by more rigorous trials. A modest effect on sperm motility is technically possible but was again not confirmed in the better quality trials. The clinical significance of a modest increase in sperm motility is uncertain. Untill effectiveness is proven, kallikrein should only be used in the context of clinical trials.


Differential effects of doses and forms of dietary selenium on immune cell numbers in the skin of ultraviolet-irradiated and unirradiated mice.
The effect of three different doses of dietary L-selenomethionine (SM) and sodium selenite (SS) on skin selenium (Se) content, glutathione peroxidase (GPx) activity, Langerhans cell (LC) and mast cell numbers in ultraviolet radiation-B (UVB)-irradiated and unirradiated C3H/HeN mice was determined. After weaning, groups of mice were given Se-deficient, Se-adequate, or Se-high diets. Six weeks later, some animals in each group were exposed to a single UVB dose (acute), while others were exposed three times weekly for the following 40 weeks (chronic). The skin Se content and GPx activity increased in all the Se-supplemented groups, and the latter was not altered by UVB exposure. Generally, the Se-containing diets caused an increase in LC numbers at 6 weeks and a further rise at 40 weeks, but did not prevent the loss induced by acute or chronic UVB radiation. Skin mast cell numbers were highest in animals fed the Se-deficient diet after 6 and 40 weeks. Acute and chronic UVB radiation decreased the mast cell number and dietary Se did not prevent the reduction. While the present study shows that Se plays an important role in governing the number of LCs and mast cells in the skin, no protective effect against the immunomodulating properties of UVB radiation on these cell types was observed. However, this conclusion may only apply to the experimental conditions chosen, and additional studies at different Se dosages and reduced intensities of chronic UVB exposure are required to confirm the results.


Differentiation of Human Induced Pluripotent Stem Cells from Patients with Severe COPD into Functional Airway Epithelium.
Background: Chronic Obstructive Pulmonary Disease (COPD), a major cause of mortality and disability, is a complex disease with heterogeneous and ill-understood biological mechanisms. Human induced pluripotent stem cells (hiPSCs) are a promising tool to model human disease, including the impact of genetic susceptibility. Methods: We developed a simple and reliable method for reprogramming peripheral blood mononuclear cells into hiPSCs and to differentiate them into air−liquid interface bronchial epithelium within 45 days. Importantly, this method does not involve any cell sorting step. We reprogrammed blood cells from one healthy control and three patients with very severe COPD. Results: The mean cell purity at the definitive endoderm and ventral anterior foregut endoderm (vAFE) stages was >80%, assessed by quantifying C-X-C Motif Chemokine Receptor 4/SRY-Box Transcription Factor 17 (CXCR4/SOX17) and NK2 Homeobox 1 (NKX2.1) expression, respectively. vAFE cells from all four hiPSC lines differentiated into bronchial epithelium in air−liquid interface conditions, with large zones covered by beating ciliated, basal, goblets, club cells and neuroendocrine cells, as found in vivo. The hiPSC-derived airway epithelium (iALI) from patients with very severe COPD and from the healthy control were undistinguishable. Conclusions: iALI bronchial epithelium is ready for better understanding lung disease pathogenesis and accelerating drug discovery.


Efficacy and safety of eslicarbazepine acetate monotherapy in patients converting from carbamazepine.
To evaluate the influence of prior use of carbamazepine (CBZ) and other antiepileptic drugs (AEDs) with a putatively similar mechanism of action (inhibition of voltage-gated sodium channels; VGSCs) on seizure outcomes and tolerability when converting to eslicarbazepine acetate (ESL), using data pooled from 2 controlled conversion-to-ESL monotherapy trials (studies: 093-045, 093-046). Adults with treatment-resistant focal (partial-onset) seizures were randomized 2:1 to ESL 1600 or 1200 mg once daily. The primary efficacy endpoint was study exit (meeting predefined exit criteria related to worsening seizure control) versus an historical control group. Other endpoints included change in seizure frequency, responder rate, and tolerability. Endpoints were analyzed for subgroups of patients who received CBZ (or any VGSC inhibitor [VGSCi]) during baseline versus those who received other AEDs. Of 365 patients in the studies, 332 were evaluable for efficacy. The higher risk of study exit in the subgroups that received CBZ (or any VGSCi) during baseline, versus other AEDs, was not statistically significant (hazard ratios were 1.49 for +CBZ vs -CBZ [P = .10] and 1.27 for +VGSCi vs. -VGSCi [P = .33]). Reductions in seizure frequency and responder rates were lower in patients who converted from CBZ or other VGSCi compared with those who converted from other AEDs. There were no notable differences in overall tolerability between subgroups, but the incidence of some adverse events (eg, dizziness, somnolence, nausea) differed between subgroups and/or between treatment periods. Baseline use of CBZ or other major putative VGSC inhibitors did not appear to significantly increase the risk of study exit due to worsening seizure control, or to increase the frequency of side effects when converting to ESL monotherapy. However, bigger improvements in efficacy may be possible in patients converting to ESL monotherapy from an AED regimen that does not include a VGSC inhibitor.


Modification of sexual behavior of Long-Evans male rats by drugs acting on the 5-HT1A receptor.
Modulation of the sexual behavior of male rats by the anxiolytic buspirone (S-20499) and its analog gepirone were compared to the effects of 8-OH-DPAT (or DPAT, a selective 5-HT1A reference agonist), and BMY-7378 (a selective 5-HT1A partial agonist). Long-Evans rats were used; modulation of copulatory behavior and alteration of penile reflexes were examined. Modulation of copulatory behavior was assessed by three indices: frequency and length of intromission, and latency of ejaculation. DPAT, at doses of 1-8 mg/kg, reduced these three indices in a time dependent manner such that the effects peaked at 45 min and normalized at 90 min. The dose-effect relationship (assessed 45 min after DPAT injection) is bell-shaped with an ED50 approximately 1 mg/kg on the ascending limb of the curve. The effects of buspirone (2 mg/kg) and gepirone (2 mg/kg) on copulatory behavior were indistinguishable from control. BMY-7378 alone and in combination with these other 5-HT1A agonists reduced copulatory behavior, though not statistically significant. Penile reflexes, including number of erections, cups and flips, were inhibited by these agents: DPAT>buspirone>gepirone (inactive at 2 mg/kg). Furthermore, the latency period to erection was at least doubled by DPAT (2 mg/kg). Buspirone and gepirone, however, reduced the latency period to erection. BMY-7378 inhibited penile reflexes when administered alone and even more in combination with DPAT or buspirone. Two butyrophenone analogs, spiperone (a 5-HT1A and dopamine D2 antagonist) and haloperidol (a D2 antagonist), were also tested for their interaction with DPAT. Both of these drugs (at 0.25 mg/kg, 60 min after administration) reduced all indices of penile reflexes and copulation. Furthermore, in combination with DPAT (2 mg/kg, 45 min), the effects were synergistic such that sexual activity came nearly to a standstill. These opposing effects on putatively brain originated copulatory behavior and spinal mediated penile reflexes indicate that the effects of buspirone and DPAT on sexual behavior in the male rat may be possible at different parts of the central nervous system. If a tentative shared target site by DPAT and buspirone is the 5-HT1A receptor, than the same 5-HT receptor sub-type at different locations (brain, raphe nuclei, spinal cord and autonomic ganglia) may modulate rat sexual behavior in opposing ways.


Development and Functionality of a Parsimonious Digital Food Frequency Questionnaire for a Clinical Intervention among an Indigenous Population.
Nutrition-related chronic diseases are a major problem among Indigenous populations. Appropriate dietary intake assessment tools are needed for nutritional surveillance and intervention; however, tools designed to measure the habitual dietary intake of Indigenous persons are largely lacking. We developed a digital food frequency questionnaire (FFQ) to measure habitual consumption among Australian Aboriginal adults and support personalized nutrition counseling. The primary contributors to energy, select nutrients, and inter-person variation (83 food groups) were identified from nationally representative 24 h recall (24HR) data, and they accounted for >80% of the total intake and inter-person variation of the nutrients of interest. Based on community input, a meal-based FFQ format was adopted, with a main food/beverage list of 81 items and the capacity to report on >300 additional items via the digital platform. The nutrient database was based on the Australian Food and Nutrient Database. Data for the first 60 study participants (70% female; median age: 48 years) were used to assess the FFQ's utility. The participants' median [IQR] reported energy intake (10,042 [6968-12,175] kJ/day) was similar to their median [IQR] estimated energy expenditure (10,197 [8636-11,551] kJ/day). Foods/beverages on the main FFQ list accounted for between 66% and 90% of the participants' reported energy and nutrient intakes; the remainder came from participant-selected extra items. The digital FFQ platform provides a potentially valuable resource for monitoring habitual dietary intake among Aboriginal adults and supporting chronic disease prevention and management interventions.


Ipriflavone and Ipriflavone loaded albumin nanoparticles reverse lipopolysaccharide induced neuroinflammation in rats.
Neuroinflammation causes neurodegenerative conditions like Alzheimer's disease (AD). Ipriflavone (IP), therapeutic compound to postmenopausal osteoporosis, has limited estrogenic activity and is accounted as AChE inhibitor. The developing of drug delivery systems to enable drug targeting to specific sites increases the drug therapeutic effect. The aim of the present study was to formulate and evaluate ipriflavone loaded albumin nanoparticles (IP-Np) along with free ipriflavone against lipopolysaccharide (LPS) induced neuroinflammation in rats. Neuroinflammation was induced by intra-peritoneal (i.p) injection of LPS (250 μg/kg rat body weight) then treatments were conducted with (1) ipriflavone at two doses 50 mg/kg and 5 mg/kg, (2) IP-Np (5 mg ipriflavone/kg) or (3) IP-Np coated with polysorbate 80 (IP-Np-T80) (5 mg ipriflavone/kg). The alteration of the inflammatory response in male adult Wistar rats' brain hippocampus was investigated by examining associated indices using biochemical and molecular analyses. A significant upsurge in inflammatory mediators and decline in antioxidant status were observed in LPS-induced rats. In one hand, ipriflavone (50 mg/kg), IP-Np and IP-Np-T80 ameliorated LPS induced brain hippocampal inflammation where they depreciated the level of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and enhanced antioxidant status. In another hand, ipriflavone at dose (5 mg/kg) didn't show the same therapeutic effect. The current study provides evidence for the potential neuroprotective effect of ipriflavone (50 mg/kg) against LPS-induced neuroinflammation in rats through its anti-inflammatory and antioxidant activities. Moreover, nanoparticles significantly attenuated neuroinflammation in concentration lower than the effective therapeutic dose of free drug ten times.


A randomized controlled trial comparing two modern total wrist arthroplasties : improved function with stable implants, but high complication rates in non-rheumatoid wrists at two years.
To analyze the short-term outcome of two types of total wrist arthroplasty (TWA) in terms of wrist function, migration, and periprosthetic bone behaviour. A total of 40 patients suffering from non-rheumatoid wrist arthritis were enrolled in a randomized controlled trial comparing the ReMotion and Motec TWAs. Patient-rated and functional outcomes, radiological changes, blood metal ion levels, migration measured by model-based radiostereometric analysis (RSA), bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA), complications, loosening, and revision rates at two years were compared. Patient-Rated Wrist and Hand Evaluation (PRWHE) scores, abbreviated version of the Disabilities of the Arm, Shoulder and Hand questionnaire scores, and pain improved similarly and significantly in both groups. Wrist motion improved significantly in the Motec group only, and forearm rotation in the ReMotion group only. Cobalt (Co) and chromium (Cr) blood ion levels were significantly higher in the metal-on-metal (MoM) Motec group than in the metal-on-polyethylene (MoP) ReMotion group. Mean total translation was 0.65 mm (95% confidence interval (CI) 0.26 to 1.12) and 0.27 mm (95% CI 0.14 to 0.47) for the ReMotion carpal and radial components, and 0.32 mm (95% CI 0.22 to 0.45) and 0.26 mm (95% CI 0.20 to 0.34) for the Motec metacarpal and radial components, respectively. Apart from dorsal and volar tilts, which were significantly higher for the radial ReMotion than for the Motec component, no significant differences in absolute migration occurred. BMD around the radial components never returned to baseline. Almost one-third of patients required reoperation due to complications. Two ReMotion implants were revised to Motec TWAs due to carpal component loosening, and three Motec MoM articulations were revised to metal-on-polyether ether ketone due to painful synovitis. Both implants provided matched function and were stable at short-term follow-up, but with a high complication rate. This procedure should be restricted to specialist centres undertaking prospective analysis until its role is clarified.Cite this article: Bone Joint J 2022;104-B(10):1132-1141.


Ultrathin, freestanding, stimuli-responsive, porous membranes from polymer hydrogel-brushes.
The fabrication of freestanding, sub-100 nm-thick, pH-responsive hydrogel membranes with controlled nano-morphology, based on modified poly(hydroxyethyl methacrylate) (PHEMA) is presented. Polymer hydrogel-brush films were first synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) and subsequently detached from silicon substrates by UV-induced photo-cleavage of a specially designed linker within the initiator groups. The detachment was also assisted by pH-induced osmotic forces generated within the films in the swollen state. The mechanical properties and morphology of the freestanding films were studied by atomic force microscopy (AFM). Inclusion of nanopores of controlled diameter was accomplished by performing SI-ATRP from initiator-coated surfaces that had previously been patterned with polystyrene nanoparticles. Assembly parameters and particle sizes could be varied, in order to fabricate nanoporous hydrogel-brush membranes with tunable pore coverage and characteristics. Additionally, due to the presence of weak polyacid functions within the hydrogel, the membranes exhibited pH-dependent thickness in water and reversible opening/closing of the pores.


Identification of 5-Methoxy-2-(Diformylmethylidene)-3,3-Dimethylindole as an Anti-Influenza A Virus Agent.
Influenza virus is estimated to cause 3-5 million severe complications and about 250-500 thousand deaths per year. Different kinds of anti-influenza virus drugs have been developed. However, the emergence of drug resistant strains has presented a big challenge for efficient antiviral therapy. Indole derivatives have been shown to exhibit both antiviral and anti-inflammatory activities. In this study, we adopted a cell-based system to screen for potential anti-IAV agents. Four indole derivatives (named 525A, 526A, 527A and 528A) were subjected to the antiviral screening, of which 526A was selected for further investigation. We reported that pre-treating cells with 526A protects cells from IAV infection. Furthermore, 526A inhibits IAV replication by inhibiting the expression of IAV genes. Interestingly, 526A suppresses the activation of IRF3 and STAT1 in host cells and thus represses the production of type I interferon response and cytokines in IAV-infected cells. Importantly, 526A also partially blocks the activation of RIG-I pathway. Taken together, these results suggest that 526A may be a potential anti-influenza A virus agent.


Investigations of the signaling cascade involved in diuretic hormone stimulation of Malpighian tubule fluid secretion in Rhodnius prolixus.
In insects, the excretory system is comprised of the Malpighian tubules (MTs) and the hindgut, which collectively function to maintain ionic and osmotic balance of the haemolymph and rid the organism of toxic compounds or elements in excess. Secretion by the Malpighian tubules of insects is regulated by a variety of hormones including peptidergic factors as well as biogenic amines. In Rhodnius prolixus, two endogenous diuretic hormones have been identified; the biogenic amine serotonin (5-hydroxytryptamine, 5-HT) and the corticotropin releasing factor-related peptide, RhoprCRF. Both factors significantly increase secretion by MTs and are known to elevate intracellular levels of cAMP. Interestingly, applying sub-maximal doses of these two diuretic factors in combination on isolated MTs in vitro reveals synergistic effects as rates of fluid secretion are significantly higher than would be expected if rates of secretion from MTs treated with each factor alone were summed. This observed synergism suggests that different downstream targets may be activated by the two diuretic factors, but that some cellular elicitors may be shared since cAMP is elevated in response to either diuretic hormone. This study investigated the signaling cascade involved in the diuretic hormone regulation of Malpighian tubule fluid secretion. Bioassays were performed in physiological as well as modified salines (e.g. calcium-free) alone or in the presence of a variety of pharmacological compounds that interfere with prospective intracellular targets, such as the apical cation/H(+) exchanger. Intriguingly, only amiloride yielded differential effects on the two diuretics with 5HT-stimulated secretion being blocked, whereas in contrast, RhoprCRF-stimulated secretion was unaffected. In addition, experiments examining the role of extracellular and intracellular calcium on fluid secretion rate showed that both diuretics are dependent on intracellular calcium availability. Finally, fluid secretion stimulated by either diuretic hormone was also sensitive to inhibition of cAMP-dependent protein kinase A. Taken together, these results suggest that each diuretic hormone activates pathways dependent upon intracellular calcium and cAMP.


Forskolin carbamates: binding and activation studies with type I adenylyl cyclase.
Three series of analogs were regioselectively prepared from a protected forskolin precursor to afford 7-carbamoyl-7-desacetylforskolins (series 1), 6-carbamoyl-7-desacetylforskolins (series 2), and 6-carbamoylforskolins (series 3). The analogs were pharmacologically evaluated for binding (IC50) to and activation (EC50) of type I adenylyl cyclase in membranes from stably transfected Sf9 cell lines expressing a single adenylate cyclase subtype. The following ranges were determined for the IC50's and EC50's of each individual series: series 1, IC50 = 43-1600 nM, EC50 = 0.5-9.6 microM; series 2, IC50 = 65-680 nM, EC50 = 0.63-6.5 microM; series 3, IC50 = 21-271 nM, EC50 = 0.5-8.1 microM (forskolin IC50 = 41 nM and EC50 = 0.5 microM). Activation paralleled binding; however, some analogs exhibited poor binding and good activation whereas others demonstrated good binding but poor activation. Steric bulk tended to diminish binding and activation when at the 6- or 7-position, although bulk was accommodated at the 6-position if the 7-site was reacetylated. Acylation of the 7-position by the carbamoyl linker or acetyl was important for obtaining good binding and activation; however, the effect was more pronounced with binding. For both binding and activation, small, linear, lipophilic substituents (propyl, allyl, isopropyl) are well tolerated at the 7-position but less so in the 6-position, even when the 7-site is reacetylated. Planar aromatic moieties (phenyl and 2-pyridinyl) demonstrated moderate to good potency for binding and activation when located at either the 6- or 7-positions. There is an overall trend toward increasing potency for both binding and activation with polar substituents.


Functional crackers: incorporation of the dietary fibers extracted from citrus seeds.
Dietary fibers extracted from defatted press meals of orange and grapefruit seeds were used in the production of crackers and the crackers were evaluated for physicochemical, textural, sensory properties, and bioactive compounds. The unexplored fibers (grapes seeds and orange seeds) and wheat fiber were added at 2.9% levels. The effects of incorporation of different fibers on composition and consumer acceptability were evaluated. The textural stability during storage was also monitored. Although there were no significant differences for proximate compositions; fiber contents, antioxidant capacities, and phenolics compositions were significantly higher in the crackers containing orange seeds fiber. Sensory analysis showed that taste/flavor attribute scores were low, while appearance scores were equal to the control sample. The fracturability, hardness and water activity values remain constant up to 90 days storage at room temperature. The crackers containing orange seed and grapes seed fiber could provide some health benefits to consumers due to their high fiber and flavonoid contents.


Therapeutic hypothermia in brain injuries and related diseases.
Mild or moderate hypothermia is a useful therapeutic method for improving the outcome for patients who have suffered brain injuries including traumatic brain injury and strokes, as well as secondary damages such as neuroinflammation and edema. The feasibility and safety of therapeutic hypothermia have been evaluated in experimental settings and clinical trials. Because the efficacy of therapeutic hypothermia, however, is dependent on the cooling method, application time, duration, and strategies, a continuous effort for obtaining optimal standard protocols of controlling temperature and the documentation of hypothermic devices and drugs are required. Here, we discuss the most recent hypothermia applications in brain injury and related diseases, and review the latest patents in hypothermic devices and cannabinoid-based hypothermia-inducing drugs. Ultimately, we conclude that a localized hypothermia may overcome the limitations, risks, and side effects of systemic hypothermia, and the combination of local hypothermia and drug treatment may improve clinical benefits.


Targeted Radionuclide Therapy: An Evolution Toward Precision Cancer Treatment.
This article reviews recent developments in targeted radionuclide therapy (TRT) approaches directed to malignant liver lesions, bone metastases, neuroendocrine tumors, and castrate-resistant metastatic prostate cancer and discusses challenges and opportunities in this field. TRT has been employed since the first radioiodine thyroid treatment almost 75 years ago. Progress in the understanding of the complex underlying biology of cancer and advances in radiochemistry science, multimodal imaging techniques including the concept of "see and treat" within the framework of theranostics, and universal traction with the notion of precision medicine have all contributed to a resurgence of TRT.


Pentatricopeptide repeat proteins in Trypanosoma brucei function in mitochondrial ribosomes.
The pentatricopeptide repeat (PPR), a degenerate 35-amino-acid motif, defines a novel eukaryotic protein family. Plants have 400 to 500 distinct PPR proteins, whereas other eukaryotes generally have fewer than 5. The few PPR proteins that have been studied have roles in organellar gene expression, probably via direct interaction with RNA. Here we show that the parasitic protozoan Trypanosoma brucei encodes 28 distinct PPR proteins, an extraordinarily high number for a nonplant organism. A comparative analysis shows that seven out of eight selected PPR proteins are mitochondrially localized and essential for oxidative phosphorylation. Six of these are required for the stabilization of mitochondrial rRNAs and, like ribosomes, are associated with the mitochondrial membranes. Furthermore, one of the PPR proteins copurifies with the large subunit rRNA. Finally, ablation of all of the PPR proteins that were tested induces degradation of the other PPR proteins, indicating that they function in concert. Our results show that a significant number of trypanosomal PPR proteins are individually essential for the maintenance and/or biogenesis of mitochondrial rRNAs.


Plasma nitric oxide concentrations and nitric oxide synthase gene polymorphisms in coronary artery disease.
Plasma NOx (nitrate and nitrite) is a stable end product of the vasodilator NO. Several polymorphisms in the endothelial constitutive NO synthase (ecNOS) gene have been reported, including the 4a/4b VNTR polymorphism in intron 4, the E298D mutation in exon 7, and the G10-T polymorphism in intron 23. The aims of this study were to examine plasma NOx in patients with coronary artery disease (CAD) and to assess the association between plasma NOx concentrations and the three ecNOS gene polymorphisms. Plasma NOx was measured in samples from 128 healthy controls and from 110 CAD patients at least 2 months after myocardial infarction. Three genetic polymorphisms that are known or have been suggested to be associated with plasma NOx concentration were also analyzed by PCR-restriction fragment length polymorphism. Median plasma NOx was significantly higher (P <0.001) in CAD patients (95.9 micromol/L) than in controls (73.8 micromol/L). Furthermore, the median plasma NOx was significantly higher (P <0.001) in hypertensive CAD patients (116.0 micromol/L) than in controls and normotensive CAD patients (86.0 micromol/L). The G-allele frequency of the G10-T polymorphism in intron 23 was significantly higher in CAD patients than in controls. Other polymorphisms showed no differences in allelic frequencies among the control and CAD groups. In controls, individuals with the E298D mutation in exon 7 (136.1 micromol/L) showed significantly higher (P = 0.001) median plasma NOx than those without this mutation (64.5 micromol/L). Plasma NOx was higher in hypertensive CAD patients than in normotensive CAD patients and controls. The E298D polymorphism of the ecNOS gene was associated with increased plasma NOx. Further study is needed to understand the gene expression and enzyme activity of ecNOS and their association with genotypes.


Effects of interleukin-1β on human follicular dendritic cell-secreted protein gene expression in periodontal ligament cells.
Follicular dendritic cell-secreted protein (FDC-SP) is expressed in FDCs, human periodontal ligament (HPL) cells, and junctional epithelium. To evaluate the effects of interleukin-1 beta (IL-1β) on FDC-SP gene expression in immortalized HPL cells, FDC-SP mRNA and protein levels in HPL cells following stimulation by IL-1β were measured by real-time polymerase chain reaction and Western blotting. Luciferase (LUC), gel mobility shift, and chromatin immunoprecipitation (ChIP) analyses were performed to study the interaction between transcription factors and promoter regions in the human FDC-SP gene. IL-1β (1 ng/mL) induced the expression of FDC-SP mRNA and protein levels at 3 h, and reached maximum levels at 12 h. IL-1β increased LUC activities of constructs (-116FDCSP - -948FDCSP) including the FDC-SP gene promoter. Transcriptional inductions by IL-1β were partially inhibited by 3-base-pair (3-bp) mutations in the Yin Yang 1 (YY1), GATA, CCAAT-enhancer-binding protein2 (C/EBP2), or C/EBP3 in the -345FDCSP. IL-1β-induced -345FDCSP activities were inhibited by protein kinase A, tyrosine-kinase, mitogen-activated protein kinase (MEK)1/2, and PI3-kinase inhibitors. The results of gel shift and ChIP assays revealed that YY1, GATA, and C/EBP-β interacted with the YY1, GATA, C/EBP2, and C/EBP3 elements that were increased by IL-1β. These studies demonstrate that IL-1β increases FDC-SP gene transcription in HPL cells by targeting YY1, GATA, C/EBP2, and C/EBP3 in the human FDC-SP gene promoter.


Efficient induction of differentiation and growth inhibition in IDH1 mutant glioma cells by the DNMT Inhibitor Decitabine.
Mutation in the IDH1 or IDH2 genes occurs frequently in gliomas and other human malignancies. In intermediate grade gliomas, IDH1 mutation is found in over 70% of tumors. These mutations impart the mutant IDH enzyme with a neomorphic activity - the ability to synthesize 2-hydroxyglutarate (2-HG). This ability leads to a reprogramming of chromatin state, a block in differentiation, and the establishment of the glioma hypermethylator phenotype (G-CIMP). It has been hypothesized but not proven that the extensive DNA methylation that occurs in G-CIMP tumors helps maintain and "lock in" glioma cancer cells in a dedifferentiated state. Here, we tested this hypothesis by treating patient derived IDH1 mutant glioma initiating cells (GIC) with non-cytotoxic, epigenetically targeted doses of the DNMT inhibitor decitabine. Global methylome analysis of treated IDH1 mutant GICs showed that DAC treatment resulted in reversal of DNA methylation marks induced by IDH and the re-expression of genes associated with differentiation. Accordingly, treatment of IDH1 mutant glioma cells resulted in a dramatic loss of stem-like properties and efficient adoption of markers of differentiation, effects not seen in decitabine treated IDH wild-type GICs. Induction of differentiation was much more efficient than that seen following treatment with a specific inhibitor of mutant IDH enzyme (Agios). Decitabine also decreased replicative potential and tumor growth in vivo. Reexpression of polycomb regulated genes accompanied these DAC-induced phenotypes. In total, our data indicates that targeting the pathologic DNA methylation in IDH mutant cells can reverse mutant IDH induced hypermethylation and block in differentiation and promote tumor control. These findings have substantial impact for exploring new treatment strategies for patients with IDH mutant gliomas.


[Severe cardiac disease in scleroderma].
A 36 year-old female with a diagnosis of systemic sclerosis suffered from an acute episode of skeletal myositis and refractory heart failure with cardiogenic shock. Despite immunosuppressive treatment with high doses of corticosteroids and hemodynamic support she died five days after admission. The autopsy showed cardiac myocytolysis, myocyte necrosis and contraction band necrosis. There are very few reported cases of severe heart failure associated to systemic sclerosis.


Functional amino acids in the development of the pig placenta.
The mammalian placenta is essential for supplying nutrients (e.g., amino acids and water) and oxygen from the mother to fetus and for removing fetal metabolites (e.g., ammonia and CO2 ) from fetus to mother. Thus, placental growth and development are determinants of fetal survival, growth, and development. Indeed, low birth weight is closely associated with reduced placental growth. Providing gestating gilts or sows with dietary supplementation of arginine and glutamine, increases placental growth (including vascular growth), improves embryonic/fetal growth and survival, and reduces the large variation in birth weight among litters. These two amino acids serve as building blocks for tissue protein as well as substrates for the production of polyamines and nitric oxide, which stimulate DNA and protein synthesis and angiogenesis and vascular growth in the placenta. These recent findings not only greatly advance the field of mammalian amino acid metabolism and nutrition, but also provide practical, mechanism-based methods to enhance reproductive efficiency in swine. These results may also help improve embryonic/fetal survival and growth in other livestock species (e.g., sheep and cattle) and in humans.


ITGA6 is directly regulated by hypoxia-inducible factors and enriches for cancer stem cell activity and invasion in metastatic breast cancer models.
Hypoxia-inducible factors (HIFs) are well-established mediators of tumor growth, the epithelial to mesenchymal transition (EMT) and metastasis. In several types of solid tumors, including breast cancers, the HIFs play a critical role in maintaining cancer stem cell (CSC) activity. Thus, we hypothesized that HIFs may also regulate transcription of markers of breast CSC activity. One approach to enrich for breast cells with stem-like phenotypes is FACS sorting, in which sub-populations of live cells are gated based on the expression of cell surface antigens, including various integrin subunits. Integrin alpha 6 (ITGA6; CD49f) is routinely used in combination with other integrin subunits to enrich for breast stem cells by FACS. Integrins not only mediate interactions with the extracellular matrix (ECM), but also drive intracellular signaling events that communicate from the tumor microenvironment to inside of the tumor cell to alter phenotypes including migration and invasion. We used two models of metastatic breast cancer (MBC), polyoma middle T (MMTV-PyMT) and MDA-MB-231 cells, to compare the expression of ITGA6 in wild type and knockout (KO) or knockdown cells. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays verified that ITGA6 is a direct HIF transcriptional target. We also used FACS sorting to enrich for CD49f (+) cells to compare tumorsphere formation, tumor initiating cell activity, invasion and HIF activity relative to CD49f(neg or low) cells. Knockdown of ITGA6 significantly reduced invasion, whereas re-expression of ITGA6 in the context of HIF knockdown partially rescued invasion. A search of public databases also revealed that ITGA6 expression is an independent prognostic factor of survival in breast cancer patients. We report that ITGA6 is a HIF-dependent target gene and that high ITGA6 expression enhances invasion and tumor-initiating cell activities in models of MBC. Moreover, cells that express high levels of ITGA6 are enriched for HIF-1α expression and the expression of HIF-dependent target genes. Our data suggest that HIF-dependent regulation of ITGA6 is one mechanism by which sorting for CD49f (+) cells enhances CSC and metastatic phenotypes in breast cancers. Our results are particularly relevant to basal-like breast cancers which express higher levels of the HIFα subunits, core HIF-dependent target genes and ITGA6 relative to other molecular subtypes.


Kidney transplantation in patients with severe preoperative hypertension.
Severe systemic hypertension (HTN) is a risk factor for perioperative cardiovascular complications; however, its impact at the time of kidney transplantation (KTX) is not well defined. A retrospective cohort study of adult kidney-only transplant recipients between October 2009 and December 2012 was performed to examine outcomes between patients with (n = 111) and without (n = 98) severe preoperative HTN defined as SBP > 180 or DBP > 110 mmHg. Recipients with severe HTN were older (56.7 ± 13.0 vs. 53.5 ± 12.4 yr, p = 0.07) and significantly more likely to receive an expanded criteria donor kidney (32.7% vs. 12.2%, p = 0.02). No patients developed hypertensive crisis, intracranial hemorrhage, or life threatening ventricular arrhythmias within 30 d post-transplantation; however, three patients with severe HTN had cardiac events: two with demand ischemia and one with decompensate heart failure. Two patients in the control group had decompensated heart failure. There were no differences between the groups in terms of cardiac event (2.7% vs. 2.0%, p = 0.75), one-yr patient survival (98.2% vs. 98.0%, p = 0.90) or graft survival (90.1% vs. 92.9%, p = 0.48), nadir creatinine > 2 mg/dL (4.6% vs. 6.2%, p = 0.62), length of stay > 6 d (37.8% vs. 35.7%, p = 0.75), and DGF (52.3% vs. 63.3%, p = 0.11). Our results suggest that severe preoperative HTN should not be considered an absolute contraindication to kidney transplant in patients who are otherwise clinically stable.


Seeing is Believing? Preoperative Magnetic Resonance Imaging for Pressure Ulcers: Implications for Surgical Management.
Pressure ulcers represent a particularly difficult disease process and remain a financially important entity. The underlying bone in advanced ulcers may harbor osteomyelitis. Radiologic diagnosis of osteomyelitis is confounded by chronic pressure and shear. We sought to determine the test characteristics of preoperative magnetic resonance imaging (MRI) in the diagnosis of osteomyelitis compared to intraoperative bone culture. A retrospective review of patients undergoing flap reconstruction who had preoperative MRI and intraoperative bone cultures between 1995 and 2015 was included. Recorded variables included age, sex, level of spinal cord injury and duration, preoperative MRI interpretation, microbiologic bone culture, smoking history, comorbidities, colostomy or urostomy, healing time, complications, length of stay, and discharge facility. A total of 152 patients (175 flaps) were reconstructed, of which 41 patients (73 flaps) met inclusion criteria. Most patients were male (82.2%) with an average age of 50.4 years. Overall complication rate was 32.4% (n = 23) of which 34.7% (n = 8) were major. Positive and negative predictive MRI values were 84.6% and 16.7%, respectively. There were no significant differences in healing time or complication rate in those with or without osteomyelitis. Intraoperative growth was associated with decreased postoperative complications (hazard ratio = 0.361; P = 0.037). Test properties of MRI for diagnosis of osteomyelitis in patients with chronic pressure ulcers have limited ability to diagnose osteomyelitis and do not aid in surgical management, but do increase health-care expense. The diagnosis of osteomyelitis by intraoperative bone cultures does not predict inferior outcomes and paradoxically may be associated with fewer postoperative complications.


Husbands' Awareness of Sanhujori, Needs for Education and Family Strength.
We conducted a descriptive study to: 1) understand how husbands are aware of the importance of Sanhujoiri(i.e, Korean traditional postpartum care) and 2) identify their needs to learn how to take care of postpartum women and new-borns and to strengthen family bonding. The subjects consisted of 123 husbands who had children under the age of 5 years. Data were analyzed with t-test, one-way ANOVA, Scheffe's test, and Pearson's correlation. The mean age of subjects was 37.1+/-4.63 years. The mean scores of awareness, needs for education on postpartum mother and newborn care, and family strength were 4.57+/-1.18, 4.61+/-1.18, 4.92+/-1.07, and 4.01+/-0.95, respectively, indicating higher scores. There were statistically significant differences in awareness (F=5.08, p<.05), newborn care (F=3.70, p<.05), and family strength (F=4.64, p<.05) by husband's role in Sanhujori There was a positive correlation among study variables. This study shows that even though husbands want to participate in Sanhujori, they do not have enough information on Sanhujori and mother/newborn care and adequate paternity leave. Realistic paternity leave system and effective Couple Centered Childbearing (from pregnancy to postpartum) Education Program CCCEP development are required to help husbands' participation in mother/newborn care with confidence and competence in home based Sanhujori.


Efficacy analysis and hemodynamic changes of hematological system diseases after PICC chemotherapy.
The aim of the study was to investigate the efficacy analysis and hemodynamic changes of hematological system diseases after peripherally inserted central catheter (PICC) chemotherapy. From March 2017 to March 2020, patients with hematological diseases who received chemotherapy in our hospital were selected. The experimental group consisted of 80 patients with PICC, and the control group consisted of 70 patients with routine intravenous injection. The indwelling time, prevalence rate of adverse reactions, hemodynamic indicators before and after chemotherapy and compliance of patients in the two groups were compared. The longest time and average time of indwelling catheter in experimental group were significantly higher than those in control group (p<0.05). The total prevalence rate of adverse reactions in experimental group (3.75%) was significantly lower than that in control group (20%) (p<0.05). The compliance of patients with PICC in experimental group was significantly better than that of patients with superficial intravenous injection in control group. There was no significant difference in hemodynamic indicators between experimental group and control group before and after chemotherapy (p>0.05). There was significant difference in high-shear whole blood indicators, low-shear whole blood viscosity and high-shear whole blood viscosity after chemotherapy (p<0.05), while there was no significant difference in changes of other hemodynamic indicators (p>0.05) or in hemorheological indicators between the two groups after chemotherapy (p>0.05). The total prevalence of adverse reactions in hematological diseases by PICC infusion is lower than that by superficial vein infusion, and the catheter has no significant influence on hemodynamic indicators in patients. Therefore, PICC catheter is worthy of application and promotion in hematological diseases.


Simulations for inverse radiation therapy treatment planning using a dynamic MLC algorithm.
The inverse radiation treatment planning model for a dynamic multileaf collimator (MLC) is used to find the optimal solution of planning problem. The model for dynamic MLC is explained in Tervo et al (2003 Appl. Math. Comput. 135 227-50). The advantage of this model is that it optimizes leaf velocity parameters directly. Our algorithm uses a gradient-based local optimization method. Two patient cases, prostate carcinoma and tonsilla carcinoma, are studied. Field arrangements are pre-selected and velocity parameters for MLC leaves are optimized to obtain the prescribed dose in the patient space. In both simulated cases, high dose distribution conforms the planning target volume well and organs-at-risk are saved in most parts. Simulations show that the model has its functionality in patient treatments, although it is still formal and needs further development.


Human alpha 1,3/4 fucosyltransferases. Characterization of highly conserved cysteine residues and N-linked glycosylation sites.
Human alpha1,3 fucosyltransferases (FucTs) contain four highly conserved cysteine (Cys) residues, in addition to a free Cys residue that lies near the binding site for GDP-fucose (Holmes, E. H., Xu, Z. , Sherwood, A. L., and Macher, B. A. (1995) J. Biol. Chem. 270, 8145-8151). The participation of the highly conserved Cys residues in disulfide bonds and their functional significance were characterized by mass spectrometry (MS) analyses and site-directed mutagenesis, respectively. Among the human FucTs is a subset of enzymes (FucT III, V, and VI) having highly homologous sequences, especially in the catalytic domain, and Cys residues in FucT III and V were characterized. The amino acid sequence of FucT III was characterized. Peptides containing the four conserved Cys residues were detected after reduction and alkylation, and found to be involved in disulfide bonds. The disulfide bond pattern was characterized by multiple stage MS analysis and the use of Glu-C protease and MS/MS analysis. Disulfide bonds in FucT III occur between Cys residues (Cys(81) to Cys(338) and Cys(91) to Cys(341)) at the N and C termini of the catalytic domain, bringing these ends close together in space. Mutagenesis of highly conserved Cys residues to Ser in FucT V resulted in proteins lacking enzymatic activity. Three of the four mutants have molecular weights similar to wild type enzyme and maintained an ability to bind GDP, whereas the other (Cys(104)) produced a series of lower molecular weight bands when characterized by Western blot analysis, and did not bind GDP. FucTs have highly conserved, potential N-linked sites, and our mass spectrometry analyses demonstrated that both N-linked sites are modified with oligosaccharides.


Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins.
Green fluorescent protein (GFP) is widely used as a reporter gene in both prokaryotes and eukaryotes. However, the fluorescence levels of wild-type GFP (wtGFP) are not bright enough for fluorescence-activated cell sorting or flow cytometry. Several GFP variants were generated that are brighter or have altered excitation spectra when expressed in prokaryotic cells. We engineered two GFP genes with different combinations of these mutations, GFP(S65T,V163A) termed GFP-Bex1, and GFP(S202F,T203I,V163A) termed GFP-Vex1. Both show enhanced brightness and improved signal-to-noise ratios when expressed in mammalian cells and appropriately excited, compared with wtGFP. Each mutant retains only one of the two excitation peaks of the wild-type protein. GFP-Bex1 excites at 488 nm (blue) and GFP-Vex1 excites at 406 nm (violet), both of which are available laser lines. Excitation at these wavelengths allows for the independent analyses of these mutants by fluorescence-activated cell sorting, permitting simultaneous, quantitative detection of expression from two different genes within single mammalian cells.


[Long-term effects of structum administration (according to data from multicenter trial)].
To evaluate duration of a clinical response to the drug structum in patients with osteoarthrosis (OA) of the knee and hip joints as well as structum effects on OA course and that of concomitant diseases. The duration of a clinical response to structum after the end of the treatment and its effect on OA and concominant diseases course were studied for 12 months in 9 centers participating in the study of the drug efficacy and tolerance in patients with gon- and coxarthrosis in an open multicenter randomized controlled 6-month trial. Out of 555 patients with OA of the knee and hip joints enrolled in the first study, the examination covered 373 patients: 159 patients of the test group treated for 6 months with structum and 214 controls. By basic clinical parameters the groups were similar. Clinical examination was made after structum treatment and 12 months later and included assessment of the number of exacerbations, hospitalizations, outpatient consultations, days of temporary disability for OA, pain in the joints while walking and at rest by the visual scale, Leken's functional index, x-ray pictures of the joints, administration of nonsteroidal anti-inflammatory drugs (NSAID), exacerbations of concomitant diseases (gastrointestinal diseases, arterial hypertension, ischemic heart disease). An overall functional Leken's index in patients with gon- and coxarthrosis given structum 12 months after the treatment did not reach the initial values as well as pain and daily need in NSAID. Structum effect in patients with knee joint OA persisted for 4.6 months, in hip joint OA--4.1 months; in patients with stage I-II the effect lasted longer than in stage III (5.2 and 4.6 months vs 4.17 and 3.24 months, respectively. Even short-term therapy with structum reduced the number of further exacerbations, hospitalizations and visits to their doctor. 12 months after structum therapy the effect persisted in 40% patients. Frequency of exacerbations of the concomitant diseases was less than in patients on continuous NSAID. Structum is a highly effective drug against OA as it acts long, reduces frequency of exacerbations, hospitalizations, visits to the doctor, duration of disability, NSAID requirement and improves the course of some concomitant diseases.


Deaths from falls in children: how far is fatal?
The outcome of injury was determined in 317 children who were brought to a children's trauma center with a history from the caretaker that the child had fallen. Cases in which the clinicians' judgment was that an incorrect history had been given were included along with cases in which the history was not questioned. Seven deaths occurred in 100 children who fell 4 feet or less. One death occurred in 117 children who fell 10 feet to 45 feet. The 7 children who died in short falls all had other factors in their cases which suggested false histories. When children incur fatal injuries in falls of less than 4 feet, the history is incorrect. Long falls with an outdoor component are likely to be reliable data points for studies of children's injuribility.


Pulmonary immunization of chickens using non-adjuvanted spray-freeze dried whole inactivated virus vaccine completely protects against highly pathogenic H5N1 avian influenza virus.
Highly pathogenic avian influenza (HPAI) H5N1 virus is a major threat to public health as well as to the global poultry industry. Most fatal human infections are caused by contact with infected poultry. Therefore, preventing the virus from entering the poultry population is a priority. This is, however, problematic in emergency situations, e.g. during outbreaks in poultry, as there are currently no mass application methods to effectively vaccinate large numbers of birds within a short period of time. To evaluate the suitability of needle-free pulmonary immunization for mass vaccination of poultry against HPAI H5N1, we performed a proof-of-concept study in which we investigated whether non-adjuvanted spray-freeze-dried (SFD) whole inactivated virus (WIV) can be used as a dry powder aerosol vaccine to immunize chickens. Our results show that chickens that received SFD-WIV vaccine as aerosolized powder directly at the syrinx (the site of the tracheal bifurcation), mounted a protective antibody response after two vaccinations and survived a lethal challenge with HPAI H5N1. Furthermore, both the number of animals that shed challenge virus, as well as the level of virus shedding, were significantly reduced. Based on antibody levels and reduction of virus shedding, pulmonary vaccination with non-adjuvanted vaccine was at least as efficient as intratracheal vaccination using live virus. Animals that received aerosolized SFD-WIV vaccine by temporary passive inhalation showed partial protection (22% survival) and a delay in time-to-death, thereby demonstrating the feasibility of the method, but indicating that the efficiency of vaccination by passive inhalation needs further improvement. Altogether our results provide a proof-of-concept that pulmonary vaccination using an SFD-WIV powder vaccine is able to protect chickens from lethal HPAI challenge. If the efficacy of pulmonary vaccination by passive inhalation can be improved, this method might be suitable for mass application.


Providing spiritual care to patients with cancer.
Patients with cancer, oncology nurses, and chaplains agree that nurses should provide spiritual care, but organizational and professional barriers often exist. To overcome these barriers, many nurses need only to complement their professional skill sets of problem solving, critical thinking, caring, and communication with knowledge of some spiritual-care specifics. This article provides basic definitions of religion and spirituality and their relationship as well as suggested religiospiritual assessment strategies, standard spiritual diagnoses, a pragmatic intervention approach, and documentation recommendations.


β-adrenergic stimulation activates early afterdepolarizations transiently via kinetic mismatch of PKA targets.
Sympathetic stimulation regulates cardiac excitation-contraction coupling in hearts but can also trigger ventricular arrhythmias caused by early afterdepolarizations (EADs) in pathological conditions. Isoproterenol (ISO) stimulation can transiently cause EADs which could result from differential kinetics of L-type Ca current (ICaL) vs. delayed rectifier potassium current (IKs) effects, but multiple PKA targets complicate mechanistic analysis. Utilizing a biophysically detailed model integrating Ca and β-adrenergic signaling, we investigate how different phosphorylation kinetics and targets influence β-adrenergic-induced transient EADs. We found that: 1) The faster time course of ICaL vs. IKs increases recapitulates experimentally observed ISO-induced transient EADs (which are due to ICaL reactivation). These EADs disappear at steady state ISO and do not occur during more gradual ISO application. 2) This ICaL vs. IKs kinetic mismatch with ISO can also induce transient EADs due to spontaneous sarcoplasmic reticulum (SR) Ca release and Na/Ca exchange current. The increased ICaL, SR Ca uptake and action potential duration (APD) raise SR Ca to cause spontaneous SR Ca release, but eventual IKs activation and APD shortening abolish these EADs. 3) Phospholemman (PLM) phosphorylation decreases both types of EADs by increasing outward Na/K-ATPase current (INaK) for ICaL-mediated EADs, and reducing intracellular Na and Ca loading for SR Ca-release-mediated EADs. Slowing PLM phosphorylation kinetics abolishes this protective effect. 4) Blocking phospholamban (PLB) phosphorylation has little effect on ICaL-mediated transient EADs, but abolishes SR Ca-release-mediated transient EADs by limiting SR Ca loading. 5) RyR phosphorylation has little effect on either transient EAD type. Our study emphasizes the importance of understanding non-steady state kinetics of several systems in mediating β-adrenergic-induced EADs and arrhythmias.


Immunization with bacterial antigens: edwardsiellosis.
Enteric septicaemia of catfish (ESC), caused by Edwardsiella ictaluri, is the most serious disease affecting commercial catfish culture in the United States. ESC is generally an acute septicaemia that develops very quickly, especially in the temperature range of 22-28 degrees C, with a more chronic disease presentation outside this range. The ability of E. ictaluri to avoid the host's immune system and proliferate into a systemic infection is impressive. Catfish kidney tissue cultured positive for E. ictaluri as soon as 15 minutes following gastric lavage and signs of disease are observed microscopically within two days of immersion challenge, with reported mortalities as early as five days following immersion challenge. Analysis of E. ictaluri antigens by several investigators using SDS-PAGE and colorimetric western blotting with immune catfish has identified as many as 15 immunogenic bands. Analysis using two-dimensional SDS-PAGE and chemiluminescent western blotting identified 14 bands and 25 spots as consistently immunogenic. The strongest immunodominant antigens were reported as 34-37 KD and 60 KD, depending on the study. Lipopolysaccharide is the only purified component of E. ictaluri tested for fish vaccination, and results indicated that very poor protection was induced unless Freund's Complete Adjuvant was used. Because E. ictaluri strains are serologically homogeneous, most studies on vaccination have emphasized killed whole cell preparations and have delivered equivocal results. Although antibodies are produced to a variety of preparations, a positive antibody response does not correlate with protection unless very high titres are achieved. Efficacy of killed products has been demonstrated in field trials, and an orally delivered product has been licensed. However, protection probably relies on booster exposure of the host to E. ictaluri during non-permissive temperatures. As a facultative intracellular pathogen, further studies on vaccination of catfish against E. ictaluri should target products and delivery methods that favour induction of cell mediated immunity.


Properties and applications of antimicrobial peptides in biodefense against biological warfare threat agents.
Recent advances in knowledge of the properties of antimicrobial peptides (AMPs) are reviewed. AMPs are typically small, positively charged, amphipathic peptides that interact electrostatically and non-stereospecifically with the bacterial cell membrane, resulting in its permeabilization and cell death. Classes of AMPs, their mechanisms of action, hemolytic activity, and cytotoxicity towards host cells are discussed. A particular focus is AMPs with potential for use in defense against biological warfare agents. Some AMPs cytotoxic to Bacillus anthracis have been described. Synthesis of these peptides in multivalent form leads to a synergistic increase in antibacterial activity. Strategies to enhance the potency, stability, and selectivity of AMPs are discussed.


Cisplatin plus 5-FU for the treatment of adenocarcinoma of the colon.
Cisplatin plus 5-FU appears to have significant additive activity in various tumors, such as head and neck carcinoma and esophageal cancer. A partial explanation for this may be drug synergism, which has been noted in the L1210 leukemia model. Based on these data, a prospective trial of weekly bolus 5-FU (15 mg/kg) and cisplatin (60 mg/m2) given every 3 weeks was initiated at Indiana University. Forty-one patients, of whom 38 are fully evaluable for response, were treated with these two drugs. Ten partial and one complete response (complete + partial response rate = 29%) were observed in the 38 evaluable patients. Thirteen additional patients had stable disease for greater than or equal to 3 months. The median durations of remission and survival time were 6 and 10.3 months, respectively. Myelosuppression was unusually severe, with granulocyte counts less than 1000/mm3 in 65% of patients, including four patients with granulocyte count nadirs less than 100/mm3. Three patients developed granulocytopenic fever, with two drug-related deaths (sepsis, hyperosmolar coma). Nearly all patients had nausea and vomiting, but this was not a treatment-limiting toxic effect in any patient. Although this combination suggests a higher response rate than usually seen with bolus iv 5-FU in colon cancer, a trial comparing 5-FU alone or with cisplatin to determine whether true synergy exists is currently underway.


MicroRNA miR-590-5p inhibits breast cancer cell stemness and metastasis by targeting SOX2.
SOX2 (Sry-related high-mobility box SOX-2) is a transcription factor, which is essential for maintaining the cancer cell stemness. However, the role of microRNAs targeting SOX2 in cancer cell stemness remains unclear. We examined the effect of miR-590-5p, which targeted SOX2, on the breast cancer cell stemness and metastasis. We predicted and screened microRNA targeting SOX2, and further investigated the regulatory role of miR-590-5p on the level of SOX2 with Western blot, luciferase reporting assay and qRT-PCR analysis. Flow cytometry was performed to detect the effect of miR-590-5p on the breast cancer stem cell population with ALDEFLUOR Assay. We inoculated the breast cancer cells transfected with or without miR-590-5p to NOD/SCID mice to detect the tumorigenicity in vivo. Finally, forty-nine pairs of breast cancer samples and adjacent noncancerous tissues were obtained, and immunohistochemistry (IHC) with SOX2 antibody and qRT-PCR assay were used to quantify the expression of miR-590-5p in breast cancer samples. miR-590-5p significantly downregulated the SOX2 protein expression, and inhibition of miR-590-5p increased SOX2 expression. The luciferase reporter assay indicated that miR-590-5p decreased the SOX2 3'UTR (3' untranslated region) reporter activity but not the luciferase activity of the mutant reporter, in which the binding sites for miR-590-5p were mutated. ALDEFLUOR Assay showed that miR-590-5p significantly decreased breast cancer stem cells population. NOD/SCID nude mice experiments indicated that miR-590-5p significantly inhibited tumorigenicity of breast cancer cells. IHC assay and qRT-PCR suggested that miR-590-5p expression was downregulated in breast cancer patients, and negatively correlated with SOX2. miR-590-5p inhibited breast cancer cell stemness through targeting SOX2. Our study indicated that miR-590-5p might be a useful strategy for breast cancer treatment.


Isolation and whole genome analysis of endospore-forming bacteria from heroin.
Infections caused by endospore-forming bacteria have been associated with severe illness and death among persons who inject drugs. Analysis of the bacteria residing in heroin has thus been biased towards species that affect human health. Similarly, exploration of the bacterial diversity of seized street market heroin correlated with the skin microflora of recreational heroin users insofar as different Staphylococus spp. or typical environmental endospore formers including Bacillus cereus and other Bacilli outside the B. cereus sensu lato group as well as diverse Clostridia were identified. In this work 82 samples of non-street market ("wholesale") heroin originating from the German Federal Criminal Police Office's heroin analysis program seized during the period between 2009 and 2014 were analyzed for contaminating bacteria. Without contact with the end user and with only little contaminations introduced by final processing, adulteration and cutting this heroin likely harbors original microbiota from the drug's original source or trafficking route. We found this drug to be only sparsely populated with retrievable heterotrophic, aerobic bacteria. In total, 68 isolates were retrieved from 49 out of 82 samples analyzed (60% culture positive). All isolates were endospore-forming, Gram-positive Bacilli. Completely absent were non-endospore-formers or Gram-negatives. The three most predominant species were Bacillus clausii, Bacillus (para)licheniformis, and Terribacillus saccharophilus. Whole genome sequencing of these 68 isolates was performed using Illumina technology. Sequence data sets were assembled and annotated using an automated bioinformatics pipeline. Average nucleotide identity (ANI) values were calculated for all draft genomes and all close to identical genomes (ANI>99.5%) were compared to the forensic data of the seized drug, showing positive correlations that strongly warrant further research on this subject.