Assessment of ROS Production in the Mitochondria of Live Cells.
Production of reactive oxygen species (ROS) in the mitochondria plays multiple roles in physiology, and excessive production of ROS leads to the development of various pathologies. ROS in the mitochondria are generated by various enzymes, mainly in the electron transporvt chain, and it is important to identify not only the trigger but also the source of free radical production. It is important to measure mitochondrial ROS in live, intact cells, because activation of ROS production could be initiated by changes in extramitochondrial processes which could be overseen when using isolated mitochondria. Here we describe the approaches, which allow to measure production of ROS in the matrix of mitochondria in live cells. We also demonstrate how to measure kinetic changes in lipid peroxidation in mitochondria of live cells. These methods could be used for understanding the mechanisms of pathology in a variety of disease models and also for testing neuro- or cardioprotective chemicals.


Associations between subgingival plaque bacterial morphotypes and clinical indices?
The aim of the present study was to evaluate possible associations between subgingival plaque bacterial morphotypes, as assessed by darkground microscopy, and clinical indices of routine adult chronic periodontitis. Clinical indices were plaque index (PlI), gingival index (GI), sulcus bleeding index (SBI), papilla bleeding index (PBI), attachment loss (AL), pocket depth (PD) and probeable pocket depth (PPD). Apical border plaque was sampled in vivo and after extraction to test whether direct or indirect sampling affected any such associations. Similarly, pocket depth and attachment loss were also assessed directly and indirectly on the same teeth, in vivo or after extraction. The influence of the type of index used to record inflammation (GI, SBI, PBI) was also assessed, as were the effects of the numbers of sampled subjects and the method of analysis, which comprised the use of transformed and untransformed data and of parametric and non-parametric tests. Data were collected in relation to the approximal surfaces of 44 teeth extracted from 22 adults (2 teeth each) and from 1 pair of contralateral upper anterior or premolar teeth in each of 100 adults, all which untreated routine chronic periodontitis. Selected subjects had greater than or equal to 4 mm probeable pocket depth and/or attachment loss, and radiographic evidence of bone loss in relation to 1 approximal surface on each of 1 pair of contralateral anterior or premolar teeth, or to 2 teeth scheduled for extraction. Plaque preparation and darkground microscopy were as described previously. Insignificant associations (p greater than 0.05) were demonstrated between supragingival plaque (PlI) and periodontal inflammation (GI, SBI, PBI) or destruction (PPD and AL), as well as between inflammation and attachment level. In contrast, significant moderate associations (r = 0.5-0.77) were demonstrated between each of the 3 morphotype groups; spirochaetes, other motiles and cocci. Spirochaetes showed a significant moderate (r = 0.5) positive association with pocket depth with a 2.43% mean increase of spirochaetes for each 1 mm increase of PPD. Although highly significant associations (r = 0.9) were demonstrable between the 3 inflammation indices (GI, SBI, PBI) themselves, only PBI showed significant positive associations (r = 0.3) with spirochaetes and other motiles. Also, PlI showed significant associations with each of the 4 morphotypes (r = 0.3-0.5). The heterogeneity of spirochaetes and other motiles as well as the multiplicity of possible aetiological microbial agents in plaque may have resulted in underestimated associations between subjects as well as undetectable association within a given mouth using only 4 morphotype groups.(ABSTRACT TRUNCATED AT 400 WORDS)


A Bioengineered Human Skin Tissue for the Treatment of Infected Wounds.
Complex skin defects resulting from acute skin trauma and chronic, nonhealing wounds are life-threatening injuries. Infection is one of the most common obstacles to the healing of these types of wounds. Host defense peptides (HDPs) possessing a broad spectrum of activity against microorganisms and serving as innate immune modulators have emerged as potential treatment strategies for infected wounds. The increase in multidrug-resistant clinical bacterial isolates highlights the need for new and innovative anti-infective therapies for the treatment of both acute and chronic skin wounds. To address the critical need for new therapeutic options to reduce infection and improve wound healing, a bioengineered skin substitute (BSS) tissue has been created to act as an anti-infective living human skin tissue that provides enhanced expression of the endogenous HDP, cathelicidin. To generate a BSS exhibiting these antimicrobial properties, the clinically tested NIKS progenitor cells were employed to provide a source of genetically uniform, nontumorigenic, pathogen-free human keratinocytes that are amenable to genetic engineering using nonviral means. Pathogenic bacterial strains are increasingly developing antibiotic resistance, thereby forcing the clinician to use potent antibiotics with deleterious effects on keratinocyte viability and migration. Therefore, an urgent need exists for new wound therapies that can circumvent many of the problems associated with current antibiotic treatments. Enhanced expression of cathelicidin in a genetically engineered human BSS has been shown to inhibit the bacterial growth of a multidrug-resistant clinical strain of Acinetobacter baumannii in vivo, creating a new and innovative therapeutic option for combating these debilitating wound infections while also promoting healing.


T-Cell Subsets as Potential Biomarkers for Hepatobiliary Cancers and Selection of Immunotherapy Regimens as a Treatment Strategy.
Patients with advanced hepatocellular or biliary cancers have a dismal prognosis with limited efficacy from standard systemic therapies. The benefit of precision medicine has so far been limited to a subset of biliary cancers, including FGFR rearrangements; hotspot mutations in IDH1/2, BRAF, and BRCA1/2; and other rare alterations. In contrast, hepatocellular carcinoma, an inflammation-driven cancer with an immune-infiltrated microenvironment, provides a promising opportunity for immunotherapy, compared with the highly desmoplastic immune desert or excluded stromal microenvironment in biliary cancers. The immune contexture in hepatobiliary cancers is mostly immunosuppressive, protumorigenic, and exhausted, which together with low tumor mutation burden and decreased neoantigens provides challenges for immunotherapy. A better understanding of the spatiotemporal profile of T cells within the tumor microenvironment and the dynamic interplay of immune modulators in the context of standard or experimental therapies is crucial to define additional markers of response and design evidence-based combinatorial regimens. This review considers recent literature in this area and highlights promising leads and emerging trends.


Feasibility of alcohol screening among patients receiving opioid treatment in primary care.
Identifying and treating problem alcohol use among people who also use illicit drugs is a challenge. Primary care is well placed to address this challenge but there are several barriers which may prevent this occurring. The objective of this study was to determine if a complex intervention designed to support screening and brief intervention for problem alcohol use among people receiving opioid agonist treatment is feasible and acceptable to healthcare providers and their patients in a primary care setting. A randomised, controlled, pre-and-post design measured feasibility and acceptability of alcohol screening based on recruitment and retention rates among patients and practices. Efficacy was measured by screening and brief intervention rates and the proportion of patients with problem alcohol use. Of 149 practices that were invited, 19 (12.8 %) agreed to participate. At follow up, 13 (81.3 %) practices with 81 (62.8 %) patients were retained. Alcohol screening rates in the intervention group were higher at follow up than in the control group (53 % versus 26 %) as were brief intervention rates (47 % versus 19 %). Four (18 %) people reduced their problem drinking (measured by AUDIT-C), compared to two (7 %) in the control group. Alcohol screening among people receiving opioid agonist treatment in primary care seems feasible. A definitive trial is needed. Such a trial would require over sampling and greater support for participating practices to allow for challenges in recruitment of patients and practices.


[The connections between smoking, mucociliary clearance and airway obstruction].
The connection between smoking habits (pack years, smoker and ex-smoker period), mucociliary clearance (mC) and the obstruction of the respiratory passages was investigated in 17 ex-smokers (7 without bronchitis; 10 with bronchitis) and 48 smokers (13 without bronchitis; 35 with bronchitis). The ex-smokers without chronic bronchitis showed the same clearing behaviour as the healthy non-smokers, but the non-smokers did not have the least obstruction of the respiratory system. The smokers without chronic bronchitis showed a normal lung function, but a worse clearing behaviour than the healthy non- and ex-smokers (p less than 0,01). Compared to volunteers without bronchitic symptoms the ex-smokers and smokers with chronic bronchitis had a bad clearing behaviour (p less than 0,001) and a distinctly obstructive ventilation disorder. Furthermore, it was noticeable that the ex-smokers with persistent bronchitis showed the worst respiratory system obstructions, whilst the smokers with chronic bronchitis had the greatest restriction of the central mC (c-mC) with less respiratory system obstruction than the ex-smokers with chronic bronchitis. The smoking habits (pack years) correlated with the decrease of the mC (p less than 0,01) and the grade of the respiratory system obstruction (p less than 0,001).


Hypothalamus-Pituitary-Adrenal Dysfunction in Cholestatic Liver Disease.
The Hypothalamic-Pituitary-Adrenal (HPA) axis has an important role in maintaining the physiological homeostasis in relation to external and internal stimuli. The HPA axis dysfunctions were extensively studied in neuroendocrine disorders such as depression and chronic fatigue syndrome but less so in hepatic cholestasis, cirrhosis or other liver diseases. The HPA axis controls many functions of the liver through neuroendocrine forward signaling pathways as well as negative feedback mechanisms, in health and disease. This review describes cell and molecular mechanisms of liver and HPA axis physiology and pathology. Evidence is presented from clinical and experimental model studies, demonstrating that dysfunctions of HPA axis are correlated with liver cholestatic disorders. The functional interactions of HPA axis with the liver and immune system in cases of bacterial and viral infections are also discussed. Proinflammatory cytokines stimulate glucocorticoid (GC) release by adrenals but they also inhibit bile acid (BA) efflux from liver. Chronic hepatic inflammation leads to cholestasis and impaired GC metabolism in the liver, so that HPA axis becomes depressed. Recently discovered interactions of GC with self-oscillating transcription factors that generate circadian rhythms of gene expression in brain and liver, in the context of GC replacement therapies, are also outlined.


Continuous production of monoclonal antibody in a packed-bed bioreactor.
In the present study the growth and MAb (monoclonal antibody) production of a mouse x mouse hybridoma cell producing anti-digoxin MAb was evaluated. The hybridoma cells entrapped within the support matrix Fibra-Cel were cultured in batch and continuous mode following special protocols. Cell-culture studies were performed in a 1-litre spinner basket containing 3 g.litre-1 support matrix. Batch culture was operated with the cell density of 42x10(6) cells. During the 7 days of culture, the medium was sampled daily in order to assess glucose and MAb concentrations and the lactate dehydrogenase released into the culture medium. After a culture period of 72 h, the cell density and MAb concentration were found to be 10.4x10(7) cells/3 g of NWPF (non-woven polyester fibre) discs and 250 microg/ml respectively. This yield gradually decreased to 0.55x10(6) cells/3 g of packaging material and 60 microg/ml respectively at the end of the batch culture. In the continuous-culture studies, the batch culture was initially operated for 64.5 h and then continuous flow was started at the dilution rates of 0.15, 0.2, 0.25 and 0.3 day-1 and finally stabilized at 0.25 day-1 within 288 h (12 days). The MAb concentration at steady state was found to be 116-120 microg/day per ml, and the yield of operation was 62.5 mg/day per ml, which was 3.5 times higher than that of batch culture. In conclusion, a packed-bed bioreactor with the support matrix Fibra-Cel, operated in continuous-feeding mode, is more efficient for large-scale MAb production than a batch culture. On the other hand, by using a continuous-culture system, a better supply of nutrients and removal of inhibitory metabolites and proteolytic enzymes was obtained.


Measures of Care Coordination at Inpatient Psychiatric Facilities and the Medicare 30-Day All-Cause Readmission Rate.
Since late 2012, the Medicare Inpatient Psychiatric Facility Quality Reporting (IPFQR) program of the Centers for Medicare and Medicaid Services (CMS) has required inpatient psychiatric facilities to collect and publicly report a suite of quality measures. This study explored the association between facility-level 30-day risk-adjusted all-cause readmission (medical or psychiatric) after psychiatric hospitalization (READM-30-IPF) and care coordination process measures in the IPFQR program. The study used publicly reported IPFQR facility-level performance data of the Hospital Compare Web site for 1,343 inpatient psychiatric facilities, reflecting performance from July 2015 to June 2017. The authors used a cross-sectional design and linear regression models controlling for hospital and community characteristics and using state as fixed effect. The mean±SD facility-level READM-30-IPF was 20%±3%, with substantial variation by facility type, ownership status, rurality, and percentage of racial-ethnic minority residents in the county. Regression results showed that facilities with performance in the top tercile on the measure of 7-day mental health follow-up after discharge had readmission rates significantly lower than facilities in the bottom tercile (coefficient=-0.58, p<0.01), although the magnitude of this difference was small. READM-30-IPF, however, did not vary by facilities' performance on measures of discharge plan creation and transmission. Results suggest that facilities have substantial opportunities to reduce readmissions after psychiatric hospitalization. The association between hospital performance on care coordination process measures and the all-cause readmission measure currently included in the IPFQR program was minimal. The CMS should evaluate whether the IPFQR measures adequately capture compliance with evidence-based processes and desired outcomes.


Preparation and in vivo evaluation of spray dried matrix type controlled-release microparticles of tamsulosin hydrochloride for orally disintegrating tablet.
The objective of this study was to achieve an optimal formulation of spray dried matrix type controlled-release (MTCR) microparticles containing tamsulosin hydrochloride for orally disintegrating tablet. To control the release rate of tamsulosin hydrochloride, Acrylate-methacrylate copolymer (Eudragit(®) L-100 or Eudragit(®) S-100) and ethylcellulose were employed on the composition of MTCR microparticles. Physicochemical properties of MTCR microparticles such as particle size and SEM were characterized. Pharmacokinetic parameters of tamsulosin hydrochloride were evaluated in the rats after oral administration. MTCR microparticles were spherical microparticles of around 10 µm diameter with a corrugated surface. ODTs containing MTCR microparticles were disintegrated within 30 s and MTCR microparticles were able to control the release rate of tamsulosin hydrochloride following Fickian diffusion mechanism. The in vitro release rates of tamsulosin hydrochloride from MTCR microparticles were proportional to the ratio of Acrylate-methacrylate copolymer to ethylcellulose. Moreover, MTCR microparticles retarded the in vivo release rate of tamsulosin hydrochloride without reducing the bioavailability. Our results suggest that MTCR microparticles may be potential oral dosage forms to control the release and to improve the bioavailability of tamsulosin hydrochloride.


A personalized framework for medication treatment management in chronic care.
The ongoing efforts toward continuity of care and the recent advances in information and communication technologies have led to a number of successful personal health systems for the management of chronic care. These systems are mostly focused on monitoring efficiently the patient's medical status at home. This paper aims at extending home care services delivery by introducing a novel framework for monitoring the patient's condition and safety with respect to the medication treatment administered. For this purpose, considering a body area network (BAN) with advanced sensors and a mobile base unit as the central communication hub from the one side, and the clinical environment from the other side, an architecture was developed, offering monitoring patterns definition for the detection of possible adverse drug events and the assessment of medication response, supported by mechanisms enabling bidirectional communication between the BAN and the clinical site. Particular emphasis was given on communication and information flow aspects that have been addressed by defining/adopting appropriate formal information structures as well as the service-oriented architecture paradigm. The proposed framework is illustrated via an application scenario concerning hypertension management.


Male reproductive success of Schistosoma mansoni-infected Biomphalaria glabrata snails.
Infection of Biomphalaria glabrata by Schistosoma mansoni results in a dramatic reduction in the snail's ability to produce eggs. We studied the ability of such parasitically castrated snails to fertilize the eggs of uninfected snails. Pigmented B. glabrata snails (13141 stock) were infected with S. mansoni miracidia and reared individually until they ceased laying eggs. These infected snails were then given the opportunity to mate with uninfected albino (NMRI) snails. Each of the infected snails was paired with a different albino partner each subsequent week. Sperm transfer by the infected snails was evident from the production of pigmented progeny by the uninfected albino snails. Infected snails successfully acted as males for up to 6 wk after parasitic castration had occurred. The duration of allosperm use by uninfected recipients was lengthy, regardless of the infection status of the pigmented sperm donor.


Calories and control: Eating habits, behaviors, and motivations of Generation Z females.
This qualitative phenomenological study explored eating habits, behaviors, and motivations of Gen Z females (born 1996-2002). Ten participants met selection criteria: female aged 18-24 (thus, part of Gen Z), enrolled full-time at the University and able to share experiences. Maximum variation was sought for race/ethnicity, Greek life/sorority involvement and hometown regional area. Semi-structured interviews elicited detailed descriptions of lived experiences. Data consisted of five face-to-face interviews and five Internet-based teleconferenced interviews. Saldana's First Cycle/Second Cycle constant comparative method was used for data analysis. Four themes and four subthemes were identified: Blurred Lines (healthy versus disordered behaviors), Rationalizing Behaviors (Denial), Technological Influences (Social Media; Self-Worth), and Pandemic Influence. Gen Z represents a pivotal population with diverse characteristics. These findings are helpful to health professionals, faculty, academic administrators, and others with vested interests in promoting healthier living, learning and work environments for this population.


[Construction of point mutation plasmids expressing HCV NS3/4A with different secondary structures at amino-terminal and their expressions in Huh 7 cells].
To construct point mutation plasmids expressing HCV NS3/4A with different secondary structures at amino-terminal, and express the constructs in Huh 7 cells. Using pSG5/M-H05-5/4A as the template (A1-1) and primers designed according to the typing criteria, 4 single point mutation plasmids, namely pSG5/M-H05-5(A1-2)/4A(A1-2) (Y56F), pSG5/M-H05-5(B1-1)/4A(B1-1) (L80Q), pSG5/M-H05-5(B2-1)/4A(B2-1) (V51A), and pSG5/M-H05-5(B2-2)/4A(B2-2) (S61A), were constructed. With A1-2, B2-1, and B2-2 as the templates, the leucine to glutamine mutation at position 80 (L80Q) was induced to construct another 3 double point mutation plasmids pSG5/M-H05-5(B1-2)/4A(B1-2), pSG5/M-H05-5(A2-1)/4A(A2-1), and pSG5/M-H05-5(A2-2)/4A(A2-2), respectively. DNA sequencing was performed for confirmation of the mutations. Huh 7 cells were transfected with the constructs using FuGene 6 transfection reagents. Indirect immunofluorescence staining and Western blotting were used to detect the expression of the constructs. Indirect immunofluorescence assay revealed 4 subcellular localization patterns of NS3 protein, including dot-like staining, diffuse staining, doughnut-like staining, and rod-shape staining. Western blotting also demonstrated successful expression of the constructs and weak in cis and in trans NS3 serine protease activities of subtypes A2-1 and B2-1 in comparison with other subtypes. The point mutation plasmids expressing HCV NS3/4A with different secondary structures at amino-terminal are constructed successfully, which provides the basis for further study of different subtypes of HCV.


[High altitude and cardiac disease].
Hypoxia at high altitude is associated with an increase in myocardial work. Cardiovascular effects of high altitude may affect patients suffering from cardiac diseases and raise questions concerning the tolerability of exposure to altitude. In general, altitudes up to 2500 to 3000 meters do not pose a hazard for a patient with stable coronary artery disease and good exercise tolerance. Likewise, a patient suffering from heart failure does not necessarily refrain from these altitude exposures unless exercise capacity and left ventricular ejection fraction are severely impaired. However, cardiac patients have to be informed that symptoms may occur already at lesser efforts at high altitude compared to low altitude. Therefore, patients should be advised to adapt physical activities, particularly during the first 3-4 days of acclimatization. In commercial airplanes pressure conditions are comparable to altitudes of 1500 to 2400 meters. Most cardiac patients tolerante the associated physical stress without difficulty. Contraindications for air travel include unstable cardiac conditions, recent cardiac interventions (2-3 weeks), an uncomplicated myocardial infarction within the last 2 weeks, or a complicated myocardial infarction within the last 6 weeks, respectively. The risk of exposure to altitudes above 3500 meters is not well known for cardiac patients because of a lack of corresponding data. Thus, patients who would like to mountaineer at very high altitudes have to be advised individually based on their physical condition and on the severity of their cardiac illness.


Anatomy, Exposure, and Preparation of Recipient Vessels in Microsurgical Head and Neck Reconstruction.
Successful microvascular reconstruction of head and neck defects requires the ability to safely identify, isolate, and utilize recipient vessels. To date, however, a comprehensive review of the anatomy and techniques relevant to the available anatomic regions has not been undertaken. This review covers the relevant clinical anatomy of the anterior triangle, posterior triangle, submandibular region, intraoral region, preauricular region, chest, and arm, taking particular care to highlight the structures that are crucial to identify while performing each dissection. Finally, a step-by-step technique for safely dissecting the recipient vessels at each site is provided.


Occupational asthma: assessing the risk.
Asthma caused by factors encountered at work can be overlooked. The author describes some of the risk factors and occupational groups involved and how nurses can help to detect cases of asthma caused by materials found at work.


The use of ice for pain associated with chest tube removal.
Every year more than 300,000 patients undergo cardiothoracic surgery, requiring placement of at least one chest tube. Removal of these chest tubes has been described as one of the worst intensive care unit experiences for these patients. Pain associated with chest tube removal (CTR) has been poorly controlled in many surgical patients. The purpose of this experimental study was to ascertain if the application of ice would decrease pain before, during, and after chest tube removal. Fifty postcardiac surgery patients were randomly assigned to two groups. The experimental group received ice therapy before CTR, whereas control subjects received a placebo. Pain intensity and pain distress were measured on a 0-10 numeric rating scale, and pain quality was measured using the McGill Pain Questionnaire-Short Form (MPQ-SF). Differences in pain intensity and pain distress between the experimental and control groups were not significant. A significant change in pain over time was noted in both groups, with pain intensity and distress being most severe during actual chest tube removal. Additionally, patients who received preprocedural pain medication did not differ in their levels of pain intensity or distress. Both groups used all the quality descriptors on the MPQ-SF for the sensory and affective components of pain, with cramping and gnawing as the most frequently chosen words. Continued research with larger samples is encouraged to further evaluate ice and other interventions that can be used to manage pain associated with CTR.


Preparation and characterization of bio-nanocomposite films of agar and silver nanoparticles: laser ablation method.
Silver nanoparticles (AgNPs) were prepared by a laser ablation method and composite films with the AgNPs and agar were prepared by solvent casting method. UV-vis absorbance test and transmission electron microscopy (TEM) analysis results revealed that non-agglomerated spherical AgNPs were formed by the laser ablation method. The surface color of the resulting agar/AgNPs films exhibited the characteristic plasmonic effect of the AgNPs with the maximum absorption peaks of 400-407 nm. X-ray diffraction (XRD) test results also exhibited characteristic AgNPs crystals with diffraction peaks observed at 2θ values of 38.39°, 44.49°, and 64.45°, which were corresponding to (111), (200), and (220) crystallographic planes of face-centered cubic (fcc) silver crystals, respectively. Thermogravimetric analysis (TGA) results showed that thermal stability of the agar/AgNPs composite films was increased by the inclusion of metallic silver. Water vapor barrier properties and surface hydrophobicity of the agar/AgNPs films increased slightly with the increase in AgNPs content but they were not statistically significant (p>0.05), while mechanical strength and stiffness of the composite films decreased slightly (p<0.05). The agar/AgNPs films exhibited distinctive antimicrobial activity against both Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli O157:H7) bacterial pathogens.


Rapid purification and characterization of nucleoside diphosphate kinase isoforms using ATP-sepharose affinity column chromatography.
Nucleoside diphosphate kinases (NDP kinases), products of the nm23 gene, catalyze the transfer of the terminal phosphate group of the nucleoside triphosphate to the corresponding diphosphate and may be involved in tumor metastasis suppression, development, and signal transduction. NDP kinase from various sources including human erythrocytes, rat brain tissue and E. coli strain BL21 transformed with pET3C expression plasmids containing nm23-H1 or nm23-H2, were purified in one step to homogeneity using ATP-sepharose affinity column chromatography. This method was applicable for the purification of various NDP kinases which show the same enzymatic activity and immunodetection, but have various molecular weight and quaternary structures.


Stabilization of homoserine-O-succinyltransferase (MetA) decreases the frequency of persisters in Escherichia coli under stressful conditions.
Bacterial persisters are a small subpopulation of cells that exhibit multi-drug tolerance without genetic changes. Generally, persistence is associated with a dormant state in which the microbial cells are metabolically inactive. The bacterial response to unfavorable environmental conditions (heat, oxidative, acidic stress) induces the accumulation of aggregated proteins and enhances formation of persister cells in Escherichia coli cultures. We have found that methionine supplementation reduced the frequency of persisters at mild (37°C) and elevated (42°C) temperatures, as well as in the presence of acetate. Homoserine-o-succinyltransferase (MetA), the first enzyme in the methionine biosynthetic pathway, is prone to aggregation under many stress conditions, resulting in a methionine limitation in E. coli growth. Overexpression of MetA induced the greatest number of persisters at 42°C, which is correlated to an increased level of aggregated MetA. Substitution of the native metA gene on the E. coli K-12 WE chromosome by a mutant gene encoding the stabilized MetA led to reduction in persisters at the elevated temperature and in the presence of acetate, as well as lower aggregation of the mutated MetA. Decreased persister formation at 42°C was confirmed also in E. coli K-12 W3110 and a fast-growing WErph+ mutant harboring the stabilized MetA. Thus, this is the first study to demonstrate manipulation of persister frequency under stressful conditions by stabilization of a single aggregation-prone protein, MetA.


Type 2 diabetes and obesity metabolic interactions: common factors for breast cancer risk and novel approaches to prevention and therapy.
The objective was to review type 2 diabetes as a risk factor for breast cancer, its influence on tumor aggressiveness and prognosis, and the interactions with obesity. Consideration was given to the responsible biological mechanisms and how these relate to the potential of hypoglycemic agents, notably metformin, as breast cancer chemotherapeutic agents. Most epidemiological studies indicate that type 2 diabetes is a modest positive risk factor for postmenopausal, but not premenopausal, breast cancer; indeed before the menopause it may be associated with a reduced risk. This pattern of differing effects on risk according to menopausal status is well established in obesity; however, although most type 2 diabetics are obese, the relationship with postmenopausal breast cancer does not appear to be a function of the body mass index. We suggest that before menopause the protective effect of obesity may modify any adverse effects of the metabolic changes related to type 2 diabetes. Regardless of menopausal status, obesity is associated with breast cancers that exhibit aggressive biological characteristics at the time of diagnosis and have a poor prognosis; a similar relationship is emerging for type 2 diabetes. The two metabolic disorders share biological mechanisms for their associations with breast cancer, including a direct effect of insulin on breast cancer cell proliferation, increased extraglandular estrogen production and bioavailability, changes in the adipokines, notably adiponectin, and activation of the AMP-activated protein kinase pathway. These mechanistic considerations are consistent with metformin having high potential as a breast cancer chemopreventive and therapeutic agent.


Fizeau digital interferometry with a diffraction-generated spherical wave for testing focusing optics.
A sufficiently small, reflective spherical particle at the focus of a test lens can replace the spherical autostigmatizing mirror in Fizeau digital interferometry, which is used for testing focusing optics. This small particle serves as a secondary coherent point source in the test arm and generates by diffraction a spherical wave that is used to produce the test beam. The relationships between the diffraction-generated wave and the size and shape of the diffracting element are discussed and also used to properly select the diffracting element. Experimental results confirm that the new technique has high accuracy.


Zac1 and the Imprinted Gene Network program juvenile NAFLD in response to maternal metabolic syndrome.
Within the next decade, NAFLD is predicted to become the most prevalent cause of childhood liver failure in developed countries. Predisposition to juvenile NAFLD can be programmed during early life in response to maternal metabolic syndrome (MetS), but the underlying mechanisms are poorly understood. We hypothesized that imprinted genes, defined by expression from a single parental allele, play a key role in maternal MetS-induced NAFLD, due to their susceptibility to environmental stressors and their functions in liver homeostasis. We aimed to test this hypothesis and determine the critical periods of susceptibility to maternal MetS. We established a mouse model to compare the effects of MetS during prenatal and postnatal development on NAFLD. Postnatal but not prenatal MetS exposure is associated with histological, biochemical, and molecular signatures of hepatic steatosis and fibrosis in juvenile mice. Using RNA sequencing, we show that the Imprinted Gene Network (IGN), including its regulator Zac1, is up-regulated and overrepresented among differentially expressed genes, consistent with a role in maternal MetS-induced NAFLD. In support of this, activation of the IGN in cultured hepatoma cells by overexpressing Zac1 is sufficient to induce signatures of profibrogenic transformation. Using chromatin immunoprecipitation, we demonstrate that Zac1 binds the TGF-β1 and COL6A2 promoters, forming a direct pathway between imprinted genes and well-characterized pathophysiological mechanisms of NAFLD. Finally, we show that hepatocyte-specific overexpression of Zac1 is sufficient to drive fibrosis in vivo. Our findings identify a pathway linking maternal MetS exposure during postnatal development to the programming of juvenile NAFLD, and provide support for the hypothesis that imprinted genes play a central role in metabolic disease programming.


Evidence for a matrix metalloproteinase induction/activation system in arterial vasculature and decreased synthesis and activity in diabetes.
Pathological remodeling characterized by extracellular matrix (ECM) deposition contributes to the diabetic vascular complications. Matrix metalloproteinases (MMPs) regulate ECM turnover. However, the expression profile of the MMP system in diabetic human tissue remains unknown. The objectives of this study were 1) to identify a local MMP induction/activation system that exists in arterial vasculature and 2) to determine how the MMP system may be altered in diabetes. Internal mammary artery specimens were obtained from patients who did (n = 14) and did not (n = 14) have diabetes and were undergoing coronary artery bypass grafting surgery. ECM inducer protein (EMMPRIN); membrane-type MMP (MT-MMP); and MMP-1, -2, and -9 were quantified by immunoblotting and densitometric scanning (pixels). Pro-MMP-1 and MMP-2 levels were decreased from 952 +/- 120 and 1,081 +/- 508 pixels, respectively, in nondiabetic tissue to 398 +/- 62 and 249 +/- 42 pixels in the diabetic tissue (P < 0.05). Both EMMPRIN and MT-MMP expression and total MMP activity were decreased by twofold in diabetic patients (P < 0.05). These results demonstrated for the first time that an MMP induction and activation system exists in human arterial vasculature and that it is downregulated in diabetes. Decreased MMP activity may contribute to increased collagen deposition and pathological remodeling in diabetes.


Phenotypic and genetic analysis of the cerebellar mutant tmgc26, a new ENU-induced ROR-alpha allele.
ROR-alpha is an orphan nuclear receptor, inactivation of which cell-autonomously blocks differentiation of cerebellar Purkinje cells with a secondary loss of granule neurons. As part of our ENU mutagenesis screen we isolated the recessive tmgc26 mouse mutant, characterized by early-onset progressive ataxia, cerebellar degeneration and juvenile lethality. Detailed analysis of the tmgc26-/- cerebella revealed Purkinje cell and granule cell abnormalities, and defects in molecular layer interneurons and radial glia. Chimera studies suggested a cell-autonomous effect of the tmgc26 mutation in Purkinje cells and molecular layer interneurons, and a non-cell-autonomous effect in granule cells. The mutation was mapped to a 13-Mb interval on chromosome 9, a region that contains the ROR-alpha gene. Sequencing of genomic DNA revealed a T-to-A transition in exon 5 of the ROR-alpha gene, resulting in a nonsense mutation C257X and severe truncation of the ROR-alpha protein. Together, our data identify new roles for ROR-alpha in molecular layer interneurons and radial glia development and suggest tmgc26 as a novel ROR-alpha allele that may be used to further delineate the molecular mechanisms of ROR-alpha action.


Antagonistic Control of Genetic Circuit Performance for Rapid Analysis of Targeted Enzyme Activity in Living Cells.
Genetic circuits have been developed for quantitative measurement of enzyme activity, metabolic engineering of strain development, and dynamic regulation of microbial cells. A genetic circuit consists of several bio-elements, including enzymes and regulatory cassettes, that can generate the desired output signal, which is then used as a precise criterion for enzyme screening and engineering. Antagonists and inhibitors are small molecules with inhibitory effects on regulators and enzymes, respectively. In this study, an antagonist and an inhibitor were applied to a genetic circuit for a dynamic detection range. We developed a genetic circuit relying on regulators and enzymes, allowing for straightforward control of its output signal without additional genetic modification. We used para-nitrophenol and alanine as an antagonist of DmpR and inhibitor of tyrosine phenol-lyase, respectively. We show that the antagonist resets the detection range of the genetic circuit similarly to a resistor in an electrical logic circuit. These biological resistors in genetic circuits can be used as a rapid and precise controller of variable outputs with minimal circuit configuration.


Clinical significance of the tumor microenvironment in non-small cell lung cancer.
Several biomarkers have been reported as predictors of survival and recurrence in non-small cell lung cancer. Recently, several groups have demonstrated that the immune microenvironment of the primary tumors is a prognostic factor. These "immunological biomarkers" in the tumor microenvironment are useful predictors of prognosis as well as promising targets for novel therapeutic approaches. Especially, tumor-infiltrating Treg cells are a powerful immunological biomarker, and possible mechanisms involved in the induction of tumor-infiltrating Treg cells are the expression of Cox-2, IL-12Rβ2 or the lack of IL7R on the tumor cells. These findings may pave the way for individualized immunomodulatory therapies to deplete tumor-infiltrating Treg cells from the tumor microenvironment.


The effect of moderate and excessive alcohol use on the course and outcome of patients with bipolar disorders: a prospective cohort study.
Comorbid alcohol use disorders (AUDs) are frequently associated with negative effects on course and outcome of bipolar disorder. This prospective cohort study assessed the effect of actual alcohol use (no, moderate, and excessive) on the course and outcome of patients with bipolar disorders. Between June 2003 and November 2005, 137 outpatients (aged 23-68 years) with DSM-IV-diagnosed bipolar I (66%) or II (34%) disorder rated their mood and the number of alcohol units consumed daily for a period up to 52 weeks with the National Institute of Mental Health Self-Rating Prospective Life-Chart Method (LCM). At baseline, the Structured Clinical Interview for DSM-IV was administrated, and demographic, social, and clinical characteristics were obtained. At monthly visits, the Clinical Global Impressions Scale-Bipolar Version (CGI-BP), the Global Assessment of Functioning (GAF) scale, and the Medical Outcomes Study 36-Item Short-Form Health Survey (MOS-SF-36) were rated. Based on the alcohol use in the first 4 weeks of follow-up, patients were assigned to 1 of 3 groups: no/incidental, moderate, or excessive alcohol use. None of the sociodemographic and clinical characteristics at baseline were significantly different between the 3 drinking groups, with the exception of-and as a consequence of the group assignment-the prevalence of lifetime and current diagnosis of AUD. Also, no differences between the 3 drinking groups were found on any of the clinical outcome variables, ie, number of days ill (depressed, hypomanic/manic, and total); severity of depression, mania, and overall bipolar illness (LCM); GAF score; CGI-BP (depression, mania, and overall); and all the subscales of the MOS-SF-36. Also, the number of episodes according to DSM-IV and the Leapfrog method showed no significant differences between the drinking groups. In this sample of patients and with the sensitive measurement of mood and drinking status over a full year, we could not confirm the findings of other studies indicating a negative effect of excessive alcohol use on the course of bipolar illness. This study found that neither moderate nor excessive use of alcohol has a negative effect on the course and outcome of bipolar illness. Possible explanations for these findings are discussed.


A meta-analysis of studies investigating the effects of occupational lead exposure on thyroid hormones.
Investigations of the effects of occupational exposure to lead on the concentrations of thyroid hormones in the blood have not produced consistent results. A meta-analysis was performed to assess the effect of occupational exposure to lead on thyroid hormone concentrations using the results from published studies. Group means from studies of the thyroid function of persons occupationally exposed to lead were used in a meta-analysis. Differences between the control and exposed groups, and the slopes between thyroid hormone concentrations and log10 blood lead concentrations or duration of exposure to lead were estimated using mixed models. The hormones analyzed were thyroid stimulating hormone, total and free thyroxine, and total and free triiodothyronine. No differences in mean thyroid hormone concentrations were found between the exposed and control groups. No relationships were found between blood lead or the duration of exposure to lead and thyroid hormone concentrations. The results of the analysis do not provide evidence for an effect of occupational lead exposure on thyroid function in men. Am. J. Ind. Med. 59:583-590, 2016. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.


Three-dimensional printing in paediatric orthopaedic surgery.
Three-dimensional (3D) printing is a rapidly evolving and promising field to improve outcomes of orthopaedic surgery. The use of patient-specific 3D-printed models is specifically interesting in paediatric orthopaedic surgery, as limb deformity corrections often require an individual 3D treatment. In this editorial, various operative applications of 3D printing in paediatric orthopaedic surgery are discussed. The technical aspects and the imaging acquisition with computed tomography and magnetic resonance imaging are outlined. Next, there is a focus on the intraoperative applications of 3D printing during paediatric orthopaedic surgical procedures. An overview of various upper and lower limb deformities in paediatrics is given, in which 3D printing is already implemented, including post-traumatic forearm corrections and proximal femoral osteotomies. The use of patient-specific instrumentation (PSI) or guiding templates during the surgical procedure shows to be promising in reducing operation time, intraoperative haemorrhage and radiation exposure. Moreover, 3D-printed models for the use of PSI or patient-specific navigation templates are promising in improving the accuracy of complex limb deformity surgery in children. Lastly, the future of 3D printing in paediatric orthopaedics extends beyond the intraoperative applications; various other medical applications include 3D casting and prosthetic limb replacement. In conclusion, 3D printing opportunities are numerous, and the fast developments are exciting, but more evidence is required to prove its superiority over conventional paediatric orthopaedic surgery.


Laser-induced regeneration of cartilage.
Laser radiation provides a means to control the fields of temperature and thermo mechanical stress, mass transfer, and modification of fine structure of the cartilage matrix. The aim of this outlook paper is to review physical and biological aspects of laser-induced regeneration of cartilage and to discuss the possibilities and prospects of its clinical applications. The problems and the pathways of tissue regeneration, the types and features of cartilage will be introduced first. Then we will review various actual and prospective approaches for cartilage repair; consider possible mechanisms of laser-induced regeneration. Finally, we present the results in laser regeneration of joints and spine disks cartilages and discuss some future applications of lasers in regenerative medicine.


Effect of the Kallikrein Inhibitor Aprotinin on Myocardial Ischemia and Necrosis in Man.
The effect of administration of aprotinin, a serine esterase inhibitor capable of inactivating kallikrein, on the extent and severity of acute myocardial ischemic injury and subsequent necrosis, was studied in 25 patients. Another group of 25 patients who did not receive aprotinin served as a control group.We administered 100,000 kallikrein inhibitor units (KIU) of aprotinin as a bolus dose, followed by a continuous infusion (4 ml/min) that contained 10,000 KIU/kg in 240 ml of dextrose/water solution, to all 25 patients admitted to the hospital within 30 to 60 minutes after the onset of acute myocardial ischemia.To measure the effect of aprotinin, three parameters were studied; the sum of S-T segment elevations (ΣST), the development of Q waves, and the prediction of infarct size by measuring the disappearance rate of creatine phosphokinase(MB CPK isoenzyme).The average ΣST in the treated group decreased from 40.5 ± 7.00 mv to 12.95 ±4.60 mv (P < 0.01); in contrast the control group's ΣST did not change significantly, from 54.25 ± 8.02 to 51.7 ± 6.8. Deeper Q waves evolved in the control group compared to the treated group: ΔQ (6 hours) = 1.0 ST (15 min) +1.19 (25 patients, r = 0.78); and in the treated group ΔQ (6 hours) = 0.66 ST (15min) + 0.91 (25 patients, r = 0.65) (P < 0.025).In the control group the estimated infarct size was 57.4 ± 4 CPK-gram equivalents(CPK-g-Eq). There was significantly less damage in the treated group: 19 ± 2 CPK-g-Eq (P < 0.01). Thus we conclude that aprotinin diminishes myocardial damage.


Gastro-Resistant Insulin Receptor-Binding Peptide from Momordica charantia Improved the Glucose Tolerance in Streptozotocin-Induced Diabetic Mice via Insulin Receptor Signaling Pathway.
Momordica charantia is a commonly used food and has been used for the management of diabetes. Our previous study has identified an insulin receptor (IR)-binding protein (mcIRBP) from Momordica charantia. Here we identified the gastro-resistant hypoglycemic bioactive peptides from protease-digested mcIRBP. By in vitro digestion and IR kinase activity assay, we found that a 9-amino-acid-residue peptide, mcIRBP-9, was a gastro-resistant peptide that enhanced IR kinase activities. mcIRBP-9 activated IR signaling transduction pathway, which resulted in the phosphorylation of IR, the translocation of glucose transporter 4, and the uptake of glucose in cells. Intraperitoneal and oral administration of mcIRBP-9 stimulated the glucose clearance by 30.91 ± 0.39% and 32.09 ± 0.38%, respectively, in streptozotocin-induced diabetic mice. Moreover, a pilot study showed that daily ingestion of mcIRBP-9 for 30 days decreased the fasting blood glucose levels and glycated hemoglobin (HbA1c) levels by 23.62 ± 6.14% and 24.06 ± 1.53%, respectively. In conclusion, mcIRBP-9 is a unique gastro-resistant bioactive peptide generated after the digestion of mcIRBP. Furthermore, oral administration of mcIRBP-9 improves both the glucose tolerance and the HbA1c levels in diabetic mice via targeting IR signaling transduction pathway.


Association of free fatty acids with the insulin-resistant state but not with central obesity in individuals from Venezuela.
Individuals with insulin resistance (IR) usually have upper body obesity phenotype, often accompanied by an increase in plasma free fatty acids (FFA). Since the Venezuelan population has a high frequency of IR and central obesity, the purpose of this work was to determine FFA levels in 47 Venezuelan individuals, men and women, 24-58 years old, and analyze their relationship with central obesity and parameters of carbohydrate and lipid metabolism. Basal concentrations of TG, total cholesterol, LDL-C, and HDL-C were measured, and FFA, glucose and insulin, at basal state and at different times after a glucose load. Eighteen individuals presented insulin resistance (HOMA-IR > 2.7) and 29 were non-insulin resistant (non-IR). Insulin resistant individuals (IR) had higher waist circumference, BMI and basal concentrations of FFA than the non-IR. No differences were observed in skin folds and other basal lipids studied. The increased FFA seemed to be related to the IR associated to BMI and not to central obesity, since the difference between IR and non-IR disappeared when they were matched for waist circumference. After a glucose load, FFA decreased in both groups, but remained significantly elevated in IR subjects. This effect disappeared after matching for BMI or waist circumference, inferring that it was independent of anthropometries. FFA were positively associated with HOMA-IR, glucose and TG levels; however, there was.no association with BMI or waist circumference. These findings, and the lack of elements to support the presence of hepatic IR, common to increased visceral lipolysis, might suggest that the IR present in the obese individuals studied, might be due to an increase in subcutaneous fat.


Acute episodic hemolysis in the African black rhinoceros as an analogue of human glucose-6-phosphate dehydrogenase deficiency.
Sudden episodes of massive hemolysis have become the most common cause of death among captive black rhinoceroses, and there is evidence that they occur in the wild as well. We have observed radically unique enzyme and metabolite profiles in normal rhinoceros erythrocytes compared to humans and other mammals, including marked deficiencies of intracellular adenosine triphosphate (ATP), catalase, adenosine deaminase, and other enzymes involved in glycolysis, glutathione cycling, and nucleotide metabolism. Minimal concentrations of ATP appear to impair effective acceleration of hexosemonophosphate shunt activity in response to oxidants by restricting substrate generation at the hexokinase step. Antioxidant defenses are further compromised by catalase deficiency, which may be a general characteristic of rhinoceros erythrocytes, perhaps related to the common occurrence of severe mucocutaneous ulcerative disease. It is proposed that erythrocyte ATP deficiency in rhinoceroses may be an evolutionary adaptation conferring selective advantage against common hemic parasites, comparable to the role of human glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in falciparum malaria.


Assessment of oral health status among uncontrolled diabetic mellitus patients in Tunisia.
To assess oral health status of Tunisian diabetic mellitus patients. We carried a cross-sectional study among 100 uncontrolled diabetic mellitus patients. An oral health survey was developed to collect their socio-demographic characteristics and to assess their oral health behavior. Each patient benefited of an oral examination. Clinical and biological data were collected from medical file of patients. In terms of number of brushings, 44% of diabetic mellitus patients brushed their teeth at least twice a day, 25% brushed once a day, 10% occasionally and 21% did not used to brush their teeth. The last visit to a dentist dated since less than a year ago for 48% of participants and 8% reported that they hadn't never visit a dentist. Oral hygiene and the rate of dental consultation were correlated with gender but not with glycemic control. Only 17% had complete dentition, 75% were partially dentate and 8% were completely edentulous. More than half of the diabetics had at least 5 missing teeth. About 67% had had experienced tooth decay. The presence of tooth decay was not correlated with glycated hemoglobin (HbA1c). The examining dentist estimated that nine out of ten patients required dental care. The oral health status of diabetic mellitus patients in Tunisia was bad. The need for dental care was real and high but often overlooked. The establishment of a national oral health care policy seems to be a national priority today.


Transendothelial glucose transport is not restricted by extracellular hyperglycaemia.
Endothelial cells are routinely exposed to elevated glucose concentrations post-prandially in healthy individuals and permanently in patients with metabolic syndrome and diabetes, and so we assessed their sugar transport capabilities in response to high glucose. In human umbilical vein (HUVEC), saphenous vein, microdermal vessels and aorta, GLUT1 (SLC2A1), GLUT3 (SLC2A3), GLUT6 (SLC2A6), and in microdermal vessels also GLUT12 (SLC2A12), were the main glucose transporters as assessed by mRNA, with no fructose transporters nor SGLT1 (SLC5A1). Uptake of 14C-fructose was negligible. GLUT1 and GLUT3 proteins were detected in all cell types and were responsible for ~60% glucose uptake in HUVEC, where both GLUT1 and GLUT3, but not GLUT6 siRNA knock-down, reduced the transport. Under shear conditions, GLUT1 protein decreased, GLUT3 increased, and 14C-deoxy-glucose uptake was attenuated. In high glucose, lipid storage was increased, cell numbers were lower, 14C-deoxy-glucose uptake decreased owing to attenuated GLUT3 protein and less surface GLUT1, and trans-endothelial transport of glucose increased due to cell layer permeability changes. We conclude that glucose transport by endothelial cells is relatively resistant to effects of elevated glucose. Cells would continue to supply it to the underlying tissues at a rate proportional to the blood glucose concentration, independent of insulin or fructose.


[Occupational exposure to hand-transmitted vibration in Poland].
Occupational exposure to hand transmitted vibration may cause disorders in upper extremities known as hand-arm vibration syndrome. Therefore it is essential to know the sources of vibration, occupational groups exposed to vibration and the number of exposed workers. The aim of the study was to estimate the number of men and women exposed to hand-transmitted vibration in Poland. The completed questionnaires were obtained from 265 (80%) sanitary inspection stations. They included questions on: the name of workplaces, the name and the type of vibration sources, workers' gender, the number of workers exposed to vibration, indicating the extent of exposure measured against the three threshold limit values (< 0.5 TLV; 0.5 < TLV < 1 and > 1 TLV), and the number of workers exposed to hand-transmitted vibration not documented by measurements in a particular workplaces, indicating one of the three possible kinds of exposure (occasional, periodical and constant). The questionnaire data were based on measurements and analyses performed in 1997-2000. The results of the study showed that vibrating tools used by grinders, fitters, locksmiths, rammers, road workers, carpenters and smiths proved to be the most frequent sources of hand-transmitted vibration. It was revealed that 78.6% of operators of these tools were exposed to vibration exceeding 1 TLV. The study also indicated that 17,000 workers, including 1700 women, were exposed to vibration exceeding the threshold limit values.


Ammonium tetrathiomolybdate treatment targets the copper transporter ATP7A and enhances sensitivity of breast cancer to cisplatin.
Cisplatin is an effective breast cancer drug but resistance often develops over prolonged chemotherapy. Therefore, we performed a candidate approach RNAi screen in combination with cisplatin treatment to identify molecular pathways conferring survival advantages. The screen identified ATP7A as a therapeutic target. ATP7A is a copper ATPase transporter responsible for intercellular movement and sequestering of cisplatin. Pharmaceutical replacement for ATP7A by ammonium tetrathiomolybdate (TM) enhanced cisplatin treatment in breast cancer cells. Allograft and xenograft models in athymic nude mice treated with cisplatin/TM exhibited retarded tumor growth, reduced accumulation of cancer stem cells and decreased cell proliferation as compared to mono-treatment with cisplatin or TM. Cisplatin/TM treatment of cisplatin-resistant tumors reduced ATP7A protein levels, attenuated cisplatin sequestering by ATP7A, increased nuclear availability of cisplatin, and subsequently enhanced DNA damage and apoptosis. Microarray analysis of gene ontology pathways that responded uniquely to cisplatin/TM double treatment depicted changes in cell cycle regulation, specifically in the G1/S transition. These findings offer the potential to combat platinum-resistant tumors and sensitize patients to conventional breast cancer treatment by identifying and targeting the resistant tumors' unique molecular adaptations.


Seeing shapes in seemingly random spatial patterns: Fractal analysis of Rorschach inkblots.
Rorschach inkblots have had a striking impact on the worlds of art and science because of the remarkable variety of associations with recognizable and namable objects they induce. Originally adopted as a projective psychological tool to probe mental health, psychologists and artists have more recently interpreted the variety of induced images simply as a signature of the observers' creativity. Here we analyze the relationship between the spatial scaling parameters of the inkblot patterns and the number of induced associations, and suggest that the perceived images are induced by the fractal characteristics of the blot edges. We discuss how this relationship explains the frequent observation of images in natural scenery.


Molecular analysis of colonic transformation in the ileum after total colectomy in rats.
Colonic transformation is thought to be a phenotypic alteration in the ileum after total colectomy (TC) but has not been well addressed at the molecular level. We previously demonstrated roles of aldosterone in enhancement of ileal sodium absorption after TC. However, the significance of aldosterone in intestinal adaptation has been unknown. Rat epithelial gene expression was compared between the ileum and distal colon by complementary DNA microarray. Genes were categorized into "colonic," "common," and "ileal" genes according to signal intensity. Ileal gene expressions in (1) the control and TC rats and (2) the control and aldosterone-infused rats were compared to detect altered genes or to assess the role of aldosterone in intestinal adaptation. Differential expression of MUC3 and lysozyme was confirmed by quantitative reverse transcriptase-polymerase chain reaction. A total of 6109 genes were categorized into "colonic," "common," or "ileal" gene pools. A comparison of the control and TC rats yielded 82 and 91 genes that were induced and suppressed in the ileum after TC, respectively. Thirty-five percent of them were associated with colonlike transformation (ie, the induction of colonic genes or the suppression of ileal genes). The expressions of MUC3 and lysozyme messenger RNAs were enhanced significantly in the TC ileum, compared with the control. In comparison, aldosterone infusion modulated a total of only 21 genes in the ileum. Those data demonstrate that altered gene expression after TC is, in part, characterized by colonlike transformation. Furthermore, circulating aldosterone appears to play a part in the altering and/or adapting of gene expression in intestinal epithelium.


Efficacy of D5F3 IHC for detecting ALK gene rearrangement in NSCLC patients: a systematic review and meta-analysis.
We conducted a pooled analysis comparing the efficacy of an immunohistochemistry (IHC) assay using the D5F3 antibody with that of fluorescence in situ hybridization (FISH) for detecting ALK gene rearrangement in NSCLC patients. A total of 32 studies involving 5805 samples were included in this review. Pooled sensitivity for D5F3 IHC was 0.97 (95%CI: 0.93-0.98), specificity was 0.99 (95%CI: 0.98-1.00), PLR was 119.20 (95%CI: 57.79-245.89), NLR was 0.03 (95%CI: 0.02-0.07), DOR was 3526.66 (95%CI: 1344.71-9249.03), and AUROC was 1.00 (95%CI: 0.99-1.00). Meta-regression revealed that specimen type was a source of heterogeneity for specificity, and specimen type and FISH signal distance were sources of heterogeneity in the joint model. Subgroup analysis revealed that sensitivity and specificity were higher when the FISH signal distance standard was ≥ 2 than when it was ≥ 1. Sensitivity was higher for tumor specimens than for cell specimens; specificity was higher for cell specimens than for tumor specimens. In conclusion, the D5F3 IHC assay was nearly as effective as FISH for detection of ALK gene rearrangement in NSCLC patients.


Number of Examined Lymph Nodes as a Risk Factor for Recurrence in pT1N+ or pT2-3N0 Gastric Cancer.
Japanese Gastric Cancer Treatment Guidelines do not recommend adjuvant chemotherapy after radical gastrectomy for pathological stage (p) T1N+ or pT2~3N0 gastric cancer. However, some patients experience disease recurrence. This study aimed to identify the risk factors for recurrence in pT1N+ or pT2-3N0 gastric cancer. The study included 157 patients with diagnosed pT1N+ or pT2-3N0 gastric cancer who underwent radical gastrectomy at our institution between January 2001 and December 2020. Clinicopathological data and surgical data were obtained. Independent prognostic factors were analyzed using a Cox proportional hazards regression model. Thirteen patients (8.3%) experienced disease recurrence. Multivariate analysis revealed that the number of examined lymph nodes was an independent prognostic factor for recurrence-free survival (hazard ratio=10.90; 95% confidence interval=1.39-85.86; p=0.023). The group with ≤35 examined lymph nodes had significantly worse recurrence-free survival compared with the group with ≥36 examined lymph nodes (80.7% versus 98.7%; p=0.0005). The number of examined lymph nodes (≤35) was an independent risk factor for recurrence after radical gastrectomy with pT1N+ or pT2-3N0 gastric cancer.


AT1 receptors mediate angiotensin II-induced release of nitric oxide in afferent arterioles.
Recent studies have indicated that angiotensin II (Ang II) possibly activates the nitric oxide (NO) system. We investigated the role of AT receptor subtypes (AT-R) in mediating the Ang II-induced NO release in afferent arterioles (Af) of mice. Isolated Af of mice were perfused, and the isotonic contraction measured. Further, NO release was determined using DAF-FM, a fluorescence indicator for NO. Moreover, we qualitatively assessed the expression of AT-R at the mRNA level using reverse transcription-polymerase chain reaction (RT-PCR). Ang II reduced luminal diameters dose dependently (67.3 +/- 6.3% at 10(-6) mol/L). Inhibition of AT2-R with PD123.319 did not change the Ang II contractile response. AT1-R blockade with ZD7155 inhibited contraction. Stimulation of AT2-R during AT1-R inhibition with ZD7155, and preconstriction with norepinephrine (NE) had no influence on the diameter. Drug application via the perfusion pipette changed flow and pressure, and enhanced NO fluorescence by DeltaF = 4.0 +/- 0.4% (N= 14, background). Luminal application of Ang II (10(-7) mol/L) increased the NO fluorescence by DeltaF = 9.9 +/- 1.2% (N= 8). AT1-R blockade blunted the increase to background levels (DeltaF to 4.0 +/- 0.3%, N= 6, P < 0.05), but AT2-R blockade did not (8.1 +/- 0.9%, N= 9). L-NAME nearly abolished the Ang II effect on the NO fluorescence (DeltaF = 1.6 +/- 0.5% (N= 8). NE did not increase NO release beyond the background levels. RT-PCR showed expression of both AT1-R and AT2-R. The results indicate an Ang II-induced NO release in Af of mice, which is mediated by AT1-R. Thus, Ang II balances its own constrictor action in Af. This control mechanism is very important in view of high renin and angiotensin II concentration in the juxtaglomerular apparatus.


Brain metastases.
Brain metastases are an increasingly common complication in patients with systemic cancer. The optimal treatment for each patient depends on careful evaluation of several factors: the location, size, and number of brain metastases; the patient's age, general condition, and neurologic status; and the extent of systemic cancer to name a few. For patients with a single brain metastasis and limited systemic disease, the standard treatment is surgical resection followed by whole brain radiation therapy. In patients with a small, single metastasis, stereotactic radiosurgery is probably comparable to surgery. Patients with several metastases (up to three) and controlled systemic disease can be treated with whole-brain radiation and stereotactic radiosurgery. Patients with multiple metastases (more than three) are generally treated with whole-brain radiation alone. Radiosurgery is effective in treating patients with a limited number of recurrent brain metastases and stable systemic diseases. Surgery may have a role in patients with a large symptomatic recurrent lesion producing mass effect. Reirradiation and chemotherapy may have a limited role in patients with multiple recurrent metastases.


Considerations of "Combined Probability of Injury" in the next-generation USA frontal NCAP.
The numerical basis for assigning star ratings in the next-generation USA New Car Assessment Program (NCAP) for frontal impacts was assessed. That basis, the Combined Probability of Injury, or CPI, is the probability of an occupant sustaining an injury to any of the specified body regions. For an NCAP test, a CPI value is computed by (a) using risk curves to convert body-region responses from a test dummy into body-region risks and (b) using a theoretical, overarching CPI equation to convert those separate body-region risks into a single CPI value. Though the general concept of applying a CPI equation to assign star ratings has existed since 1994, there will be numerous changes to the 2011 frontal NCAP: there will be two additional body regions (n = 4 vs. 2), the injury probabilities will be evaluated for lower-severity (more likely) injury levels, and some of the occupant responses will change. These changes could yield more disperse CPIs that could yield more disperse ratings. However, the reasons for this increased dispersion should be consistent with real-world findings. Related assessments were the topic of this two-part study, focused on drivers. In Part 1, the CPI equation was assessed without applying risk curves. Specifically, field injury probabilities for the four body regions were used as inputs to the CPI equation, and the resulting equation-produced CPIs were compared with the field CPIs. In Part 2, subject to analyses of test dummy responses from recent NCAP tests, the effect of risk curve choice on CPIs was assessed. Specifically, dispersion statistics were compared for CPIs based on various underlying risk curves applied to data from 2001-2005 model year vehicles (n = 183). From Part 1, the theoretical CPI equation for four body regions demonstrated acceptable fidelity when provided field injury rates (R(2)= 0.92), with the equation-based CPIs being approximately 12 percent lower than those of ideal correlation. From Part 2, the 2011 NCAP protocol (i.e., application of a four-body-region CPI equation whose inputs were from risk curves) generally increased both the CPIs and their dispersion relative to the current NCAP protocol. However, the CPIs generally increased due to an emphasis on neck injury-an emphasis not observed in real-world crashes. Subject to alternative risk curves for the neck and chest, again there was increased dispersion of the CPIs, but the unrealistic emphasis on the neck was eliminated. However, risk estimates for the knee/thigh/hip (KTH) for NCAP-type events remained understated and did not fall within the confidence bands of the field data. Accordingly, KTH risk estimation is an area for future research.


Analysis of 6R- and 6S-tetrahydrobiopterin and other pterins by reversed-phase ion-pair liquid-chromatography with fluorimetric detection by post-column sodium nitrite oxidation.
A rapid and sensitive reversed-phase ion-pair liquid chromatographic system with fluorimetric detection by post-column sodium nitrite oxidation was established for measuring six pterin compounds (6R-5,6,7,8-tetrahydrobiopterin, 6S,5,6,7,8-tetrahydrobiopterin, 7,8-dihydrobiopterin, biopterin, pterin and D-neopterin). The coefficients of variation for these pterins were 0.705-3.714%, and the minimum detectable amount was ca. 10-20 pg at a signal-to-noise ratio of 3. A linear detector response was also verified. The concentrations of the pterin compounds in rat tissues were measured by the described method. Furthermore, by means of brain microdialysis, the output of pterin compounds from rat striatum was detected. Therefore, these results demonstrate that this system can be applied to analyses not only of various rat tissues but also of dialysates collected in vivo.


The significance of bleeding patterns in Norplant implant users.
The purpose of this study was to examine the bleeding patterns of 234 Norplant users during 5 years of use and to identify the bleeding patterns of users who conceived. During the first year of use, 26.6% of users had regular bleeding cycles, 66.3% had irregular cycles, and 7.1% were amenorrheic. By the fifth year of use, 62.5% of users had regular cycles, 37.5% had irregular cycles, and none had amenorrhea. Of the ten users who became pregnant, eight had regular menstrual cycles in the 6 months before the diagnosis of pregnancy, one had an irregular pattern, and one did not keep a bleeding record. None had amenorrhea. The 5-years cumulative pregnancy rate for patients with regular cycles was 17.4%; this was significantly higher (P less than .05) than the 5-year cumulative rates of 4.4% in users with irregular cycles and 0% in users with amenorrhea. This study indicates that during the first year of Norplant use, only 26.6% of users have regular cycles, but after the first year, 50-60% of users develop regular cycles. The bleeding patterns of women using Norplant improve after the first year of use, and those with regular cycles are at greatest risk for method failure.


Malignant insulinoma with hepatic and pulmonary metastases associated with primary hyperparathyroidism. Case report and review of the literature.
Malignant insulinomas are rare tumors (10% of insulinomas) that often present as mnulticentric macro nodules with multiple liver metastases before diagnosis. We report the case of a 55 year old female with a medical history of severe hypoglycemic attacks for two months. Blood tests showed a decreased value of glycemia (30 mg/dl) associated with increased insulin level (l6 microU/ml) and an increased glycemia/insulinemia ratio of 1.87 supporting the diagnosis of insulinoma. Abdominal CT showed a 1.5 cm mass localized in the head of the pancreas with disseminated hepatic tumors, confirmed as neuroendocrine metastases by biopsy (which proved the presence of a malignant insulinoma). Primary hyperparathyroidism was diagnosed based on mild elevation of calcium (10.4 mg/dl) associated with a high level of PTH (71.2 pg/ml). The coexistence of the two endocrinopathies suggested the presence of type 1 multiple endocrine neoplasia (MEN 1). Because of multiple hepatic masses and liver function impairment, surgery and hepatic artery embolization were not performed. Somatostatin analog therapy was started with symptomatic control in the beginning, but rapid loss of beneficial effect. Finally, systemic chemotherapy with doxorubicin was administered, but the disease was progressive and after three months we decided to stop it. The patient died at home after one month, probably in hypoglycemic coma.


Using polypropylene needle punch nonwoven sorbents as the interceptor for oil in static and dynamic water experiments.
This work was to determine the impact of pore size and thickness of sorbents besides the influence of oil properties on the performance of sorbents used as an interceptor barrier for oils in Static and dynamic water experiments. polypropylene needle punch sorbents were used. SEM test showed the porous structure of sorbents. Oil sorption test investigated that with sorbents had same thickness N1 absorbed oil 8.89 g/g of soybean oil and 7.15 g/g of motor oil. Meanwhile, sorbents with the same pore size, N6 absorbed oil 6.11 g/g of soybean oil and 5.13 g/g of motor oil. All sorbents showed a retention rate of over 75% after 24 h of dripping. Dynamic oil spreading revealed that smaller pore size and higher thickness exhibited higher height wicking. The static performance experiment showed motor oil and soybean oil started to leak at 38 and 32 min for N1, then prolonged with reducing pore size with intercepting efficiency around 70% after 600 min for motor oil and 540 min for soybean oil. In comparison, The motor oil and soybean oil started to leak at 49 and 40 min for N4 and then prolonged with increased thickness with intercepting efficiency around 70% after 480 min for motor oil and 360 min for soybean oil. Under the dynamic experiment, oils spilled quicker with initial leakage of sorbents N3 and N6 at rate flow 55.65 ml/s for motor oil and soybean oil were (41 min, 36 min) and (50 min, 41 min) while intercepting efficiency was (99%, 98.40%) and (99.33%, 98.40%).


Direct methoxypyridine functionalization approach to magellanine-type Lycopodium alkaloids.
A concise enantioselective approach to the tetracyclic core of the magellanine-type Lycopodium alkaloids is reported. Key to this approach is the use of the Hajos-Parrish reaction to set a challenging quaternary stereocenter, thereby guiding the stereoselectivity for the remainder of the synthesis, as well as the use of a palladium-mediated direct pyridine functionalization reaction to forge the tetracyclic core.


Weekly chemotherapy with cisplatin and paclitaxel in inoperable non-small cell lung cancer: an extended phase II study.
A phase II study of a cisplatin/paclitaxel combination given on a weekly schedule in the front-line treatment of non-small cell lung cancer (NSCLC) is reported. Treatment consisted of an intravenous infusion of cisplatin, 25 mg/m2, and paclitaxel, 80 mg/m2, every week. Chemotherapy was continued until completion of a 22-week treatment plan, disease progression, persistent toxicity, or patient refusal. Seventy-nine patients entered the study. The median number of infusions per patient was 14 (range 0-22). The median dose-intensity was 75% of that projected. Toxicity was generally acceptable, and never life-threatening. Seven complete responses (pathologically documented in 4 patients) and 27partial responses were observed for an overall response rate of 43%. The estimated median survival and median time to progression was 55 (95% CI: 38-71) and 37 weeks (95% CI: 31-44), respectively. In our experience, the weekly combination of cisplatin and paclitaxel is well tolerated, active and associated with remarkably long survivals.


High molecular weight polyethylene nanospheres: synthesis, physical and mechanical properties--second harmonic generation.
The weak second harmonic light generating from carbon nanotubes are detected. The signal intensity closely related to the density of pi-bonds attributed to the defects in the rolled graphene sheets, which is stimulated to have anharmonic oscillation as strongly affected by the environment. The intensities of SHG are diminished in order of well-aligned multi-wall carbon nanotubes (MWCNTs), randomly-aligned MWCNTs, and then to single-wall CNTs.


The structure of the active site H-cluster of [FeFe] hydrogenase from the green alga Chlamydomonas reinhardtii studied by X-ray absorption spectroscopy.
The [FeFe] hydrogenase (CrHydA1) of the green alga Chlamydomonas reinhardtii is the smallest hydrogenase known and can be considered as a "minimal unit" for biological H(2) production. Due to the absence of additional FeS clusters as found in bacterial [FeFe] hydrogenases, it was possible to specifically study its catalytic iron-sulfur cluster (H-cluster) by X-ray absorption spectroscopy (XAS) at the Fe K-edge. The XAS analysis revealed that the CrHydA1 H-cluster consists of a [4Fe4S] cluster and a diiron site, 2Fe(H), which both are similar to their crystallographically characterized bacterial counterparts. Determination of the individual Fe-Fe distances in the [4Fe4S] cluster ( approximately 2.7 A) and in the 2Fe(H) unit ( approximately 2.5 A) was achieved. Fe-C( horizontal lineO/N) and Fe-S bond lengths were in good agreement with crystallographic data on bacterial enzymes. The loss of Fe-Fe distances in protein purified under mildly oxidizing conditions indicated partial degradation of the H-cluster. Bond length alterations detected after incubation of CrHydA1 with CO and H(2) were related to structural and oxidation state changes at the catalytic Fe atoms, e.g., to the binding of an exogenous CO at 2Fe(H) in CO-inhibited enzyme. Our XAS studies pave the way for the monitoring of atomic level structural changes at the H-cluster during H(2) catalysis.


Demonstration of the in vitro antiviral properties of bovine lymphokine-activated killer (LAK) cells.
In cattle, cells with functional characteristics similar to those of natural killer (NK) cells are difficult to detect. However, lymphokine-activated killer (LAK) cells can be detected readily after in vitro activation of peripheral blood mononuclear leukocytes (PBML) with interleukin-2 (IL-2). In the present study, we demonstrated that IL-2-activated PBML preferentially lyse bovine herpesvirus type 1 (BHV-1)-infected cells and that the cell responsible for the lysis copurified with the cell responsible for lysis of K562. The IL-2-activated effector cells were capable of significant reducing virus production. The reduction in virus yield seemed to be by an interferon (IFN)-independent mechanism, as the amount of IFN induced in effector cells by BHV-1 was not altered by the addition of IL-2. Furthermore, enrichment of cytotoxic cells by passage through nylon wool columns removed the capability of PBML to produce IFN in response to the virus. These results suggest that activation of LAK mechanisms in cattle plays a role in controlling virus spread.


Advanced research on dopamine signaling to develop drugs for the treatment of mental disorders: biochemical and behavioral profiles of phosphodiesterase inhibition in dopaminergic neurotransmission.
Dopamine plays a central role in the regulation of psychomotor functions. The effect of dopamine is largely mediated through the cAMP/PKA signaling cascade and therefore controlled by phosphodiesterases (PDEs). Multiple PDEs with different substrate specificities and subcellular localization are expressed in the striatum, and the functional roles of PDE10A, PDE4, and PDE1B are extensively studied. Biochemical and behavioral profiles of PDE inhibition by selective inhibitors and/or genetic deletion related to dopaminergic neurotransmission are compared among those PDEs. The inhibition of PDE up-regulates cAMP/PKA signaling in three neuronal subtypes, resulting in the stimulation of dopamine synthesis at dopaminergic terminals, the inhibition of dopamine D(2)-receptor signaling in striatopallidal neurons, and the stimulation of dopamine D(1)-receptor signaling in striatonigral neurons. Predominant roles of PDE families or isoforms are implicated in each neuronal subtype: PDE4 at dopaminergic terminals, PDE10A and PDE4 in striatopallidal neurons, and PDE1B in striatonigral neurons. PDE10A and PDE4 inhibition may exhibit D(2) antagonist-like, antipsychotic effects, whereas PDE1B inhibition may exhibit D(1) agonist-like effects in the striatum. Development of PDE isoform-specific inhibitors is essential for better understanding of the function of each PDE isoform and treatment of neuropsychiatric disorders.


The Diagnostic Value of sTWEAK in Acute Ischemic Stroke
Considering the critical role of early diagnosis and management of acute ischemic stroke, biomarkers that can reliable assist in the diagnosis are still needed. These biomarkers should rapidly analyze, have high specificity for brain damage, and be available in the emergency settings for early diagnosis and exclusion of other conditions that mimic acute ischemic stroke. Soluble tumor necrosis factor-like weak inducer of apoptosis, a protein involved in the regulation of several biological functions, could be a potential acute ischemic stroke biomarker. To investigate the diagnostic value of soluble tumor necrosis factor-like weak inducer of apoptosis in patients with acute ischemic stroke and examine the relationship between ischemic area volume determined at diffusion-weighted magnetic resonance imaging and soluble tumor necrosis factor-like weak inducer of apoptosis. A prospective, case-control study. This case-control prospective study included 36 patients with acute ischemic stroke and 36 healthy volunteers. Information on age, sex, presence of chronic disease, neurological examination findings, times of presentation to the emergency department after acute ischemic stroke, soluble tumor necrosis factor-like weak inducer of apoptosis levels, ischemic area volumes at diffusion-weighted magnetic resonance imaging, and 6-month mortality rates after stroke were recorded. The results were analyzed on SPSS 22.0 software (SPSS Inc., Chicago, IL, USA), and p<0.05 was considered statistically significant. A soluble tumor necrosis factor-like weak inducer of apoptosis cut-off value of 995.5 pg/mL exhibited a sensitivity of 80.5% and a positive predictive value of 82.5% with an area under the curve of 0.84 (95% confidence interval: 0.74-0.94; p<0.001). The mean soluble tumor necrosis factor-like weak inducer of apoptosis levels in the acute ischemic stroke group (1968.08±1441.99 μg/L) were significantly higher than those in the control group (704.81±291.72 μg/L) (p<0.001). No correlation was observed between soluble tumor necrosis factor-like weak inducer of apoptosis levels and ischemic area volume measured at diffusion-weighted magnetic resonance imaging (r=-0.008; p=0.07). The mean ischemic area volume was 505.68±381.10 and 60.96±80.89 mm3 in the nonsurviving and surviving patients, respectively (p=0.002). Soluble tumor necrosis factor-like weak inducer of apoptosis can be used in the diagnosis of acute ischemic stroke. However, it is inconclusive in estimating ischemic area volume and early mortality following acute ischemic stroke. Ischemic area volume measured at diffusion-weighted magnetic resonance imaging is a marker of poor prognosis and can be used in predicting early mortality.


Arabinogalactan-Proteins of the Female Sexual Tissue of Nicotiana alata: I. Changes during Flower Development and Pollination.
Arabinogalactan-proteins (AGPs), isolated from the pistils of Nicotiana alata, an ornamental tobacco, are developmentally regulated. Both the total amount and concentration of AGP in the stigma increase during flower development, reaching 10 micrograms AGP/stigma at maturity. In contrast, AGP concentration in the style remains constant throughout the maturation period reaching 12 micrograms AGP/style at maturity. The classes of AGP present in the stigma and style during flower development, separated according to their charge by crossed-electrophoresis, are different and change during development. Pollination of flowers of N. alata with compatible or incompatible pollen results in a significant and reproducible increase in the amount of AGPs in the stigma, but not the style, compared with control unpollinated pistils. Pollination with ethanol vapor inactivated pollen also results in an increase in the amount of AGP in the stigma, but this is less than half that observed following pollination with viable pollen. There are no significant differences in the classes of AGP, based on crossed-electrophoresis, present in the pistil following pollination.


Poly(A) tail affects equilibrium and thermodynamic behavior of tobacco etch virus mRNA with translation initiation factors eIF4F, eIF4B and PABP.
We have investigated the effects of poly(A)-tail on binding of eIF4F, eIF4B and PABP with tobacco etch virus (TEV) IRES RNA. The fluorescence anisotropy data showed that the addition of poly(A)(20) increases the binding affinity of eIF4F·4B and eIF4F·PABP complexes to IRES RNA ~2- and 4-fold, respectively. However, the binding affinity of eIF4F with PK1 was enhanced ~11-fold with the addition of PABP, eIF4B, and poly(A)(20) together. Whereas, poly(A)(20) alone increases the binding affinity of eIF4F·4B·PABP with PK1 RNA about 3-fold, showing an additive effect rather than the large increase in affinity as shown for cap binding. Thermodynamic data showed that PK1 RNA binding to protein complexes in the presence of poly(A)(20) was enthalpy-driven and entropy-favorable. Poly(A)(20) decreased the entropic contribution 75% for binding of PK1 RNA to eIF4F·4B·PABP as compared to eIF4F alone, suggesting reduced hydrophobic interactions for complex formation and an overall conformational change. Overall, these results demonstrate the first direct effect of poly(A) on the equilibrium and thermodynamics of eIF4F and eIF4F·4B·PABP with IRES-RNA.


Natural variation in biogenesis efficiency of individual Arabidopsis thaliana microRNAs.
Like protein-coding genes, loci that produce microRNAs (miRNAs) are generally considered to be under purifying selection, consistent with miRNA polymorphisms being able to cause disease. Nevertheless, it has been hypothesized that variation in miRNA genes may contribute to phenotypic diversity. Here we demonstrate that a naturally occurring polymorphism in the MIR164A gene affects leaf shape and shoot architecture in Arabidopsis thaliana, with the effects being modified by additional loci in the genome. A single base pair substitution in the miRNA complementary sequence alters the predicted stability of the miRNA:miRNA(∗) duplex. It thereby greatly reduces miRNA accumulation, probably because it interferes with precursor processing. We demonstrate that this is not a rare exception and that natural strains of Arabidopsis thaliana harbor dozens of similar polymorphisms that affect processing of a wide range of miRNA precursors. Our results suggest that natural variation in miRNA biogenesis resulting from cis mutations is a common contributor to phenotypic variation in plants.


Women who doctor shop for prescription drugs.
Doctor shopping is a term used to describe a form of diversion of prescription drugs when patients visit numerous prescribers to obtain controlled drugs for illicit use. Gender differences exist in regard to prescription drug abuse and methods of diversion. The purpose of this phenomenological study guided by the existential philosophy of Merleau-Ponty was to understand the lived experience of female doctor shoppers. Interviews were conducted with 14 women, which were recorded, transcribed, and analyzed. Included in the findings are figural aspects of the participants' experience of doctor shopping related to the existential grounds of world, time, body, and others. Four themes emerged from the data: (a) feeding the addiction, (b) networking with addicts, (c) playing the system, and (d) baiting the doctors. The findings suggest several measures that nurses can take to reduce the incidence of doctor shopping and to provide better care for female doctor shoppers.


Sublineage structure analysis of Mycobacterium tuberculosis complex strains using multiple-biomarker tensors.
Strains of Mycobacterium tuberculosis complex (MTBC) can be classified into major lineages based on their genotype. Further subdivision of major lineages into sublineages requires multiple biomarkers along with methods to combine and analyze multiple sources of information in one unsupervised learning model. Typically, spacer oligonucleotide type (spoligotype) and mycobacterial interspersed repetitive units (MIRU) are used for TB genotyping and surveillance. Here, we examine the sublineage structure of MTBC strains with multiple biomarkers simultaneously, by employing a tensor clustering framework (TCF) on multiple-biomarker tensors. Simultaneous analysis of the spoligotype and MIRU type of strains using TCF on multiple-biomarker tensors leads to coherent sublineages of major lineages with clear and distinctive spoligotype and MIRU signatures. Comparison of tensor sublineages with SpolDB4 families either supports tensor sublineages, or suggests subdivision or merging of SpolDB4 families. High prediction accuracy of major lineage classification with supervised tensor learning on multiple-biomarker tensors validates our unsupervised analysis of sublineages on multiple-biomarker tensors. TCF on multiple-biomarker tensors achieves simultaneous analysis of multiple biomarkers and suggest a new putative sublineage structure for each major lineage. Analysis of multiple-biomarker tensors gives insight into the sublineage structure of MTBC at the genomic level.


Removable colored coatings based on calcium alginate hydrogels.
This article describes the creation of a nontoxic, biodegradable coating using calcium alginate and FD&C approved dyes. The coating is robust but is rapidly removed upon treatment with disodium ethylenediamine tetraacetate (EDTA). Dye leaching from calcium alginate films was studied, and it was determined that the efficiency of dye retention is proportional to the degree of cross-linking. Degradation rates were studied on calcium alginate beads serving as a model for a coating. We determined that degradation rates depend on the gel's cross-linking and on the amount of EDTA used. Bead size also influenced the degradation rates; smaller beads degraded faster than larger beads. We show that the coating can be used as an easily removable and environmentally friendly logotype on an artificial turf surface. Applications of these coatings can be extended to food, cosmetic, medicinal, and textile uses and to wherever nontoxic, easily removable colored coating is desired.


[Determination of trace lead in water and milk tea powder samples with organic coprecipitation-flame atomic absorption spectrometric].
A method was proposed for the determination of trace lead with flame atomic absorption spectrometry after preconcentration of lead by rapid coprecipitation technique with PAR-Fe (III) at pH 6.0. The analytical parameters including pH, amount of iron (III), amount of reagent, the standing time of the precipitate, etc., were examined. The detection limits (DL) were found to be 18.7 microg x L(-1) for Pb (II). In analysis of lake water and the milk tea powder samples, RSD's and the standard addition recovery of this method were in the ranges of 1.03%-2.24% and 94.2%-98.3% respectively. The effect of matrix can be overcome by the method and the results are satisfyiog. The method shows good application prospect in the determination of trace lead owing to its rapidness and reproducibility.


Expression of rat cathepsin S in phagocytic cells.
Cysteine lysosomal proteases are essential for turnover of intracellular and extracellular proteins. These enzymes are strongly implicated in normal and pathological processes involving tissue remodeling. Among the cysteine proteases, cathepsin S seems to be best suited for such a process since it retains most of its enzymatic activity at neutral pH. In situ hybridization analyses of the adult rat brain, spleen, and lung reveal that cathepsin S mRNA is preferentially expressed in cells of mononuclear-phagocytic origin. After entorhinal cortex lesion of adult rat brain (a paradigm for neuronal degeneration and reactive synaptogenesis), cathepsin S mRNA is dramatically increased in activated microglia in the deafferented dentate gyrus and in macrophages at the wound site, suggesting a role in lesion-induced tissue remodeling. This possibility is further supported by the finding that cathepsin S degrades a number of extracellular matrix molecules at neutral pH and by the finding that inflammatory mediators stimulate its secretion from the microglia and macrophages. These data suggest that cathepsin S is an important player in degenerative disorders associated with the cells of the mononuclear phagocytic system.


Patient selection for Port-Access multi vessel revascularization.
Minimally invasive multivessel revascularization is rarely performed due to the difficult exposure of the aorta as well as the complete coronary anatomy through a small thoracotomy. The Port-Access technique bears additional contraindications for this procedure, which limits its potential as compared with other approaches to 'less invasive surgery'. Our aim was to show the applicability of this surgical technique to a wide range of patients with coronary artery disease. In our initial experience with this method (31 patients), the quality of anastomoses, graft patency, and clinical outcome are good, and do not differ from standard multivessel coronary artery bypass grafting. Port-Access multivessel revascularization can be performed safely and is appropriate for a large patient population.


SMS 201-995, an octapeptide somatostatin analogue. Assignment of the 1H 500 MHZ n.m.r. spectra and conformational analysis of SMS 201-995 in dimethylsulfoxide.
The possible conformations of SMS 201-995, an active analogue of somastostatin, have been studied in dimethylsulfoxide solution by 500 MHz proton n.m.r. spectroscopy. The assignments have been made by use of 2D-correlated methods to detect long-range coupling connectivities in aromatic residues and between the alpha protons of consecutive residues. NOESY experiments enabled us to correlate amide and alpha protons of neighbouring amino acid residues, which indicate a less flexible situation than in water. Measurements of temperature coefficients of the amide protons, of NH-C alpha H coupling constants and NOE effects are in favour of one predominant conformation with a beta turn, of type II', involving amino acids Phe3 to Thr6.


IL-17-Dependent Dysregulated Cutaneous Immune Homeostasis in the Absence of the Wiskott-Aldrich Syndrome Protein.
Wiskott-Aldrich Syndrome (WAS) is characterized by recurrent infections, thrombocytopenia, and eczema. Here, we show that WASp-deficient mice on a BALB/c background have dysregulated cutaneous immune homeostasis with increased leukocyte accumulation in the skin, 1 week after birth. Increased cutaneous inflammation was associated with epithelial abnormalities, namely, altered keratinization, abnormal epidermal tight junctional morphology and increased trans-epidermal water loss; consistent with epidermal barrier dysfunction. Immune and physical barrier disruption was accompanied by progressive skin dysbiosis, highlighting the functional significance of the disrupted cutaneous homeostasis. Interestingly, the dysregulated immunity in the skin preceded the systemic elevation in IgE and lymphocytic infiltration of the colonic lamina propria associated with WASp deficiency. Mechanistically, the enhanced immune cell accumulation in the skin was lymphocyte dependent. Elevated levels of both Type 2 (IL-4, IL-5) and Type 17 (IL-17, IL-22, IL-23) cytokines were present in the skin, as well as the 'itch' factor IL-31. Unexpectedly, the canonical WAS-associated cytokine IL-4 did not play a role in the immune dysfunction. Instead, IL-17 was critical for skin immune infiltration and elevation of both Type 2 and Type 17 cytokines. Our findings reveal a previously unrecognized IL-17-dependent breakdown in immune homeostasis and cutaneous barrier integrity in the absence of WASp, targeting of which may provide new therapeutic possibilities for the treatment of skin pathologies in WAS patients.