Trichosanthes kirilowii Extract Promotes Wound Healing through the Phosphorylation of ERK1/2 in Keratinocytes.
The proliferation of keratinocytes is one of the important steps in the wound-healing process, which is regulated by various signals. Prior studies have shown that Trichosanthes kirilowii extract has the ability to promote angiogenesis. Therefore, in this study, we tested the wound-healing efficacy of Trichosanthes kirilowii extract with respect to promoting keratinocyte proliferation. A total of 100 μg/mL of Trichosanthes kirilowii extract treatment improved 145.38% of keratinocyte proliferation compared with DMSO-treated control in an MTT assay and increased 238.2% of wound closure by re-epithelialization in an in vitro wound-healing assay. Trichosanthes kirilowii extract promoted ERK1/2 phosphorylation in western blot analysis and induced the expression of the c-fos and c-jun (AP-1 transcription factors), cyclins (cell cycle regulator), and growth factors CTGF and VEGF (stimulator of angiogenesis) in qRT-PCR analysis. An in vivo wound-healing assay showed that Trichosanthes kirilowii extract improved wound healing, and the significant difference in wound closure compared with DMSO-treated control was shown on days 6 and 7 with a mouse model. Taken together, we demonstrate that Trichosanthes kirilowii extract promotes the proliferation of keratinocytes by activating ERK1/2 and increasing the mRNA expression of c-fos, c-jun, CTGF, and VEGF. Therefore, we suggest Trichosanthes kirilowii extract as a new component for skin care and as a wound-healing substance.


Assessment of testicular biopsy after cessation of maintenance chemotherapy in childhood acute lymphoblastic leukemia: a report from the Children's Cancer and Leukemia Study Group.
Among 484 male patients with acute lymphoblastic leukemia (ALL) registered into the protocols CCLSG 811, 841 and 874, from 1981 through 1990, 246 patients completed their protocols and were in continuous complete remission (CCR) for more than 3 years. One hundred and seven patients received bilateral testicular biopsies at the time of cessation of maintenance chemotherapy. Eight patients (7.5%) were found to have occult testicular leukemia (TL). Three of them did not receive any additional therapy and all suffered subsequent relapses; one bone marrow relapse and two testicular relapses. The other 3 patients received testicular radiation combined with an additional 2 years of chemotherapy, resulting in CCR for more than 6 years 10 months, 7 years 6 months, and 8 years 6 months. One with chemotherapy alone and another with radiation alone showed subsequent relapse. Overt TL after negative initial biopsy was developed in 3 (3.0%) of the 99 patients. All of them received testicular radiation with chemotherapy, resulting again in CCR for more than 1 year 0 months and 5 years 3 months; one patient showed relapse in testes and bone marrow after 3 years 8 months of CCR. These studies suggested that occult TL has an adverse prognostic significance unless retrieval chemotherapy is given and that performance of testicular biopsy at completion of maintenance chemotherapy is not contributory to prolongation of disease-free survival for males with ALL because the treatment employing testicular radiation plus retrieval chemotherapy for both occult TL and isolated overt TL after off-therapy is similarly very effective.


Clinical guidelines and self-assessment in dental education.
This paper describes an educational programme where clinical guidelines are used in dental education and assesses the value of these guidelines as perceived by undergraduate students. It presents a comparison of students' own assessments of their performance with those of their instructors, according to such guidelines. The educational programme at the Faculty of Odontology, Lund University, is described through 3 main principles: problem-based learning, adoption of a holistic attitude to patient care and the promotion of oral health. The dental curriculum occupies 5 years comprising 10 semesters. 41, 5th semester students (16 male and 25 female) participated in the study. Students were introduced to guidelines in group discussions at the beginning of the semester. At the completion of a patient's planned course of treatment, the outcome was formally assessed both by students and clinical instructors. Performance was scored in various categories as "excellent", "acceptable" or "unacceptable" according to the standards defined. A total of 1373 scores were made by both students and instructors. 88% cent of students used the guidelines often or almost always and found them useful. Overall, no students received an "unacceptable" score and 40% of students achieved a score of "excellent". Female students tended towards a higher score, but this was not significant. Instructors' and students' scores agreed in almost 90% of instances. Students under-scored their performance more frequently than they over-scored it. It appears that the use of clinical guidelines may encourage an increased awareness of the decision-making processes involved in clinical practice, but it must be acknowledged that these guidelines are complex both in derivation and application.


Safety assessment of the Clostridium butyricum MIYAIRI 588® probiotic strain including evaluation of antimicrobial sensitivity and presence of Clostridium toxin genes in vitro and teratogenicity in vivo.
Probiotics are live microorganisms ingested for the purpose of conferring a health benefit on the host. Development of new probiotics includes the need for safety evaluations that should consider factors such as pathogenicity, infectivity, virulence factors, toxicity, and metabolic activity. Clostridium butyricum MIYAIRI 588(®) (CBM 588(®)), an anaerobic spore-forming bacterium, has been developed as a probiotic for use by humans and food animals. Safety studies of this probiotic strain have been conducted and include assessment of antimicrobial sensitivity, documentation of the lack of Clostridium toxin genes, and evaluation of CBM 588(®) on reproductive and developmental toxicity in a rodent model. With the exception of aminoglycosides, to which anaerobes are intrinsically resistant, CBM 588(®) showed sensitivity to all antibiotic classes important in human and animal therapeutics. In addition, analysis of the CBM 588(®) genome established the absence of genes for encoding for α, β, or ε toxins and botulin neurotoxins types A, B, E, or F. There were no deleterious reproductive and developmental effects observed in mice associated with the administration of CBM 588(®) These data provide further support for the safety of CBM 588(®) for use as a probiotic in animals and humans.


Differential Effect of Endurance Training on Mitochondrial Protein Damage, Degradation, and Acetylation in the Context of Aging.
Acute aerobic exercise increases reactive oxygen species and could potentially damage proteins, but exercise training (ET) enhances mitochondrial respiration irrespective of age. Here, we report a differential impact of ET on protein quality in young and older participants. Using mass spectrometry we measured oxidative damage to skeletal muscle proteins before and after 8 weeks of ET and find that young but not older participants reduced oxidative damage to both total skeletal muscle and mitochondrial proteins. Young participants showed higher total and mitochondrial derived semitryptic peptides and 26S proteasome activity indicating increased protein degradation. ET however, increased the activity of the endogenous antioxidants in older participants. ET also increased skeletal muscle content of the mitochondrial deacetylase SIRT3 in both groups. A reduction in the acetylation of isocitrate dehydrogenase 2 was observed following ET that may counteract the effect of acute oxidative stress. In conclusion aging is associated with an inability to improve skeletal muscle and mitochondrial protein quality in response to ET by increasing degradation of damaged proteins. ET does however increase muscle and mitochondrial antioxidant capacity in older individuals, which provides increased buffering from the acute oxidative effects of exercise.


Intake of free sugars and micronutrient dilution in Australian adults.
The negative health effect of excessive intake of free sugars has been gaining increasing public awareness. This secondary analysis aimed to evaluate the impact of free-sugar intake on micronutrient dilution, and estimate a threshold level of free-sugar intake at which a decrease in micronutrient intake becomes evident, based on data from the Australian Health Survey 2011-2012. Dietary data from adult respondents (weighted n = 6150) who had completed two 24-h recalls were analyzed. A published 10-step methodology was adopted and used to estimate the free-sugar intake of the respondents. Six modified cut-offs for percentage of energy of free sugars (%EFS) were created based on recommendations from the WHO and the Institute of Medicine to examine the association between %EFS on micronutrient intakes. Estimated marginal means and SEs were calculated using ANCOVA. Logistic regression was used to calculate the ORs of not meeting the nutrient reference values for Australia and New Zealand for each micronutrient with an increase in free-sugar intake. Analyses were adjusted for age, sex, socioeconomic status, country of birth, whether dieting, smoking status, and remoteness of living area. Peak intake for most micronutrients was observed at %EFS between 5% and <15%. A significant reduction in most micronutrient intakes was observed at >25%EFS. At <5%EFS, some micronutrient intakes were reduced. Only small variations in micronutrient consumptions were observed when %EFS was between 5% and 25%. Core food intake decreased and discretionary food increased with an increase in free-sugar intake. A high free-sugar intake, particularly >25%EFS, was found to have a significant diluting effect on most nutrients. However, a free-sugar intake <5%EFS may increase the risk of undesirably low micronutrient consumption related to inadequate total energy intake. This secondary analysis was registered at anzctr.org.au as ACTRN12617000917336.


Validity and reliability of the DANU sports system for walking and running gait assessment.
Objective. Gait assessments have traditionally been analysed in laboratory settings, but this may not reflect natural gait. Wearable technology may offer an alternative due to its versatility. The purpose of the study was to establish the validity and reliability of temporal gait outcomes calculated by the DANU sports system, against a 3D motion capture reference system.Approach. Forty-one healthy adults (26 M, 15 F, age 36.4 ± 11.8 years) completed a series of overground walking and jogging trials and 60 s treadmill walking and running trials at various speeds (8-14 km hr-1), participants returned for a second testing session to repeat the same testing.Main results. For validity, 1406 steps and 613 trials during overground and across all treadmill trials were analysed respectively. Temporal outcomes generated by the DANU sports system included ground contact time, swing time and stride time all demonstrated excellent agreement compared to the laboratory reference (intraclass correlation coefficient (ICC) > 0.900), aside from ground contact time during overground jogging which had good agreement (ICC = 0.778). For reliability, 666 overground and 511 treadmill trials across all speeds were examined. Test re-test agreement was excellent for all outcomes across treadmill trials (ICC > 0.900), except for swing time during treadmill walking which had good agreement (ICC = 0.886). Overground trials demonstrated moderate to good test re-test agreement (ICC = 0.672-0.750), which may be due to inherent variability of self-selected (rather than treadmill set) pacing between sessions.Significance. Overall, this study showed that temporal gait outcomes from the DANU Sports System had good to excellent validity and moderate to excellent reliability in healthy adults compared to an established laboratory reference.


Optimizing Wound Bed Preparation With Collagenase Enzymatic Debridement.
Difficult-to-heal and chronic wounds affect tens of millions of people worldwide. In the U.S. alone, the direct cost for their treatment exceeds $25 billion. Yet despite advances in wound research and treatment that have markedly improved patient care, wound healing is often delayed for weeks or months. For venous and diabetic ulcers, complete wound closure is achieved in as few as 25%-50% of chronic or hard-to-heal wounds. Wound bed preparation and the consistent application of appropriate and effective debridement techniques are recommended for the optimized treatment of chronic wounds. The TIME paradigm (Tissue, Inflammation/infection, Moisture balance and Edge of wound) provides a model to remove barriers to healing and optimize the healing process. While we often think of debridement as an episodic event that occurs in specific care giver/patient interface. There is the possibility of a maintenance debridement in which the chronic application of a medication can assist in both the macroscopic and microscopic debridement of a wound. We review the various debridement therapies available to clinicians in the United States, and explore the characteristics and capabilities of clostridial collagenase ointment (CCO), a type of enzymatic debridement, that potentially allows for epithelialization while debriding. It appears that in the case of CCO it may exert this influences by removal of the necrotic plug while promoting granulation and sustaining epithelialization. It is also easily combined with other methods of debridement, is selective to necrotic tissue, and has been safely used in various populations. We review the body of evidence has indicated that this concept of maintenance debridement, especially when combined episodic debridement may add a cost an efficacious, safe and cost-effective choice for debridement of cutaneous ulcers and burn wounds and it will likely play an expanding role in all phases of wound bed preparation.


Prognosis of Primary Liver Cancer Based on LI-RADS Classification with Extracellular Agent-Enhanced MRI.
The prognostic value of the Liver Imaging Reporting and Data System (LI-RADS) 2018 in differentiating hepatocellular carcinoma (HCC) from other primary liver cancers (PLC) with cirrhosis is unclear. We aim to evaluate the value of LI-RADS 2018 with agent-enhanced MRI in the postoperative prognosis of PLC patients with cirrhosis. Between 2016 and 2021, 432 patients with cirrhosis and surgically proven single primary liver cancer were retrospectively evaluated. Two radiologists evaluated the preoperative MRI features independently and assigned each lesion a LI-RADS category. Overall survival (OS), recurrence-free survival (RFS), and their associated factors were evaluated by using the Kaplan-Meier method, Log rank test, and Cox proportional hazards model. The mean age of 432 patients (239 HCCs, 93 ICCs, and 100 cHCC-CCAs) was 57.27±10.92 years. The LR-M category showed poorer OS and RFS than the LR-4 or LR-5 category did for all primary liver cancers (P <0.001 for both), and so did HCCs with tumor size less than 30mm (P =0.003 and P =0.04, respectively). In the multivariable analysis, the LI-RADS category and tumor size > 30 mm had independent correlations with OS and RFS (all P < 0.05). Multivariable Cox analysis identified rim arterial phase hyperenhancement (APHE) as independent determinants of poor OS and RFS in primary liver cancers (all P < 0.05). The LI-RADS categories can predict the postsurgical prognosis of primary liver cancers independently.


Thiadiazole and Thiazole Derivatives as Potential Antimicrobial Agents.
This review summarizes data on heterocyclic systems with thiadiazole and thiazole fragments in molecules as promising antimicrobial agents. Thiadiazole and thiazole backbones are the most favored and well-known heterocycles, a common and essential feature of various drugs. These scaffolds occupy a central position and are the main structural components of numerous drugs with a wide spectrum of action. These include antimicrobial, antituberculous, anti-inflammatory, analgesic, antiepileptic, antiviral, and anticancer agents. The research is based on bibliosemantic and analytical methods using bibliographic and abstract databases, as well as databases of chemical compounds. This review reports on thiadiazole and thiazole derivatives, which have important pharmacological properties. We are reviewing the structural modifications of various thiadiazole and thiazole derivatives, more specifically, the antimicrobial activity reported over the last years, as we have taken this as our main research area. 80 compounds were illustrated, and various derivatives containing hydrazone bridged thiazole and pyrrole rings, 2-pyridine and 4-pyridine substituted thiazole derivatives, compounds containing di-, tri- and tetrathiazole moieties, Spiro-substituted 4-thiazolidinone-imidazoline-pyridines were analyzed. Derivatives of 5-heteroarylidene-2,4-thiazolidinediones, fluoroquinolone-thiadiazole hybrids, and others. 1,3,4-thiadiazoles and thiazoles are valuable resource for researchers engaged in rational drug design and development in this area.


Antiproliferative effect of the jararhagin toxin on B16F10 murine melanoma.
Malignant melanoma is a less common but highly dangerous form of skin cancer; it starts in the melanocytes cells found in the outer layer of the skin. Jararhagin toxin, a metalloproteinase isolated from Bothrops jararaca snake venom acts upon several biological processes, as inflammation, pain, platelet aggregation, proliferation and apoptosis, though not yet approved for use, may one day be employed to treat tumors. B16F10 murine melanoma cells were treated with jararhagin (jara), a disintegrin-like metalloproteinase isolated from Bothrops jararaca snake venom, and jari (catalytic domain inactivated with 1,10-phenanthroline). Viability and adhesion cells were evaluated by MTT assay. The expression of caspase-3 active, phases of the cell cycle and apoptosis were assessed by flow cytometry. We analyze in vivo the effects of jararhagin on melanoma growth, apoptosis and metastasis. The tumor cells acquired round shapes, lost cytoplasmic expansions, formed clusters in suspension and decreased viability. Jari was almost 20 times more potent toxin than jara based on IC50 values and on morphological changes of the cells, also observed by scanning electron microscopy. Flow cytometry analysis showed 48.3% decrease in the proliferation rate of cells and 47.2% increase in apoptosis (jara) and necrosis (jari), following 1.2 μM jara and 0.1 μM jari treatments. Caspase-3 activity was increased whereas G0/G1 cell cycle phase was on the decline. Proliferative rate was assessed by staining with 5,6-carboxyfluoresceindiacetate succinimidyl ester, showing a significant decrease in proliferation at all concentrations of both toxins. In vivo treatment of the toxins was observed reduction in the incidence of nodules, and metastasis and antiproliferative inhibition capacity. This data strengthens the potential use jararhagin as an anti-neoplastic drug.


Hydrated Ion Clusters in Hydrophobic Liquid: Equilibrium Distribution, Kinetics, and Implications.
Hydrophilic ions in oil phase tend to be hydrated in the presence of trace water and form hydrated clusters. The present paper elucidates the equilibrium size distribution of hydrated ion clusters and the microscopic rates of adsorption and desorption of water with the help of molecular dynamics simulations. The size distribution is derived from reversible work of hydration, which is nearly constant over the hydration number except for small clusters. The intrinsic rate constants of adsorption and desorption are evaluated to be in several psec order after correcting the diffusion. The microscopic hydration properties of ions in the oil phase play key roles in chemical reactions involving both hydrophilic and hydrophobic reactants as well as in the transport and reactivity of the ions in oil phase and at the water-oil interface.


[Relationship between population dynamic of freshwater leech Whitmania laevis and chemical factors of water].
The population size of freshwater leech Whitmania laevis had been observed every month in Guangzhou kaleyard slot from 1994 to 1998, and ten chemical factors of water were measured. The relationship between the population dynamic of W. laevis and chemical factors was studied by stepwise regression analysis method. The results showed that the population size dynamic of W. laevis was different every year. The maximum of population size appeared in April, May, and June, and the total population size of three months was accounted for 57.14-71.18% of whole year. The major chemical factors that effected on the annual population size were different every year, and the factors included pH value, PO(4)3-, Na+, Ca2+, and Mg2+. Fe3+ was the chief chemical factor effecting on population size during the 5 years observation. The behavior of leech W. laevis was also described in this paper.


Development and validation of a UHPLC-MS/MS method for quantification of the prodrug remdesivir and its metabolite GS-441524: a tool for clinical pharmacokinetics of SARS-CoV-2/COVID-19 and Ebola virus disease.
Remdesivir has received significant attention for its potential application in the treatment of COVID-19, caused by SARS-CoV-2. Remdesivir has already been tested for Ebola virus disease treatment and found to have activity against SARS and MERS coronaviruses. The remdesivir core contains GS-441524, which interferes with RNA-dependent RNA polymerases alone. In non-human primates, following IV administration, remdesivir is rapidly distributed into PBMCs and converted within 2 h to the active nucleoside triphosphate form, while GS-441524 is detectable in plasma for up to 24 h. Nevertheless, remdesivir pharmacokinetics and pharmacodynamics in humans are still unexplored, highlighting the need for a precise analytical method for remdesivir and GS-441524 quantification. The validation of a reliable UHPLC-MS/MS method for remdesivir and GS-441524 quantification in human plasma. Remdesivir and GS-441524 standards and quality controls were prepared in plasma from healthy donors. Sample preparation consisted of protein precipitation, followed by dilution and injection into the QSight 220 UHPLC-MS/MS system. Chromatographic separation was obtained through an Acquity HSS T3 1.8 μm, 2.1 × 50 mm column, with a gradient of water and acetonitrile with 0.05% formic acid. The method was validated using EMA and FDA guidelines. Analyte stability has been evaluated and described in detail. The method successfully fulfilled the validation process and it was demonstrated that, when possible, sample thermal inactivation could be a good choice in order to improve biosafety. This method represents a useful tool for studying remdesivir and GS-441524 clinical pharmacokinetics, particularly during the current COVID-19 outbreak.


Estimating feedforward vs. feedback control of speech production through kinematic analyses of unperturbed articulatory movements.
To estimate the contributions of feedforward vs. feedback control systems in speech articulation, we analyzed the correspondence between initial and final kinematics in unperturbed tongue and jaw movements for consonant-vowel (CV) and vowel-consonant (VC) syllables. If movement extents and endpoints are highly predictable from early kinematic information, then the movements were most likely completed without substantial online corrections (feedforward control); if the correspondence between early kinematics and final amplitude or position is low, online adjustments may have altered the planned trajectory (feedback control) (Messier and Kalaska, 1999). Five adult speakers produced CV and VC syllables with high, mid, or low vowels while movements of the tongue and jaw were tracked electromagnetically. The correspondence between the kinematic parameters peak acceleration or peak velocity and movement extent as well as between the articulators' spatial coordinates at those kinematic landmarks and movement endpoint was examined both for movements across different target distances (i.e., across vowel height) and within target distances (i.e., within vowel height). Taken together, results suggest that jaw and tongue movements for these CV and VC syllables are mostly under feedforward control but with feedback-based contributions. One type of feedback-driven compensatory adjustment appears to regulate movement duration based on variation in peak acceleration. Results from a statistical model based on multiple regression are presented to illustrate how the relative strength of these feedback contributions can be estimated.


Impact of zinc supplementation on morbidity from diarrhea and respiratory infections among rural Guatemalan children.
A community-based, randomized, double-blind intervention trial was conducted to measure the impact of zinc supplementation on young Guatemalan children's morbidity from diarrhea and respiratory infections. Children aged 6 to 9 months were randomly assigned to receive 4 mL of a beverage containing 10 mg of zinc (as zinc sulfate) daily (7 d/wk) for 7 months (n = 45) or a placebo (n = 44). Morbidity data were collected daily. Diagnoses of diarrhea, fever, and anorexia were based on mothers' definitions. Respiratory infections were defined as the presence of at least two of the following symptoms: runny nose, cough, wheezing, difficulty breathing, or fever. High rates of diarrhea and respiratory infections were reported. Children from the placebo group had a 20% episodic prevalence of diarrhea, with 8 episodes/100 d, and a 7% episodic prevalence of respiratory infections, with 3 episodes/100 d. The median incidence of diarrhea among children who received zinc supplementation was reduced by 22% (Wilcoxon rank test), with larger reductions among boys and among children with weight-for-length at baseline lower than the median of the sample (39% reductions in both subgroups). Zinc supplementation also produced a 67% reduction in the percentage of children who had one or more episodes of persistent diarrhea (chi2 test). No significant effects were found on the episodic prevalence of diarrhea, the number of days per episode, or the episodic prevalence or incidence of respiratory infections. The large impact of zinc supplementation on diarrhea incidence suggests that young, rural Guatemalan children may be zinc deficient and that zinc supplementation may be an effective intervention to improve their health and growth.


Statins improve survival in patients previously treated with nivolumab for advanced non-small cell lung cancer: An observational study.
There are a number of suggested predictive factors of nivolumab for non-small cell lung cancer (NSCLC), however, there is not enough evidence to determine a single factor that can predict the efficacy of nivolumab. As the progress of biomarkers for cancer treatment is improving, it has been speculated that certain clinical factors serve an important role when predicting the outcome of chemotherapy. A total of 67 patients treated with nivolumab for NSCLC from 2016-2017 were prospectively investigated. Age, sex, the Eastern Cooperative Oncology Group Performance Status, histology, epidermal growth factor receptor (EGFR) mutation, history of chemotherapy, smoking status, use of statins, use of fibrates, use of dipeptidyl peptidase-4 (DPP-4) inhibitors, and use of metformin were examined as clinical factors. Statistical analyses were performed using the Kaplan-Meier method and Cox regression adjusted for risk factors and the tumor response of 67 patients was assessed. The patients had a median age of 67 years (range, 36-87 years), and 46 males and 21 females were enrolled; performance status 0/1 was 59. Cases were categorized as adenocarcinoma (n=41), squamous cell carcinoma (n=17) and other (n=9). A total of 13 patients (19.4%) had EGFR mutations. These clinical factors were not statistically significant in overall survival (OS). Clinical laboratory findings, complications and use of medical agents including antidiabetes mellitus or lipidemia were also analyzed. Statins exhibited statistical significance for response (P=0.02). Time-to-treatment failure (TTF) in statin-use group was not reached [95% confidence interval (CI): 1.9-not reached] and was 4.0 months (95% CI: 2.0-5.4) in the non-statin group (P=0.039). The median OS in statin-use group was not reached (95% CI: 8.7-not reached) and was 16.5 months (95% CI: 7.5-not reached) in the non-statin group (P=0.058). NSCLC patients previously treated with nivolumab who were administered statins exhibited an increased response rate and longer TTF. This response was not statistically significant in OS.


Inhibition of PbGP43 expression may suggest that gp43 is a virulence factor in Paracoccidioides brasiliensis.
Glycoprotein gp43 is an immunodominant diagnostic antigen for paracoccidioidomycosis caused by Paracoccidioides brasiliensis. It is abundantly secreted in isolates such as Pb339. It is structurally related to beta-1,3-exoglucanases, however inactive. Its function in fungal biology is unknown, but it elicits humoral, innate and protective cellular immune responses; it binds to extracellular matrix-associated proteins. In this study we applied an antisense RNA (aRNA) technology and Agrobacterium tumefaciens-mediated transformation to generate mitotically stable PbGP43 mutants (PbGP43 aRNA) derived from wild type Pb339 to study its role in P. brasiliensis biology and during infection. Control PbEV was transformed with empty vector. Growth curve, cell vitality and morphology of PbGP43 aRNA mutants were indistinguishable from those of controls. PbGP43 expression was reduced 80-85% in mutants 1 and 2, as determined by real time PCR, correlating with a massive decrease in gp43 expression. This was shown by immunoblotting of culture supernatants revealed with anti-gp43 mouse monoclonal and rabbit polyclonal antibodies, and also by affinity-ligand assays of extracellular molecules with laminin and fibronectin. In vitro, there was significantly increased TNF-α production and reduced yeast recovery when PbGP43 aRNA1 was exposed to IFN-γ-stimulated macrophages, suggesting reduced binding/uptake and/or increased killing. In vivo, fungal burden in lungs of BALB/c mice infected with silenced mutant was negligible and associated with decreased lung ΙΛ-10 and IL-6. Therefore, our results correlated low gp43 expression with lower pathogenicity in mice, but that will be definitely proven when PbGP43 knockouts become available. This is the first study of gp43 using genetically modified P. brasiliensis.


The effectiveness of aerobic exercise for pain management in patients with fibromyalgia.
The effectiveness of exercise therapy is being increasingly studied in patients with fibromyalgia. The aim of our systematic review was to determine the effectiveness of different types of aerobic exercise programs for pain management in patients with fibromyalgia. The literature search was performed by two independent researchers in the PubMed, CINAHL and PEDro databases using various combinations of the following keywords: fibromyalgia, pain and aerobic exercise. Studies were eligible if they included adults diagnosed with fibromyalgia and examined the effectiveness of at least one aerobic exercise program on pain management. A total of 14 randomized controlled trials were screened in full-text, nine of which were included in the systematic review. Overall, our results indicate that aerobic exercise is effective for pain management in patients with fibromyalgia. The results of the aerobic exercise programs were more effective for pain management than stretching exercises, but did not differ significantly from those of pilates, muscle strengthening exercises, relaxation techniques and stress management treatment. Additional high-quality studies are warranted to determine accurate clinical guidelines in terms of aerobic exercise programs for pain management in patients with fibromyalgia.


Using multidimensional Rasch to enhance measurement precision: initial results from simulation and empirical studies.
This study aimed to explore the effect on measurement precision of multidimensional, as compared with unidimensional, Rasch measurement for constructing measures from multidimensional Likert-type scales. Many educational and psychological tests are multidimensional but common practice is to ignore correlations among the latent traits in these multidimensional scales in the measurement process. These practices may have serious validity and reliability implications. This study made use of both empirical data from 208,083 students, and simulated data simulated by 24 systematic combinations, each replicated 1000 times, of three conditions, namely, sample size, degree of dimensionality, and scale length to compare unidimensional and multidimensional approaches and to identify effects of sample size, dimensionality and scale length on measurement precision. Results showed that the multidimensional Rasch approach yielded more precise estimates than did unidimensional approach if the two dimensions were strongly correlated. The effect was more pronounced for long scales.


JNK regulates autocrine expression of TGF-beta1.
The c-Jun NH2-terminal kinase (JNK) has been implicated in the function of transforming growth factor beta (TGF-beta). To test the role of JNK, we examined the effect of compound disruption of the murine genes that encode the ubiquitously expressed isoforms of JNK (Jnk1 and Jnk2). We report that JNK-deficient fibroblasts isolated from Jnk1-/- Jnk2-/- mice constitutively express TGF-beta1. Complementation studies demonstrate that JNK is a repressor of Tgf-beta1 gene expression. This mechanism of regulation of TGF-beta1 expression by JNK represents an unexpected form of cross-talk between two important signaling pathways. Together, these data demonstrate that the JNK pathway may contribute to the regulation of autocrine TGF-beta1-mediated biological responses in vivo.


Disorder in crystals of dioxofluorotungstates, (NH4)2WO2F4 and Rb2WO2F4.
Dioxotetrafluorotungstates (NH4)2WO2F4 and Rb2WO2F4 [(I) at 297 K and (II) at 133 K] and Rb2WO2F4 (III) were synthesized in a single-crystal form and their structures were determined by X-ray diffraction. Two independent states of the cis-WO2F4 octahedron are characteristic of static and dynamic disorder in structure (I). Dynamically disordered W2 is displaced from the symmetry axis producing four possible orientations of anion that permits O and F atoms to be identified in separate orientations owing to the inherent differences between W-O and W-F bonding. After the phase transition at lower temperature (201 K), (I) transforms into the twin structure (II) with complete O/F ordering. Structure (III) is characterized by full O/F static disorder without any phase transitions at lower temperature.


Regulation of lipolysis in isolated adipocytes of rainbow trout (Oncorhynchus mykiss): the role of insulin and glucagon.
In the present study, we have examined the effects of insulin and glucagon on the lipolysis of rainbow trout (Oncorhynchus mykiss). To this end, adipocytes were isolated from mesenteric fat and incubated in the absence (basal lipolysis) or presence of different concentrations of insulin and glucagon. In addition, to further elucidate the effects of these hormones in vivo on adipocyte lipolysis, both fasting and intraperitoneal glucagon injection experiments were performed. Basal lipolysis, measured as the glycerol released in the adipocyte medium, increased proportionally with cell concentration and incubation time. Cell viability was verified by measuring the release of lactate dehydrogenase (LDH) activity in the medium. Insulin (at doses of 35 and 350 nM) decreased lipolysis in isolated adipocytes of rainbow trout in vitro, while glucagon was clearly lipolytic at concentrations of 10 and 100 nM. Furthermore, hypoinsulinemia induced by fasting, as well as glucagon injection, significantly increased lipolysis in isolated adipocytes approximately 1.5- and 1.4-fold, respectively, when compared with adipocytes from control fish. Our data demonstrate that lipolysis, as measured in isolated adipocytes of rainbow trout, can be regulated by both insulin and glucagon. These results not only indicate that insulin is an important hormone in lipid deposition via its anti-lipolytic effects on rainbow trout adipocytes, but also reveal glucagon as a lipolytic hormone, as shown by both in vitro and in vivo experiments.


Abrogation of Marek's disease virus replication using CRISPR/Cas9.
Marek's disease virus (MDV) is a highly cell-associated alphaherpesvirus that causes deadly lymphomas in chickens. While vaccination protects against clinical symptoms, MDV field strains can still circulate in vaccinated flocks and continuously evolve towards greater virulence. MDV vaccines do not provide sterilizing immunity, allowing the virus to overcome vaccine protection, and has increased the need for more potent vaccines or alternative interventions. In this study, we addressed if the CRISPR/Cas9 system can protect cells from MDV replication. We first screened a number of guide RNAs (gRNAs) targeting essential MDV genes for their ability to prevent virus replication. Single gRNAs significantly inhibited virus replication, but could result in the emergence of escape mutants. Strikingly, combining two or more gRNAs completely abrogated virus replication and no escape mutants were observed upon serial passaging. Our study provides the first proof-of-concept, demonstrating that the CRISPR/Cas9 system can be efficiently used to block MDV replication. The presented findings lay the foundation for future research to completely protect chickens from this deadly pathogen.


Bladder surface glycosaminoglycans: an epithelial permeability barrier.
Sulfated polysaccharide's ability to modulate the movement of small molecules was examined both in vivo and in vitro. For the in vivo test, the rabbit bladder was utilized and C-14 labeled urea 45-Ca, or 3H2O was placed into the lumen of control bladders, bladders pretreated with protamine sulfate (20 mg./cc) and bladders pretreated with protamine sulfate (20 mg./cc) plus pentosanpoly-sulfate (PPS), 10 mg./cc. After 45 minutes, the controls absorbed 21% of the urea, 16% of the calcium, and 38% of the 3H2O; the protamine treated group 40% urea, 23% calcium, and 51% H2O; the PPS only group 22% urea and the protamine plus PPS group absorbed 24% of urea, 18% calcium, and 44% water. Differences between the control and protamine groups were statistically significant, p less than 0.01 for urea 45-Ca and 3H2O. The bladder mucosa contained a significantly higher concentration of urea and calcium after protamine treatment which were both reversed by PPS (p less than 0.01) while 3H2O content went down significantly (p = 0.03), reflecting a loss of the hydrophilic effect of bladder GAG. The control mucosas had 250 cpm/mg. tissue urea for Ca 64 cpm/mg. and water 262 cpm/mg., the protamine group urea 498 cpm/mg., Ca 190 cpm/mg., and H2O 139 cpm/mg.; the protamine plus PPS group urea 344 cpm/mg., Ca 129 cpm/mg., and water 168 cpm/mg. For the in vitro studies, an Ussing chamber was employed. Normal rabbit bladder membranes were placed in the chambers and the potential difference was zeroed across the membrane. There were three groups, membranes that were treated only with the irrigating solution, membranes pretreated with protamine, and membranes pretreated with protamine plus PPS. At the end of 40 minutes, there was an approximately 1.2% movement of urea across the control membrane, a 3.5% movement across the protamine treated membrane (a significant increase p less than 0.001) and a 1.1% movement across the protamine plus PPS treated membrane. It would appear that the surface polysaccharide may play an important role as a bladder permeability barrier in modulating both charged and uncharged small molecule movement in that its ability to impair such movement can be inhibited by protamine and this protamine effect can be reversed by a treatment with an exogenous sulfated polysaccharide.


In vivo transcription studies of coliphage 186.
The temporal apperance of transcripts from the 186 chromosome has been determined by pulse-labeling at different times after prophage induction and hybridization of RNA extracts to cloned restriction fragments of 186. Studies with different mutants and induction in the presence of chloramphenicol suggested a controlled pattern of transcription and led us to propose the existence of a primary control gene analogous to the lambda gene N.


Integration of polyaniline/poly(acrylic acid) films and redox enzymes on electrode supports: an in situ electrochemical/surface plasmon resonance study of the bioelectrocatalyzed oxidation of glucose or lactate in the integrated bioelectrocatalytic systems.
Electropolymerization of aniline in the presence of poly(acrylic acid) on Au electrodes yields a polyaniline/poly(acrylic acid) composite film, exhibiting reversible redox functions in aqueous solutions at pH = 7.0. In situ electrochemical-SPR measurements are used to identify the dynamics of swelling and shrinking of the polymer film upon the oxidation of the polyaniline (PAn) to its oxidized state (PAn(2+)) and the reduction of the oxidized polymer (PAn(2+)) back to its reduced state (PAn), respectively. Covalent attachment of N(6)-(2-aminoethyl)-flavin adenin dinucleotide (amino-FAD, 1) to the carboxylic groups of the composite polyaniline/poly(acrylic acid) film followed by the reconstitution of apoglucose oxidase on the functional polymer yields an electrically contacted glucose oxidase of unprecedented electrical communication efficiency with the electrode: electron-transfer turnover rate approximately 1000 s(-1) at 30 degrees C. In situ electrochemical-SPR analyses are used to characterize the bioelectrocatalytic functions of the biomaterial-polymer interface. The current responses of the bioelectrocatalytic system increase as the glucose concentrations are elevated. Similarly, the SPR spectra of the system are controlled by the concentration of glucose. The glucose concentration controls the steady-state concentration ratio of PAn/PAn(2+) in the film composition. Therefore, the SPR spectrum of the film measured upon its electrochemical oxidation is shifted from the spectrum typical for the oxidized PAn(2+) at low glucose concentration to the spectrum characteristic of the reduced PAn at high glucose concentration. Similarly, the polyaniline/poly(acrylic acid) film acts as an electrocatalyst for the oxidation of NADH. Accordingly, an integrated bioelectrocatalytic assembly was constructed on the electrode by the covalent attachment of N(6)-(2-aminoethyl)-beta-nicotinamide adenine dinucleotide (amino-NAD(+), 2) to the polymer film, and the two-dimensional cross-linking of an affinity complex formed between lactate dehydrogenase and the NAD(+)-cofactor units associated with the polymer using glutaric dialdehyde as a cross-linker. In situ electrochemical-SPR measurements are used to characterize the bioelectrocatalytic functions of the system. The amperometric responses of the system increase as the concentrations of lactate are elevated, and an electron-transfer turnover rate of 350 s(-1) between the biocatalyst and the electrode is estimated. As the PAn(2+) oxidizes the NADH units generated by the biocatalyzed oxidation of lactate, the PAn/PAn(2+) steady-state ratio in the film is controlled by the concentration of lactate. Accordingly, the SPR spectrum measured upon electrochemical oxidation of the film is similar to the spectrum of PAn(2+) at low lactate concentration, whereas the SPR spectrum resembles that of PAn at high concentrations of lactate.


Effects of Lifestyle Intervention towards Obesity and Blood Pressure among Housewives in Klang Valley: A Quasi-Experimental Study.
The obesity rate in Malaysia is the highest in Asia. Half its population is obese or overweight. The present study aims to determine the effects of lifestyle intervention on weight loss and blood pressure among Malaysian overweight and obese housewives in Klang Valley. A quasi-experimental study with 328 obese and overweight low socio- economic status housewives aged 18-59 years old who met the screening criteria participated in the study. They were recruited into an intervention group (N = 169) or control group (N = 159). The intervention group received a lifestyle intervention consisting of a diet, physical activity and self-monitoring behavior package. The control group (delayed intervention group) received a women's health seminar package. Both groups were followed up for six months. Weight, body mass index (BMI), and blood pressure were evaluated both pre- and post-intervention. A total of 124 participants from the intervention group and 93 participants from the control group completed the study. Mean weight loss was 1.13 ± 2.70 kg (P < 0.05) in the intervention group and 0.97 ± 2.60 kg (P < 0.05) in the control group. Systolic blood pressure (SBP) reductions in the intervention group were 5.84 ± 18.10 mmHg (P < 0.05). The control group showed reduction in SBP 6.04 ± 14.52 mmHg (P < 0.05). Both group had non-significant DBP reduction. Multivariate analysis via General Linear Model Repeated Measures observed no significant differences in terms of parameter changes with time in both groups for all parameters. The results indicate that the lifestyle interventions in this study resulted in modest weight loss and thus decreased BMI and blood pressure (SBP) within six months of intervention.


Efflux of dopamine from the synaptic cleft in the nucleus accumbens of the rat brain.
Synaptic release of dopamine in the nucleus accumbens of the intact rat brain elicited by a single electrical impulse applied to ascending dopaminergic fibers results in extracellular concentrations sufficient to bind the known dopamine receptors. The dopamine concentration observed after four rapid, sequential pulses is exactly four times greater and is unaffected by pharmacological antagonism of dopamine uptake and receptor sites at supramaximal concentrations. Thus, dopamine efflux from the synaptic cleft is not restricted by binding to intrasynaptic proteins on the time scale of the measurements (50-100 msec). The extracellular concentration, as a result of a single stimulus pulse, is 0.25 microM and is rapidly removed by extrasynaptic uptake. This maximal, transient concentration of dopamine is 60 times higher than steady-state concentrations reported previously using dialysis techniques, illustrating that dopamine extracellular concentrations are spatially and temporally heterogenous. In contrast to ACh transmission at the neuromuscular junction, the dopamine synapse in the telencephalon is designed for the effective efflux of dopamine from the synaptic cleft to the extrasynaptic compartment during neurotransmission.


An evaluation of the effectiveness of teaching anatomy to rheumatologists through combined musculoskeletal sonoanatomy and human cadaveric dissection.
Our aim was to evaluate the effectiveness of teaching anatomy through combined musculoskeletal sonoanatomy and human cadaveric dissection for rheumatologists practising musculoskeletal US. The principal focus was on scanning and then dissecting relevant musculoskeletal structures. Outcomes measured included confidence levels and objective knowledge. A mixed-methods approach of evaluation and descriptive statistical data analysis was performed. The change in confidence ratings by delegates after the teaching event as represented by the mean difference (s.d.) (s.e.m.) for identification of surface anatomy was 1.846 (1.281) (0.355), with Student's paired t = 5.196 and P=0.000223. The mean difference (s.d.) (s.e.m) for performing IA injections was 1.538 (1.266) (0.351), with Student's paired t = 4.382, P=0.001, and for recognizing sonoanatomical structures it was 1.769 (1.235) (0.343), with Student's paired t = 5.165 and P= 0.000235. There was a significant increase in correct identification of anatomical and sonoanatomical knowledge in the pre- and post-course assessments. Rotator cuff interval region improved from 13 to 73%, P = 0.004; knee tendons insertion sites from 47 to 93%, P = 0.016; and muscles not adjacent to joints from 27 to 93%, P = 0.002. Dissection of joints enabled a three-dimensional relational mind map of the relevant regions of the human body, producing clarity in understanding regional relational topographic anatomy and sonoanatomy. The combination of US and cadaveric dissection improved learners' satisfaction, confidence and knowledge in areas where soft tissue complaints are common, which is likely to lead to accurate early diagnosis and cost-conscious, better overall care.


Tunable synthesis of metal-graphene complex nanostructures and their catalytic ability for solvent-free cyclohexene oxidation in air.
A one-step method was developed for the controllable construction of metal-graphene core-shell structures, hollow graphene nanospheres, and a high density of metal nanoparticles supported on graphene. The metal-graphene core-shell nanostructures as nanocatalysts show excellent catalytic ability for the selective oxidation of cyclohexene.


Water loss regulation in mature Hevea brasiliensis: effects of intermittent drought in the rainy season and hydraulic regulation.
Effects of soil and atmospheric drought on whole-tree transpiration (E(T)), leaf water potential (Ψ(L)) and whole-tree hydraulic conductance (K(T)) were investigated in mature rubber trees (Hevea brasiliensis, clone RRIM 600) during the full canopy stage in the rainy season in a drought-prone area of northeast Thailand. Under well-watered soil conditions, transpiration was tightly regulated in response to high evaporative demand, i.e., above reference evapotranspiration (ET(0)) ~2.2 mm day(-1) or maximum vapor pressure deficit ~1.8 kPa. When the trees experienced intermittent soil drought E(T) decreased sharply when relative extractable water in the top soil was < 0.4. The midday leaf water potential (Ψ(md)) on sunny days did not change as a function of soil drought and remained stable at approximately - 1.95 MPa, i.e., displaying isohydric behavior. The decrease in E(T) was mainly due to the change in K(T). K(T) remained constant over a wide range of environmental conditions and decreased sharply at low soil water availability. A simple hydraulic model incorporating critical minimum water potential and the response of whole-tree hydraulic conductance to relative extractable water correctly simulated patterns of transpiration over 6 months. We conclude that an explicit and simplified framework of hydraulic limitation hypothesis was sufficient to describe water use regulation of a mature rubber tree stand in water-limited conditions. Given the complexity of constraints in the soil-plant-atmosphere pathway, our results confirm the relevance of this approach to synthesize the overall behavior of trees under drought.


A comparison of the behavioral and anatomical outcomes in sub-acute and chronic spinal cord injury models following treatment with human mesenchymal precursor cell transplantation and recombinant decorin.
This study assessed the potential of highly purified (Stro-1(+)) human mesenchymal precursor cells (hMPCs) in combination with the anti-scarring protein decorin to repair the injured spinal cord (SC). Donor hMPCs isolated from spinal cord injury (SCI) patients were transplanted into athymic rats as a suspension graft, alone or after previous treatment with, core (decorin(core)) and proteoglycan (decorin(pro)) isoforms of purified human recombinant decorin. Decorin was delivered via mini-osmotic pumps for 14 days following sub-acute (7 day) or chronic (1 month) SCI. hMPCs were delivered to the spinal cord at 3 weeks or 6 weeks after the initial injury at T9 level. Behavioral and anatomical analysis in this study showed statistically significant improvement in functional recovery, tissue sparing and cyst volume reduction following hMPC therapy. The combination of decorin infusion followed by hMPC therapy did not improve these measured outcomes over the use of cell therapy alone, in either sub-acute or chronic SCI regimes. However, decorin infusion did improve tissue sparing, reduce spinal tissue cavitation and increase transplanted cell survivability as compared to controls. Immunohistochemical analysis of spinal cord sections revealed differences in glial, neuronal and extracellular matrix molecule expression within each experimental group. hMPC transplanted spinal cords showed the increased presence of serotonergic (5-HT) and sensory (CGRP) axonal growth within and surrounding transplanted hMPCs for up to 2 months; however, no evidence of hMPC transdifferentiation into neuronal or glial phenotypes. The number of hMPCs was dramatically reduced overall, and no transplanted cells were detected at 8 weeks post-injection using lentiviral GFP labeling and human nuclear antigen antibody labeling. The presence of recombinant decorin in the cell transplantation regimes delayed in part the loss of donor cells, with small numbers remaining at 2 months after transplantation. In vitro co-culture experiments with embryonic dorsal root ganglion explants revealed the growth promoting properties of hMPCs. Decorin did not increase axonal outgrowth from that achieved by hMPCs. We provide evidence for the first time that (Stro-1(+)) hMPCs provide: i) an advantageous source of allografts for stem cell transplantation for sub-acute and chronic spinal cord therapy, and (ii) a positive host microenvironment that promotes tissue sparing/repair that subsequently improves behavioral outcomes after SCI. This was not measurably improved by recombinant decorin treatment, but does provide important information for the future development and potential use of decorin in contusive SCI therapy.


Hybridization between the African clawed frogs Xenopus laevis and Xenopus muelleri (Pipidae) increases the multiplicity of antimicrobial peptides in skin secretions of female offspring.
Peptidomic analysis was used to compare the distribution of host-defense peptides in norepinephrine-stimulated skin secretions from laboratory-generated female F1 hybrids of the common clawed frog Xenopus laevis (Daudin, 1802) and Mueller's clawed frog Xenopus muelleri (Peters, 1844) with the corresponding distribution in skin secretions from the parent species. A total of 18 peptides were identified in secretions from the hybrid frogs. Eleven peptides (magainin-1, magainin-2, CPF-1, CPF-3, CPF-4, CPF-5, CPF-6, CPF-7, XPF-1, XPF-2, and PGLa) were identified in secretions of both the hybrids and X. laevis. Four peptides (magainin-M1, XPF-M1, CPF-M1, and tigerinin-M1) were previously found in skin secretions of X. muelleri but magainin-M2 and CPF-M2 from X. muelleri were not detected. Three previously undescribed peptides (magainin-LM1, PGLa-LM1, and CPF-LM1) were purified from the secretions of the hybrid frogs that were not detected in secretions from either X. laevis or X. muelleri. Magainin-LM1 differs from magainin-2 from X. laevis by a single amino acid substitution (Gly(13)→Ala) but PGLa-LM1 and CPF-LM1 differ appreciably in structure from orthologs in the parent species. CPF-LM1 shows potent, broad-spectrum antimicrobial activity and is hemolytic. The data indicate that hybridization increases the multiplicity of skin host-defense peptides in skin secretions. As the female F1 hybrids are fertile, hybridization may represent an adaptive strategy among Xenopus species to increase protection against pathogenic microorganisms in the environment.


Lachrymal concentration of norfloxacin after a single ocular instillation in humans.
Lachrymal concentrations of the fluoroquinolone norfloxacin were studied in 30 subjects (aged 24-35 years) after a single ocular instillation (0.3% solution, 50 ul) in comparison with ofloxacin instilled at the same dose. Lachrymal levels were measured by high performance liquid chromatography (HPLC). Lachrymal peak levels were comparable 5 min after instillation but norfloxacin appeared to have a longer elimination half-life than ofloxacin. The AUC (area under the curve) for norfloxacin was therefore higher than for ofloxacin (35.9 and 10.7 mg* min/ml, respectively). These data suggest that norfloxacin may have higher corneal retention than ofloxacin, so this antibiotic may be indicated in superficial infections dependent on fluoroquinolone-sensitive bacteria.


PIMEX--an instructive way to study vibration exposure and work posture.
The PIMEX-method (PIcture-Mix-EXposure) involves measurement of exposure with a direct-reading instrument. The signal from the instrument is superimposed to the recording from a video camera to produce a video film which continuously shows the subject at work and how exposure varies. Application can be a physical factor such as vibration. We used this new method to study vibrations from hand held grinders. Using Brüel and Kjaer miniaturized accelerometer 4374 and vibration meter 2513 we measured vibrations at the grinders main and support handles. We studied different grinders at work on the metal surface as well as at idle speed. Workers posture such as arms in under-up position was evaluated. The PIMEX-method has been encouraging to show correlation between vibration exposure, work technique and different grinders.


Immunological and chemical studies of the streptococcal group O protein type antigens.
Two antigens present in group O streptococci have been described and are designated as type antigens. No other antigens are present in extracts of group O cells in significant quantity. Antigen I is a cell wall protein, and II is a nucleoprotein of undetermined cell location. The immunological specificity of the latter resides only in the protein. Both antigens have been extracted from the cell with water or 0.2 n HCl (pH 2) at 100 C, and antigen I can be removed by trypsin. After purification, pH 2 was found to destroy the serological activity of each antigen. Antgens I and II and also the O polysaccharide antigen can be distinguished on a single agar-gel diffusion plate with group O whole-cell antiserum. N- and C-terminal amino acid analyses and total amino acid composition prove that I and II are separate molecular species. Each possesses properties which separate them from the streptococcal M, R, and T proteins. Antigen I separated into three fractions, each possessing serological specificity, on a diethylaminoethyl-Sephadex column at pH 8.6 during an increase in buffer molarity. The protein is considered to possess a multiple unit structure. The results indicate that the O streptococci are limited to two serological types. Group O whole-cell antisera, after adsorption with a strain not containing the O polysaccharide, can now be prepared for the positive identification of the types of group O streptococci.


Structure and conformation of the branch core triribonucleotide containing 2'-5' and 3'-5' phosphodiester linkages (A 2'p5'G 3'p5'C) i solution, essential for yeast mRNA splicing, deduced from 1H-NMR.
The non-exchangeable 1H-NMR signals of the branch core trinucleotide of the lariat branch site (A2'p5'G3'p5'C, 1) and its derivatives 2 and 3 are completely assigned using one- and two-dimensional NMR techniques including NOE, COSY, NOESY, 1H-1H INADEQUATE and 2D-J-resolved spectroscopy. From the vicinal coupling constants in the individual ribose rings, NOE data and T1 measurements, the following properties of the trimers are deduced. (i) The unique stacking behavior of the trimers is S2'N3'N, and the sugar rings exist predominantly in the N-conformation (3'-endo-2'-exo). (ii) The sugar-base orientations appear to be anti. (iii) The branched trimers exist in solution as single-stranded right-handed conformations resembling A-RNA with stacking between the adenine and guanine residues in aqueous solution at 21 degrees C and pH 7.2. (iv) The calculated values for the torsion angles epsilon t and gamma+ for the trimers are 201-203 degrees and 71-86%, respectively, while the percent beta t values are higher for the guanine (87-92%) than the cytosine residues (73-77%). The computer generated depiction of the triribonucleotide 1 is also shown. These subtle structural features may act as recognition signals for this critical lariat branch site which is essential for the second step in yeast mRNA splicing.


Correlation Between TCF7+ T Cells and Prognosis of Patients With Oral Squamous Cell Carcinoma.
To investigate whether TCF7+ T cells constitute an important factor to improve the existing postoperative prediction model for patients with oral squamous cell carcinoma. TCF7+ T cells were detected in the tissues of 167 OSCC patients by multiplex immunofluorescence. The percentage of TCF7+ T cells was transformed into a dichotomous variable, combined with the clinicopathological data for the OSCC patients, and then subjected to univariate and multivariate analyses. The derived independent predictors were then incorporated into risk models to analyze their relationship with the prognosis of patients. The high TCF7+ group had a better prognosis than the low TCF7+ group (OS: p<0.001; RFS: p<0.001). Univariate and multivariate analyses showed that TCF7+ T cells serve as an independent predictor of OSCC (univariate/multivariate analysis: p<0.001). In Cox risk progression models, inclusion of the TCF7+ T cell percentage improved the predictive accuracy of Grade and TNM stage (Grade-OS/RFS: p<0.001; TNM-OS/RFS: p<0.001; TNM+Grade-OS: p<0.001, TNM+Grade-RFS: p=0.004). Inclusion of the TCF7+ T cell percentage improved the clinical utility. TCF7+ T cells can act as an independent predictor for postoperative OSCC patients. The inclusion of TCF7+ T cells improved the predictive accuracy and clinical utility of the nomograms to different degrees.


Benign familial nocturnal alternating hemiplegia of childhood.
In infancy, two brothers developed recurrent attacks of alternating or bilateral hemiplegia arising exclusively out of sleep. The episodes were terminated by even brief sleep. Neither child had hypotonia, dystonic attacks, paroxysmal eye movement abnormalities, or other features characteristic of the now-classic form of alternating hemiplegia of childhood (AHC). The development of the brothers has so far remained normal. Both parents have a history of migraine. In the older boy, magnetic resonance spectroscopy (MRS) of muscle showed increased inorganic phosphate similar to what is found in children with AHC. In the younger brother and parents, MRS of muscle was normal. Other investigations were unrevealing. Flunarizine greatly reduced the duration of attacks. This genetically determined disorder represents a specific entity that is probably migraine-related and is easily misdiagnosed as AHC. Because of its benign course, particularly as far as mental development is concerned, it must be distinguished from classic AHC, which has a terrible prognosis.


Evaluation of hepatitis C virus serotypes and IgM anti-HCV antibodies in two groups of anti-HCV positive subjects in south-east Italy.
We investigated the relation between the presence of IgM antibodies to hepatitis C virus (HCV) and serotypes of HCV, in particular when a high level of IgM antibodies are present in patients infected with HCV serotype I and may be associated with the source, duration and evolution of infection. The study involved two anti-HCV positive groups with chronic liver disease from the same area, one of which was a group of haemodialysis patients attending the same haemodialysis centre.


DEGRO practical guideline for partial-breast irradiation.
This consensus statement from the Breast Cancer Working Group of the German Society for Radiation Oncology (DEGRO) aims to define practical guidelines for accelerated partial-breast irradiation (APBI). Recent recommendations for relevant aspects of APBI were summarized and a panel of experts reviewed all the relevant literature. Panel members of the DEGRO experts participated in a series of conferences, supplemented their clinical experience, performed a literature review, and formulated recommendations for implementing APBI in clinical routine, focusing on patient selection, target definition, and treatment technique. Appropriate patient selection, target definition for different APBI techniques, and basic rules for appropriate APBI techniques for clinical routine outside of clinical trials are described. Detailed recommendations for APBI in daily practice, including dose constraints, are given. Guidelines are mandatory to assure optimal results of APBI using different techniques.


Characterizing the molecular mechanisms of acquired temozolomide resistance in the U251 glioblastoma cell line by protein microarray.
Acquired chemoresistance refers to tumor cells gradually losing their sensitivity to anticancer drugs during the course of treatment, resulting in tumor progression or recurrence. This phenomenon, which has deleterious outcomes for the patient, has long been observed in patients with glioblastoma receiving temozolomide (TMZ)-based radiochemotherapy. Currently, the mechanisms for acquired TMZ chemoresistance are not fully understood. In the present study, a TMZ-resistant cell line U251R with a 4-fold 50% inhibition concentration compared with its TMZ-sensitive parent cell line was isolated by incremental long-time TMZ treatment in the human glioblastoma cell line U251. Fluorescence-activated cell sorting analysis indicated G2/M arrest and a lower proportion of cells in the S phase, accompanied by a decreased apoptosis rate in the U251R cell line compared with the parental U251 cell line. In addition, a sphere-formation assay indicated an increased self-renewal capacity in U251R cells. Furthermore, a high-throughput protein microarray unveiled more than 200 differentially expressed proteins as potential molecular targets accounting for acquired TMZ resistance. Subsequent bioinformatics analysis illustrated the molecular and signaling networks and revealed the central role of SRC. Immunoblotting and reverse-transcription quantitative polymerase chain reaction analysis further confirmed the expressional upregulation of SRC family kinases. Moreover, SRC knockdown led to partial reversal of TMZ resistance in the U251R cell line and sensitization in the U373 cell line. These data helped to develop a comprehensive understanding of survival strategies, particularly with respect to pro-stemness regulation, which could be potential targets for overcoming TMZ resistance.


Mid-upper arm circumference in hospitalized pediatric patients with cancer: Is there an association with malnutrition?
Malnutrition is common in children/adolescents with cancer. Mid-upper arm circumference (MUAC) is recommended by the International Society of Pediatric Oncology for measuring nutrition status. The aim was to evaluate the nutrition status of pediatric patients with cancer and verify whether MUAC is associated with malnutrition and length of stay (LOS). This was an observational study of patients aged 0-19 years with solid and hematological neoplasms. Data included anthropometric measurements of nutrition status by body mass index/age (BMI/A), weight/age, weight/height, height/age, MUAC, mid-arm muscle circumference, triceps skinfold thickness (TSF), and biochemical measurements of characterizing inflammation (serum prealbumin level, serum albumin level, and C-reactive protein [CRP]). Statistical tests such as the Student t test, Mann-Whitney test, and Spearman correlation were employed to analyze the data. Logistic regression was used for multivariate analysis. Of 111 patients, 51.7% (n = 57) were male, and solid tumors were prevalent (76.6%, n = 85). The median age was 8.5 years. The anthropometric measurements of the arm revealed a higher prevalence of malnutrition when compared with anthropometric indices. According to the Spearman correlation, a high and positive correlation was observed between MUAC and BMI/A (ρ = 0.800 and P < 0.001). There was an association between MUAC, TSF, and high CRP with LOS (P = 0.025, P = 0.003, and P < 0.001, respectively). Patients with a MUAC (less than the fifth percentile) had almost 2.73 times the chance of having a prolonged LOS. MUAC was a good parameter for the classification of nutrition status and the assessment of LOS in pediatric patients with cancer.


Polyamine deprivation stimulates natural killer cell activity in cancerous mice.
It has recently been established that the total blockade of all endogenous and exogenous sources of polyamines by a drug containing polyamine deficient chow (DC-PDC+) could inhibit tumor growth in vivo and increase the antitumoral efficacy of chemotherapy drugs. We show here that polyamine deprivation obtained with DC-PDC+ not only influences tumor development via reduction of polyamine concentrations in the tumor itself but, in addition, stimulates cells of the non-specific immune system specialized in tumor cell killing. We report that mice grafted with the 3LL carcinoma present a dramatic decrease in the cytotoxic activity of their natural killer (NK) cells. When these animals are treated with DC-PDC+, their NK cell activity is completely restored to normal values. Normalization of leucocyte number and differential count was observed as well. With respect to the different components of the DC-PDC+, it was observed that the endogenous and exogenous sources of polyamines have a different degree of impact on tumor development. For example, when only polyamines from digestive sources are deprived, a weak but significant improvement in NK activity and antitumoral effects were observed, without affecting intra-tumoral polyamine concentrations. We conclude that polyamines, secreted by the tumor itself as well as absorbed through the gastrointestinal tract, could now be considered not only as autocrine growth factors but also as natural immunosuppressive factors.


Beneficial antipsychotic effects of celecoxib add-on therapy compared to risperidone alone in schizophrenia.
Abnormalities in the immune system in schizophrenia have been described. However, important findings such as high levels of activating cytokines in the CSF and signs of CNS inflammation have been controversial. The authors conducted a trial of the new selective cyclooxygenase-2 inhibitor celecoxib, an immunomodulatory drug, in schizophrenic patients to evaluate its therapeutic effects. In a prospective, double-blind evaluation, 50 patients with an acute exacerbation of schizophrenia were randomly assigned to either risperidone plus celecoxib or risperidone plus placebo. After a washout period, 25 patients received 2-6 mg/day of risperidone plus placebo and 25 received risperidone plus 400 mg/day of celecoxib for 5 weeks. The treatment effect was calculated by analysis of covariance. There were no significant differences between groups in age, sex, duration or severity of disease or psychopathology, or risperidone dose or plasma level. Over 5 weeks, both groups of patients showed significant improvement in scores on the Positive and Negative Syndrome Scale and on all subscales. However, the celecoxib group showed significantly greater improvement in the total score. Additional treatment with celecoxib has significant positive effects on the therapeutic action of risperidone with regard to total schizophrenia psychopathology. Moreover, the fact that treatment with an immunomodulatory drug showed beneficial effects on schizophrenia symptoms indicates that immune dysfunction in schizophrenia is not just an epiphenomenon but is related to the pathomechanism of the disorder. However, a nonimmunological therapeutic effect of celecoxib mediated by the N-methyl-D-aspartic acid receptor has to be taken into account.


Linking Retinal Microvasculature Features With Severity of Diabetic Retinopathy Using Optical Coherence Tomography Angiography.
To correlate retinal vascular features with severity and systemic indicators of diabetic retinopathy (DR) using optical coherence tomography angiography (OCTA). A total of 209 eyes of 122 type 2 diabetes mellitus patients with DR and 60 eyes of 31 normal Indian subjects underwent OCTA imaging. The diabetic retinopathy patients were graded as having either nonproliferative diabetic retinopathy (NPDR: mild, moderate, and severe NPDR using Early Treatment Diabetic Retinopathy Study classification) or proliferative diabetic retinopathy (PDR). Local fractal analysis was applied to the superficial and deep retinal OCTA images. Foveal avascular zone area (FAZ in mm2); vessel density (%); spacing between large vessels (%); and spacing between small vessels (%) were analyzed. Sensitivity and specificity of vascular parameters were assessed with receiver operating characteristics (ROC) curve. Normal eyes had a significantly lower FAZ area, higher vessel density, and lower spacing between large and small vessels compared with DR grades (P < 0.001). In the superficial layer, PDR and severe NPDR had higher spacing between large vessels than mild and moderate NPDR (P = 0.04). However, mild NPDR had higher spacing between the small vessels (P < 0.001). Spacing between the large vessels in the superficial retinal layer correlated positively with HbA1c (r = 0.25, P = 0.03); fasting (r = 0.23, P = 0.02); and postprandial (r = 0.26, P = 0.03) blood sugar. The same spacing in the deep retinal vascular plexus had the highest area under the ROC curve (0.99 ± 0.01) and was uniformly elevated in all diabetic eyes (P > 0.05). Spacing between the large vessels in the superficial and deep retinal layers had superior diagnostic performance than overall vessel density.


Androgen deprivation therapy: a cornerstone in the treatment of advanced prostate cancer.
Decreased incidence of prostate cancer, improved therapies, and early detection have all have played some role in the decrease in prostate cancer mortality. The author discusses the development of improved methods of androgen deprivation therapy and demonstrates its significance in improved management of prostatic carcinoma.


[Significance of miR-155, miR-34a and miR-30a expression in diffuse large B-cell lymphoma].
To determine the expressions of miR-155, miR-34a and miR-30a in diffuse large B-cell lymphoma (DLBCL) and to explore their potential correlation with clinicopathological characteristics. The expression level of miR-155, miR-34a and miR-30a in 46 DLBCL samples were determined with TaqMan real-time polymerase chain reaction. Interphase fluorescence in situ hybridization (I-FISH) was performed to detect MYC and p53 genes' status, and immunohistochemistry (Envision method) was used to evaluate the expression of CD3, CD10, CD20, BCL-6 and MUM-1 in DLBCL. The DLBCLs were classified into germinal center B cell-like (GCB) and non germinal center B cell-like (non-GCB) subtypes according to Hans' criteria. Compared with normal controls, miR-155 expression level was significantly higher in DLBCL. The expression level of miR-155 in non-GCB type was higher than that in GCB type. It was shown that the patients with MYC rearrangement had lower expression level of miR-155 than the negative controls. Compared with p53 normal group, the expression level of miR-34a was significantly lower in p53 deletion group. It was also shown that the patients with BCL-6 protein expression had lower expression of miR-30a compared with the negative group. miR-155 expression level is different in normal controls, DLBCL and patients with subtype DLBCL. It therefore has a diagnosis value for DLBCL. miR-34a is of great prognostic significance. miR-155, miR-34a and miR-30a may be potential therapy targets for DLBCL.


Riboflavin depletion of intestinal cells in vitro leads to impaired energy generation and enhanced oxidative stress.
Riboflavin is an essential component of the human diet, with an established role for its derivative cofactors in oxidative metabolism. Our previous in vivo data suggest that riboflavin may act as a signalling molecule in the intestinal lumen, regulating crypt development and cell turnover. Our in vitro studies in riboflavin-depleted intestinal cells in culture indicate that riboflavin depletion impairs normal mitosis. The aim of the study was to establish an improved intestinal cell model of riboflavin depletion using the structural analogue of riboflavin, lumiflavin (7,8,10-trimethyl-isoalloxazine) and to determine effects on cell function. The study was conducted using three intestinal cell lines, Caco-2, HCT116 and HT29 cells. Cell growth was inhibited in all three cell lines, in a lumiflavin concentration-dependent manner. Riboflavin depletion was confirmed through a significant decrease in intracellular riboflavin concentrations in Caco-2 and HT29 cell lines and a significant increase in the activation coefficient for the flavin adenine dinucleotide-dependent enzyme glutathione reductase. Riboflavin depletion led to a significant reduction in intracellular ATP concentration, and an enhanced generation of reactive oxygen species was also observed in response to riboflavin depletion, in all cell lines; effects were at least fivefold greater in Caco-2 cells than other cells. Riboflavin-depleted Caco-2 and HCT116 cells also showed an irreversible loss of proliferative potential. A model system of intracellular riboflavin depletion in intestinal epithelial cells has been developed. Riboflavin depletion induced by lumiflavin results in oxidative stress and a disruption of energy generation, which may contribute to observed effects on cell proliferation.


Formation of a Reactive, Alkyl Thiolate-Ligated FeIII-Superoxo Intermediate Derived from Dioxygen.
Herein, we describe an alkyl thiolate-ligated iron complex that reacts with dioxygen to form an unprecedented example of an iron superoxo (O2•-) intermediate, [FeIII(S2Me2N3(Pr,Pr))(O2)] (4), which is capable of cleaving strong C-H bonds. A cysteinate-ligated iron superoxo intermediate is proposed to play a key role in the biosynthesis of β-lactam antibiotics by isopenicillin N-synthase (IPNS). Superoxo 4 converts to a metastable putative Fe(III)-OOH intermediate, at rates that are dependent on the C-H bond strength of the H atom donor, with a kinetic isotope effect ( kH/ kD = 4.8) comparable to that of IPNS ( kH/ kD = 5.6). The bond dissociation energy of the C-H bonds cleaved by 4 (92 kcal/mol) is comparable to C-H bonds cleaved by IPNS (93 kcal/mol). Both the calculated and experimental electronic absorption spectra of 4 are comparable to those of the putative IPNS superoxo intermediate, and are shown to involve RS- → Fe-O2•- and O2•- → Fe charge transfer transitions. The π-back-donation by the electron-rich alkyl thiolate presumably facilitates this reactivity by increasing the basicity of the distal oxygen. The frontier orbitals of 4 are shown to consist of two strongly coupled unpaired electrons of opposite spin, one in a superoxo π*(O-O) orbital, and the other in an Fe(d xy) orbital.


Application of Carbon Nanotubes in Breast Cancer Therapy.
Conjugated single-walled carbon nanotubes (SWNT) have been shown to be promising in cancer-targeted accumulation and is biocompatible, easily excreted, and possesses little toxicity. The present study aims at reviewing the recent advancements in carbon nanotubes especially SWNT for improving the treatment of breast cancer. Nanotube drug delivery system is a potential high efficacy therapy with minimum side effects for future tumor therapy with low doses of drug.


Investigation of the effects of experimental cryptorchidism formed in prepubertal rats during the postpubertal and adult periods.
Investigation of the effects of experimental cryptorchidism formed in prepubertal rats during the postpubertal and adult periods. The abdominal cavities of 20 rats in the control group were just opened and closed without any further surgical procedure. The testicles of 20 rats in the experimental group were fixed to the anterolateral wall of the abdominal cavity under general anesthesia. At the 6th month, blood and testis tissue samples of rats were obtained and the rats were sacrificed and the experiment was terminated. Statistical analysis was done by SPSS 15.0 package software. The histopathological evaluation of testicular tissues of rats in the control group was in the normal limits. Of the experimental group, the testicular tissues had injury in nearly half of the rats, in which the testicles were fixed in the abdomen for three months that is the 1st subgroup. FSH levels were statistically significantly elevated in both experimental groups of rats (p<0.01). Cryptorchidism formed experimentally causes a partial damage in the testicular tissue of pubertal rats, however the damage increases gradually in the testicles of rats, in which the testicles were descended and left in the scrotum for another three months. The statistically elevated levels of FSH in both experimental groups of rats can be accounted for severe testicular damage. The timing in descending the undescended testicles to the scrotum is essential and should be performed before the puberty and as early as possible in avoiding the risk of infertility (Tab. 3, Fig. 5, Ref. 44).


Ketoprofen vs etofenamate in a controlled double-blind study: evidence of topical effectiveness in soft tissue rheumatic pain.
The efficacy and safety of topical application of ketoprofen gel and etofenamate gel were studied in a controlled double-blind clinical trial. Thirty-six patients affected by inflammation of tendons, sheaths and bursae entered the study, and were treated for seven days. The parameters studied were: pain scale, Ritchie index, stiffness, pain on active and passive movement and functional capacity. The results showed that ketoprofen gel and etofenamate gel were able to induce remission of the inflammatory symptoms in soft tissue rheumatic pain. No side-effects were detected in either group.


Induction of labor--pharmacology methods.
Pharmacologic methods for induction of labor have been used for many decades. Pharmacologic agents have an advantage over mechanical methods in that they can be used during both the initial cervical ripening stage of induction and throughout the second stage of labor. Pharmacologic induction agents such as prostaglandins and oxytocin are commonly used for labor and delivery floors and are well established for use in cervical ripening. Nitric oxide donors and mifepristone are known agents in medicine but are new and actively studied in the area of cervical ripening. These agents are introduced and analyzed in this review.


Autonomous underwater adhesion driven by water-induced interfacial rearrangement.
Underwater adhesives receive extensive attention due to their wide applications in marine explorations and various related industries. However, current adhesives still suffer from excessive water absorption and lack of spontaneity. Herein, we report an autonomous underwater adhesive based on poly(2-hydroxyethyl methacrylate-co-benzyl methacrylate) amphiphilic polymeric matrix swollen by hydrophobic imidazolium ionic liquid. The as-prepared adhesive is tough and flexible, showing little to none instantaneous underwater adhesion onto the PET substrate, whereas its adhesion energy on the substrate can grow more than 5 times to 458 J·m-2 after 24 hours. More importantly, this process is entirely spontaneous, without any external pressing force. Our comprehensive studies based on experimental characterizations and molecular dynamic simulations confirm that such autonomous adhesion process is driven by water-induced rearrangement of the functional groups. It is believed that such material can provide insights into the development of next-generation smart adhesives.


Normal stresses at the gelation transition.
A simple Rouse-type model, generalized to incorporate the effects of chemical cross-links, is used to obtain a theoretical prediction for the critical behavior of the normal-stress coefficients Psi(1) and Psi(2) in polymeric liquids when approaching the gelation transition from the sol side. While the exact calculation shows Psi(2) identical with 0, a typical result for these types of models, an additional scaling ansatz is used to demonstrate that Psi(1) diverges with a critical exponent l=k+z. Here, k denotes the critical exponent of the shear viscosity and z the exponent governing the divergence of the time scale in the Kohlrausch decay of the shear-stress relaxation function. For cross-links distributed according to mean-field percolation, this scaling relation yields l=3, in accordance with an exact expression for the first normal-stress coefficient based on a replica calculation. Alternatively, using three-dimensional percolation for the cross-link ensemble we find the value l approximately 4.9. Results on time-dependent normal-stress response are also presented.


A prospective randomized trial examining the use of a closed suction drain shows no influence on strength or function in primary total knee arthroplasty.
The aim of this study was to determine whether closed suction drain (CSD) use influences recovery of quadriceps strength and to examine the effects of drain use on secondary outcomes: quadriceps activation, intra-articular effusion, bioelectrical measure of swelling, range of movement (ROM), pain, and wound healing complications. A total of 29 patients undergoing simultaneous bilateral total knee arthroplasty (TKA) were enrolled in a prospective, randomized blinded study. Patients were randomized to receive a CSD in one limb while the contralateral limb had the use of a subcutaneous drain (SCDRN) without the use of suction ('sham drain'). Isometric quadriceps strength was collected as the primary outcome. Secondary outcomes consisted of quadriceps activation, intra-articular effusion measured via ultrasound, lower limb swelling measured with bioelectrical impendence and limb girth, knee ROM, and pain. Outcomes were assessed preoperatively and postoperatively at day two, two and six weeks, and three months. Differences between limbs were determined using paired Student's t-tests or Wilcoxon's signed-rank tests. No significant differences were identified between limbs prior to surgery for the primary or secondary outcomes. No significant differences in quadriceps strength were seen between CSD and SCDRN limbs at postoperative day two (p = 0.09), two weeks (primary endpoint) (p = 0.7), six weeks (p = 0.3), or three months (p = 0.5). The secondary outcome of knee extension ROM was significantly greater in the CSD limb compared with the SCDRN (p = 0.01) at two weeks following surgery, but this difference was absent at all other intervals. Secondary outcomes of quadriceps activation, intra-articular effusion, lower limb swelling, and pain were not found to differ significantly at any timepoint following surgery. The use of CSD during TKA did not influence quadriceps strength, quadriceps activation, intra-articular effusion, lower limb swelling, ROM, or pain. These results have limited drain use by the authors in primary uncomplicated TKA. Cite this article: Bone Joint J 2019;101-B (7 Supple C):84-90.


Update on the TURBO BOOSTER spectranetics laser for lower extremity occlusive disease.
In the last two decades the endovascular treatment of peripheral arterial occlusive disease (PAOD) has gained a widespread and predominant role. New technologies have developed in the last years as atherectomy devices, self expandible nitinol stents, drug eluting devices (stent and balloons), absorbable stents. In recent years, growing interest has been dedicated to laser technology due to device improvements and literature data reporting safety and efficacy of excimer laser. The role of this new endovascular technique for the treatment of atherosclerotic arterial diseases should be considered with regard to two fields of interest: the claudicatio intermittens (CI) and the critical limb ischemia (CLI). A 20-year history with medical lasers has proven that not all lasers are equal. Lasers used and studied in the late 1980s and 1990s had poor outcomes due to inappropriate laser selection and undefined laser techniques. Over the last 10 years, multicenter studies with the excimer laser confirm that case selection, appropriate utilization of equipment, application of safe lasing techniques, and knowledge of indications and contraindications, all contribute to the successful application of laser-assisted angioplasty in complex coronary and peripheral artery disease. If applied properly, the Excimer Laser is a useful technique to transform complex obstructive arterial disease into more treatable lesions, improving the results of endovascular treatment and lowering the threshold of intervention for ''untreatable'' patients. New larger studies are requested to assess the definitive role of this technique in PAD treatment and limb salvage. This review will discuss the Laser Phisics and application in PAD along with the clinical data available to support the Excimer Laser as a reliable technology for endovascular intervention.


Evaporation of water droplets on "lock-and-key" structures with nanoscale features.
Highly ordered poly(dimethylsiloxane) microbowl arrays (MBAs) and microcap arrays (MCAs) with "lock-and-key" properties are successfully fabricated by self-assembly and electrochemical deposition. The wetting properties and evaporation dynamics of water droplets for both cases have been investigated. For the MBAs case, the wetting radius of the droplets remains unchanged until the portion of the droplet completely dries out at the end of the evaporation process. The pinning state extends for more than 99.5% of the total evaporation time, and the pinning-shrinking transition is essentially prevented whereas in the case of the MCAs the contact radius exhibits distinct stages during evaporation and the contact line retreats significantly in the middle of the evaporation process. We explain the phenomenon by a qualitative energy balance argument based on the different shrinkage types of the nanoscale-folded contact line.


Environmental and ecological factors influencing the spillover of the non-native kelp, Undaria pinnatifida, from marinas into natural rocky reef communities.
The non-native kelp, Undaria pinnatifida, is considered one of the world's worst invasive species. The northeast Atlantic is a hotspot of Undaria invasion, yet there is limited knowledge on its invasion dynamics. In the UK its distribution is strongly associated with artificial structures, primarily marina and harbour pontoons, with relatively few records of Undaria on natural substrates. Here, the southwest UK is used as a case region, to explicitly link Undaria distribution-abundance patterns in artificial marina habitats with those in natural rocky reef habitats. Using a mixture of in situ recording and video survey techniques, Undaria was found at all thirteen marina sites surveyed; but in only 17 of 35 rocky reef sites, all of which were in 2 of the 5 larger systems surveyed (Plymouth Sound and Torbay). The distribution-abundance patterns of Undaria at reef sites were analysed using zero-inflated models. The probability of finding Undaria on rocky reef increased with increasing proximity to marinas with high abundances of Undaria. Total propagule pressure from marinas also increased the probability of occurrence, and was positively related to Undaria abundance and cover at reef sites. Increases in the cover of native kelps, Laminaria spp., and wave exposure at reef sites were linked to a reduced probability of Undaria occurrence, and lower abundance and cover. Identifying high risk areas, natural boundaries and factors affecting the spread and abundance of non-native species in natural habitats is key to future management prioritisation. Where Undaria is confined to artificial substrates management may be deemed a low priority. However, the results of this study suggest that controlling the abundance and propagule pressure in artificial habitats may limit, to some extent, the spillover of Undaria into natural rocky reef habitats, where it has the potential to interact with and influence native communities.


Isolation of follicular dendritic cells from human tonsils and adenoids. II. Immunocytochemical characterization.
Follicular dendritic cells (FDC) are specialized cells found only within lymphoid follicles. They bind immune complexes and play a role in the presentation of antigen to follicular B cells and in the generation of B cell memory. In the present report the isolation of FDC from human tonsils and adenoids is described. These isolated cells have an unusual spherical arrangement and enclose lymphocytes within extensions of their membranes. Their ultrastructural features are similar to those observed in situ. The reactivity of isolated FDC with a number of monoclonal antibodies was analyzed by immunofluorescence and by immunostaining (at the electron microscopic level) with colloidal gold. In keeping with the results of previous investigations on tissue sections IgM, IgG and IgA (but not IgD) can be detected on the surface of isolated FDC, as can C3b receptors and the FDC-associated antigen detected by monoclonal antibody R4/23. The immunoglobulins associated with FDC are mostly embedded in an electron-dense material. The majority of the lymphoid cells enclosed within the membrane extensions of FDC are of B cell type. These results suggest that isolated FDC may be suitable for further in vitro investigation of their role in the humoral immune response.


Concomitant Use of Indocyanine Green Fluorescence Imaging and Interventional Radiology for Detection of Liver Segments During Laparoscopic Anatomical Liver Resection: Pilot Feasibility Study.
We investigate the feasibility of using indocyanine green (ICG) fluorescence imaging with interventional radiology (IVR) techniques for indication of liver segments during laparoscopic anatomical liver resection (LapAR). Ten patients with planned LapAR (segmentectomy V, VI, VII, or VIII) were enrolled. Both IVR and LapAR were performed in a hybrid operating room. Two dyes (indigo carmine and ICG) and embolic solutions were injected into the target liver segment. We compared imaging results during LapAR. ICG fluorescence imaging (ICGFI) provided clear visual contrast. Median contrast index value of ICGFI was 1.12, superior to that of indigo carmine (0.21; P=0.005). ICGFI was still visible during liver resection. Estimated liver resection volume and actual resected liver weight correlated significantly (R=0.906; P<0.01). ICGFI delivered by IVR provided clearer visual information than conventional indigo carmine dye, enabling precise performance of LapAR as planned (UMIN-CTR No. 000023952).


Nitric oxide inactivates the retinoblastoma pathway in chronic inflammation.
Patients with chronic inflammatory bowel disease have a high risk of colon cancer. The molecules that initiate and promote colon cancer and the cancer pathways altered remain undefined. Here, using in vitro models and a mouse model of colitis, we show that nitric oxide (NO) species induce retinoblastoma protein (pRb) hyperphosphorylation and inactivation, resulting in increased proliferation through the pRb-E2F1 pathway. NO-driven pRb hyperphosphorylation occurs through soluble guanylyl cyclase/guanosine 3',5'-cyclic monophosphate signaling and is dependent on the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase MEK/ERK and phosphatidylinositol 3-kinase/AKT pathways. Our results reveal a link between NO and pRb inactivation and provide insight into molecules that can be targeted in the prevention of the inflammation-to-cancer sequence.


Influence of body mass and height on the energy cost of running in highly trained middle- and long-distance runners.
Previous studies about the influence of body dimensions on running economy have not compared athletes specialized in different competition events. Therefore, the purpose of the present study was to assess the influence of body mass (m(b)) and height (h) on the energy cost of running (Cr) in 38 highly trained male runners, specialized in either marathon (M, n = 12), long middle-distance (5000 - 10000 m, LMD, n = 14) or short middle-distance (800 - 1500 m, SMD, n = 12), and to assess possible differences in body dimensions for each event. Subjects performed a progressive maximal exercise on the treadmill to determine oxygen uptake VO(2)) at different submaximal velocities and maximal oxygen uptake VO(2)max). Cr was calculated from VO(2) measurements. LMD runners had significantly higher mean Cr (0.192 +/- 0.007, 0.182 +/- 0.009, and 0.180 +/- 0.009 ml O(2) x kg(-1) x m(-1) for LMD, M and SMD, respectively) and VO(2)max (74.1 +/- 3.7, 68.5 +/- 2.9 and 69.7 +/- 3.4 ml x kg (-1) x min (-1)). Cr correlated with h (r = -0.86, p < 0.001) and m(b) (r = -0.77, p < 0.01) only in the SMD group. In conclusion, these data suggest that highly trained distance runners tend to show counterbalancing profiles of running economy and VO(2)max (the higher Cr, the higher VO(2) max and vice versa), and that anthropometric characteristics related with good performance are different in long-distance and middle-distance events.