Crosstalk Between Prostate Cancer Cells and Tumor-Associated Fibroblasts Enhances the Malignancy by Inhibiting the Tumor Suppressor PLZF.
Although prostate cancer is clinically manageable during the early stages of progression, metastatic progression severely compromises the prognosis and leads to mortality. Constitutive activation of STAT3 has been connected to prostate cancer malignancy, and abolishing the STAT3 activity may diminish tumor growth and metastasis. However, its suppressor genes and pathways have not been well established. In this study, we show that promyelocytic leukemia zinc finger (PLZF) has a putative tumor-suppressor function in prostate cancer by inhibiting phosphorylation of STAT3. Compared with a benign prostate, high-grade prostate cancer patient tissue was negatively correlated with PLZF expression. PLZF depletion accelerated proliferation and survival, migration, and invasion in human prostate cancer cells. Mechanistically, we demonstrated a novel role of PLZF as the transcriptional regulator of the tyrosine phosphatase SHP-1 that inhibits the oncogenic JAKs-STAT3 pathway. These results suggest that the collapse of PLZF expression by the CCL3 derived from fibroblasts accelerates the cell migration and invasion properties of prostate cancer cells. Our results suggest that increasing PLZF could be an attractive strategy for suppressing prostate cancer metastasis as well as for tumor growth.


Knockdown of JARID2 inhibits the viability and migration of placenta trophoblast cells in preeclampsia.
Protein Jumonji (JARID2) is a member of the Jumonji family of proteins and has been demonstrated to regulate cell proliferation and invasion. However, little is known about the role of JARID2 in the metastasis of placenta trophoblast cells. In the present study, the effect and the underlying molecular mechanism of JARID2 on trophoblast cell viability and invasion was investigated. The expression of JARID2 in placental tissues was analyzed by reverse transcription‑quantitative polymerase chain reaction and western blotting. HTR8/SVneo cells were transfected with si‑JARID2 or scramble for 24 h. Cell viability, migration and invasion in HTR8/SVneo cells were then evaluated. The expression levels of matrix metallopeptidase 2 (MMP2), MMP9, phosphorylated phosphatidylinositol 3‑kinase (p‑PI3K), PI3K, phosphorylated AKT serine/threonine kinase 1 (p‑Akt) and Akt in HTR8/SVneo cells were also detected using western blotting. The results of the present study demonstrated that JARID2 is underexpressed in human preeclamptic placentas. The knockdown of JARID2 significantly inhibited the viability, migration and invasion of HTR8/SVneo cells. In addition, the knockdown of JARID2 significantly decreased the levels of phosphorylated PI3K and Akt in HTR8/SVneo cells. The results of the present study demonstrated that JARID2 may serve a role in the progression of preeclampsia. The knockdown of JARID2 inhibited the viability and invasion of trophoblast cells in preeclampsia by suppressing the PI3K/Akt signaling pathway. Therefore, JARID2 may serve as a novel potential target for treating preeclampsia.


Long-term effects of Garcinia cambogia/Glucomannan on weight loss in people with obesity, PLIN4, FTO and Trp64Arg polymorphisms.
Overweight and obesity are considered major health problems that contribute to increase mortality and quality of life. Both conditions have a high prevalence across the world reaching epidemic numbers. Our aim was to evaluate the effects of the administration of Garcinia cambogia (GC) and Glucomannan (GNN) on long-term weight loss in people with overweight or obesity. Prospective, not-randomized controlled intervention trial was conducted. We treated 214 subjects with overweight or obesity with GC and GNN (500 mg twice a day, each) for 6 months evaluating weight, fat mass, visceral fat, basal metabolic rate, and lipid and glucose blood profiles comparing them with basal values. Some patients were carriers of polymorphisms PLIN4 -11482G > A-, fat mass and obesity-associated (FTO) -rs9939609 A/T- and β-adrenergic receptor 3 (ADRB3) -Trp64Arg. Treatment produced weight loss, reducing fat mass, visceral fat, lipid and blood glucose profiles while increasing basal metabolic rate. Results were independent of sex, age or suffering from hypertension, diabetes mellitus type 2 or dyslipidemia and were attenuated in carriers of PLIN4, FTO, Trp64Arg polymorphisms. Administration of GC and GNN reduce weight and improve lipid and glucose blood profiles in people with overweight or obesity, although the presence of polymorphisms PLIN4, FTO and ADRB3 might hinder in some degree these effects. ISRCTN78807585, 19 September 2017, retrospective study.


Clinical presentations of acute pulmonary embolism: A retrospective cohort study.
We aimed to investigate whether the unusual clinical presentation of pulmonary embolism (PE) varies by the type of provocation. In this retrospective cohort study, we examined the electronic health records (EHR) records of all patients diagnosed with PE (upon presentation or during hospitalization) presented to our tertiary hospital during 2014 to 2019. Inclusion criteria were the diagnosis of acute PE and age above 18 years. Excluded were all patients to whom complete EHR were not available. The primary outcome was the main presenting symptom, categorized by a multidisciplinary consensus expert committee as either typical or atypical of PE. Comorbidities, vital signs, medications and laboratory results on presentations were recorded. 591 patients were included in the final analysis. Dyspnea was significantly less common and hemoptysis and chest pain more common in the unprovoked PE group (35%, 5%, and 25%, respectively) compared with nonmalignant (42.6%, 0%, and 16.3%) and malignancy-associated (47.7%, 0.9%, and 8.2%) PE (Pv = 0.02, 0.002 and 0.001, respectively). No recorded symptoms were the third most common presentation overall, accounting for a significantly (Pv < 0.001) higher proportion of PE patients with malignancy (19%) whereas atypical presentation was the hallmark of patients with nonmalignant provokation (19.7%) (Pv = 0.005). Accounting for multiple potential confounders, the risk of atypical or asymptomatic presentation was higher with lower heart rates (RR = 0.974 95%C.I. [0.957-0.990]) and higher pulse oximetry saturation (RR = 1.114 95%CI [1.034-1.201]). The clinical presentation of PE varies with different types of provoking factors, with atypical presentation most common in nonmalignant provocation and asymptomatic presentation most prevalent in patients with underlying malignancy. Further studies are needed to determine the effect of said variance on long term clinical outcomes.


Risk factors for transmission of HIV from mother to child in Bangui.
the prevalence of human immunodeficiency virus infection (HIV) in pregnant women in Central Africa is 4.5%. Although a program to prevent mother-child transmission of HIV has been implemented throughout the country, the number of infected infants continues to increase. In this context, our study sought to determine the rate of mother-child transmission of HIV and to identify the main risk factors. this prospective cross-sectional study took place from November 1, 2014, to November 1, 2015, in all 16 maternal and child health centers in Bangui. The study population comprised the infants born to HIV-positive mothers and seen at these centers for HIV testing 6 weeks after birth. The mothers were interviewed and their files examined to identify the risk factors of HIV transmission to their babies. the study included 656 infants. The maternal-fetal transmission rate of HIV was 7%. The main risk factors were the absence of antiretroviral prophylaxis for mothers or children and rupture of the membranes more than 24 hours before labor. HIV transmission from mother to child is a public health problem in Bangui. Vigorous action, such as interventions with antiretroviral medication, good obstetric practices, and especially the implementation of the 2013 WHO recommendations, should certainly reduce the number of new infections among newborns in Bangui.


Determination of fibronectin and its degradation products in bronchoalveolar lavage fluid.
In order to estimate, in sarcoidosis, the extent of degradation and the availability of functionally active fibronectin (FN) in the alveolar space, native and total (= native + degraded) FN were determined in the bronchoalveolar lavage (BAL) fluid of 28 non-smoking patients with sarcoidosis, 12 healthy non-smokers and 24 asymptomatic smokers. The FN was measured in unconcentrated BAL fluid in the presence of the protease inhibitor aprotinin: total FN was assayed by a double-antibody sandwich enzyme-linked immunoadsorbent assay (ELISA) and native FN by a gelatin-antibody sandwich ELISA. Both in controls and in sarcoid patients, concentrations of native FN were lower than those of total FN, indicating that FN is degraded in the alveolar space. Compared to non-smoking controls, the sarcoid patients had significantly increased concentrations of both native and total FN (p less than 0.001 for both) in the BAL fluid. Native FN in percentage of total FN was similar in non-smoking sarcoids and controls, representing about 80% and 70%, respectively. In contrast, native FN corresponded to only 30% in the group of control smokers, indicating higher degradation in smokers. Thus, in smokers, less intact collagen-binding FN is available for forming an extracellular matrix, which may be necessary for repair. On the other hand, the elevation of FN, especially in its native form, in sarcoidosis implies that more of it may be available for repair but, if in excess, also for the build-up of fibrotic tissue.(ABSTRACT TRUNCATED AT 250 WORDS)


Unexpected Hazards with Dental High Speed Drill.
An expected accident can happen at any time during a routine practice in the dental office due to the types of instruments used. One of the instruments used in routine dental practice is a high speed drill and a bur. If Personal Protective Equipment (PPE) is not practiced at any time in the dental office, very serious injuries could easily happen to the clinician, staff or to the patient.


A Glu209Lys substitution in DRG1/TaACT7, which disturbs F-actin organization, reduces plant height and grain length in bread wheat.
Plant height and grain size are two important agronomic traits that are closely related to crop yield. Numerous dwarf and grain-shape mutants have been studied to identify genes that can be used to increase crop yield and improve breeding programs. In this study, we characterized a dominant mutant, dwarf and round grain 1 (drg1-D), in bread wheat (Triticum aestivum L.). drg1-D plants exhibit multiple phenotypic changes, including dwarfism, round grains, and insensitivity to brassinosteroids (BR). Cell structure observation in drg1-D mutant plants showed that the reduced organ size is due to irregular cell shape. Using map-based cloning and verification in transgenic plants, we found that a Glu209Lys substitution in the DRG1 protein is responsible for the irregular cell size and arrangement in the drg1-D mutant. DRG1/TaACT7 encodes an actin family protein that is essential for polymerization stability and microfilament (MF) formation. In addition, the BR response and vesicular transport were altered by the abnormal actin cytoskeleton in drg1-D mutant plants. Our study demonstrates that DRG1/TaACT7 plays an important role in wheat cell shape determination by modulating actin organization and intracellular material transport, which could in the longer term provide tools to better understand the polymerization of actin and its assembly into filaments and arrays.


Combination of Dichloroacetate and Atorvastatin Regulates Excessive Proliferation and Oxidative Stress in Pulmonary Arterial Hypertension Development via p38 Signaling.
Pulmonary arterial hypertension (PAH) is a lethal disease generally characterized by pulmonary artery remodeling. Mitochondrial metabolic disorders have been implicated as a critical regulator of excessively proliferative- and apoptosis-resistant phenotypes in pulmonary artery smooth muscle cells (PASMCs). Dichloroacetate (DCA) is an emerging drug that targets aerobic glycolysis in tumor cells. Atorvastatin (ATO) is widely used for hyperlipemia in various cardiovascular diseases. Considering that DCA and ATO regulate glucose and lipid metabolism, respectively, we hypothesized that the combination of DCA and ATO could be a potential treatment for PAH. A notable decrease in the right ventricular systolic pressure accompanied by reduced right heart hypertrophy was observed in the DCA/ATO combination treatment group compared with the monocrotaline treatment group. The DCA/ATO combination treatment alleviated vascular remodeling, thereby suppressing excessive PASMC proliferation and macrophage infiltration. In vitro, both DCA and ATO alone reduced PASMC viability by upregulating oxidative stress and lowering mitochondrial membrane potential. Surprisingly, when combined, DCA/ATO was able to decrease the levels of reactive oxygen species and cell apoptosis without compromising PASMC proliferation. Furthermore, suppression of the p38 pathway through the specific inhibitor SB203580 attenuated cell death and oxidative stress at a level consistent with that of DCA/ATO combination treatment. These observations suggested a complementary effect of DCA and ATO on rescuing PASMCs from a PAH phenotype through p38 activation via the regulation of mitochondrial-related cell death and oxidative stress. DCA in combination with ATO may represent a novel therapeutic strategy for PAH treatment.


Developments in nuclear cardiology: transition from single photon emission computed tomography to positron emission tomography-computed tomography.
Recent advances in positron emission tomography (PET) instrumentation have paralleled those of multichannel computed tomography (CT) for cardiac applications. Whereas multichannel CT angiography provides information on the presence and extent of anatomical luminal narrowing of epicardial coronary arteries, stress myocardial perfusion PET provides information on the downstream functional consequences of such anatomic lesions. With the advent of hybrid PET/CT systems, such complementary information of anatomy and physiology can be realized immediately at the same imaging session. By acquiring dynamic, gated myocardial perfusion data, PET studies provide insight into impairment of regional coronary blood flow reserve and microvascular endothelial dysfunction. This paper presents recent developments in PET detector materials, acquisition modes, combined PET/CT scanners, rubidium-82 (Rb-82) gated myocardial perfusion studies and analysis methods for absolute myocardial blood flow quantification.


Steric Effect of Antioxidant Diels-Alder-Type Adducts: A Comparison of Sanggenon C with Sanggenon D.
Sanggenons C and D are two Diels-Alder-type adducts from Chinese crude drug Sang-bai-pi. Structurally, both sanggenons construct stereoisomers. In the study, they were comparatively determined using four antioxidant assays, including ferric ion reducing antioxidant power (FRAP) assay, Cu2+-reducing assay, 1,1-diphenyl-2-picryl-hydrazl (DPPH•)-scavenging assay, and 2,2'-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid radical (ABTS•⁺)-scavenging assay. Their Fe2+-binding reactions were explored using UV-Vis spectra. Finally, their cytoprotective effects were evaluated using flow cytometry. In electron transfer (ET)-based FRAP and Cu2+-reducing assays, sanggenon D was found to have lower IC50 values than sanggenon C; however, in multi-pathway-based DPPH•-scavenging and ABTS•⁺-scavenging assays, sanggenon C possessed lower IC50 values than sanggenon D. UV-Vis spectra suggested that sanggenon C generated a bathochromic-shift (286 nm → 302 nm) and displayed stronger UV absorption than sanggenon D. In flow cytometry, sanggenon C and sanggenon D, respectively, exhibited 31.1% and 42.0% early apoptosis-percentages towards oxidative-stressed mesenchymal stem cells (MSCs). In conclusion, both sanggenons may undergo multiple pathways (e.g., ET and Fe2+-binding) to protect MSCs against oxidative stress. In the mere ET aspect, sanggenon D possesses a higher level than sanggenon C, while in multi-pathway-based radical-scavenging, Fe2+-binding, and cytoprotection aspects, sanggenon C is more active than sanggenon D. These discrepancies can conclusively be attributed to the steric effect.


Protein kinase C isoenzymes in airway smooth muscle.
The protein kinase C (PKC) isoenzymes expressed by bovine tracheal smooth muscle (BTSM) were identified at the protein and mRNA levels. Western immunoblot analyses reliably identified PKCalpha, PKCbetaI and PKCbetaII. In some experiments immunoreactive bands corresponding to PKCdelta, PKCepsilon and PKCTheta were also labelled, whereas the gamma, eta and zeta isoforms of PKC were never detected. Reverse transcriptase PCR of RNA extracted from BTSM using oligonucleotide primer pairs designed to recognize unique sequences in the PKC genes for which protein was absent or not reproducibly identified by immunoblotting, amplified cDNA fragments that corresponded to the predicted sizes of PKCdelta, PKCepsilon and PKCzeta, which was confirmed by Southern blotting. Anion-exchange chromatography of the soluble fraction of BTSM following homogenization in Ca2+-free buffer resolved two major peaks of activity. Using epsilon-peptide as the substrate, the first peak of activity was dependent upon Ca2+ and 4beta-PDBu (PDBu=phorbol 12, 13-dibutyrate), and represented a mixture of PKCs alpha, betaI and betaII. In contrast, the second peak of activity, which eluted at much higher ionic strength, also appeared to comprise a combination of conventional PKCs that were arbitrarily denoted PKCalpha', PKCbetaI' and PKCbetaII'. However, these novel enzymes were cofactor-independent and did not bind [3H]PDBu, but were equally sensitive to the PKC inhibitor GF 109203X compared with bona fide conventional PKCs, and migrated on SDS/polyacrylamide gels as 81 kDa polypeptides. Taken together, these data suggest that PKCs alpha', betaI' and betaII' represent modified, but not proteolysed, forms of their respective native enzymes that retain antibody immunoreactivity and sensitivity to PKC inhibitors, but have lost their sensitivity to Ca2+ and PDBu when epsilon-peptide is used as the substrate.


Befloxatone (Synthelabo).
Synthelabo is developing befloxatone, a long-acting selective and reversible monoamine oxidase A (MAO-A) inhibitor, for the treatment of depression, social phobias and panic disorders. Phase III trials have commenced in France for depression. In 1995, phase II trials began in Europe and the US for the treatment of depression, social phobias and panic disorders. The results of a dose-ranging study conducted in Europe in patients with major depressive disorders confirmed the product's safety. Befloxatone has a good safety profile, lacking sedative, convulsant, anticholinergic and cardiovascular effects--particularly important is its negligible tyramine effect. Results from phase I trials suggest that it will be suitable for once-daily oral dosage. It has an elimination half-life of approximately 11 h in man and is rapidly absorbed with a Tmax of around 2.5 h. In a trial of befloxatone in 12 healthy elderly volunteers, an oral dose of 10 mg was administered and psychomotor and cognitive function were assessed using objective tests such as critical flicker fusion (CFF), body sway and learning memory tasks amongst others and subjective tests such as the visual analog scale and a sleep questionnaire. Using these criteria, the 10 mg dose of befloxatone was shown not to produce any detrimental effects in skilled performance while a significant improvement in long term memory was shown. It appears to have no significant effects on skilled performance, memory or sleep patterns. [11C]-befloxatone was evaluated and found to be an excellent ligand for the study of MAO-A by positron emission tomography.


New developments in the immunology of asthma, with a focus on mechanisms and treatment.
Asthma is a chronic inflammatory disease of the airways involving the activation of many different cell types. The immunologic basis of asthma is widely accepted for allergic asthma, and the synthesis of IgE is dependent on cytokines. A process of healing and repair follows on from inflammation and this leads to a variable degree of remodelling of the airways. The treatment of asthma should take into account that the disease is associated with short-lived symptoms that can be prevented or reversed by bronchodilators, exacerbations that can be treated or reversed by anti-inflammatory drugs, and remodelling of the airways, for which no definite treatment has yet been validated.


Non-specific tumoral markers in hepatocellular carcinoma.
The number of biological tumoral markers used in the diagnosis and therapy monitoring of hepatocellular carcinoma has increased, but their separate use is limited as none of them is specific, being only tumour-associated (proteins). But when in abnormal amounts and used in combination, they are of great help in the diagnosis and therapy monitoring. A combination of alpha-fetoprotein (AFP) and alpha 1-antitrypsin (AAT) data raises the diagnostic accuracy in hepatocellular carcinoma from 43% obtained with AFP alone, to 90.5% and if the combination includes the carcinoembryonic antigen (CEA) data too the accuracy increases to 100%, still without strict specificity for hepatocellular carcinoma.


Neoplastic transformation of cultured mammalian cells by estrogens and estrogenlike chemicals.
Estrogens are clearly carcinogenic in humans and rodents but the mechanisms by which these hormones induce cancer are only partially understood. Stimulation of cell proliferation and gene expression by binding to the estrogen receptor is one important mechanism in hormonal carcinogenesis; however, estrogenicity is not sufficient to explain the carcinogenic activity of all estrogens because some estrogens are not carcinogenic. Estrogens are nonmutagenic in many assays but exhibit specific types of genotoxic activity under certain conditions. We have studied extensively the mechanisms by which estrogens induce neoplastic transformation in a model in vitro system and our findings are summarized in this review. 17beta-Estradiol (E2) and diethylstilbestrol (DES) and their metabolites induce morphological and neoplastic transformation of Syrian hamster embryo (SHE) cells that express no measurable levels of estrogen receptor. Treatment of the cells with E2 or DES fails to induce DNA damage, chromosome aberrations and gene mutations in SHE cells but results in numerical chromosome aberrations (aneuploidy) that could arise from microtubule disruption or disfunction of mitotic apparatus. Estrogen-induced genotoxicity is detected in cells following treatment with estrogen metabolites or following exogenous metabolic activation of estrogens. The estrogens induce DNA adduct formation that is detected by 32P-postlabeling. Both aneuploidy induction and DNA damage caused by DNA adduct formation correlate with the estrogen-induced cell transformation and may be important in hormonal carcinogenesis. We propose that multiple effects of estrogens acting together cause genetic alterations leading to cell transformation.


Reconstructing atomistic detail for coarse-grained models with resolution exchange.
Coarse-grained (CG) modeling has emerged as a promising tool to bridge the gap between the temporal and spatial scales of all-atom (AA) simulations and those of many important biological processes. Resolution exchange, a variant of the replica exchange method, combines the efficiency of CG simulation and the accuracy of AA simulation by swapping configurations between AA and CG simulations. The crucial step in a resolution exchange move is to rigorously reconstruct the high-resolution system from models at coarser resolutions. In this paper, configurational-bias Monte Carlo is adopted as a general method to rebuild the missing degrees of freedom rigorously for CG models and for the first time combined with resolution exchange. The new approach is demonstrated on an alkane and a peptide system. It is found that the efficiency of resolution exchange depends significantly on the quality of the CG model.


Generation of induced pluripotent stem cells, KCi002-A derived from a patient with Bardet-Biedl syndrome homozygous for the BBS10 variant c.271insT.
Bardet-Biedl syndrome (BBS) is genetically heterogeneous with at least 21 genes involved, and BBS10 encodes, together with BBS6 and BBS12, chaperonin-like proteins which are important for the assembly of the multiprotein complex, the BBSome encoded by other BBS genes. Here we describe the successful generation of an induced pluripotent stem cell (iPSC) line KCi002-A from a male with BBS, homozygous for the disease causing variant c.271insT, p.(Cys91fsX95) in BBS10. Resource table.


A day-hospital approach to treatment of pediatric complex regional pain syndrome: initial functional outcomes.
To examine clinical outcomes of an interdisciplinary day-hospital treatment program (comprised of physical, occupational, and cognitive-behavioral therapies with medical and nursing services) for pediatric complex regional pain syndrome (CRPS). The study is a longitudinal case series of consecutive patients treated in a day-hospital pediatric pain rehabilitation program. Participants were 56 children and adolescents with ages 8 to 18 years (median=14 y) with CRPS spectrum conditions who failed to progress sufficiently with a previous outpatient and/or inpatient treatments. Patients participated in daily physical therapy, occupational therapy, and psychological treatment and received nursing and medical care as necessary. The model places equal emphasis on physical and cognitive-behavioral approaches to pain management. Median duration of stay was 3 weeks. Outcome measures included assessments of physical, occupational, and psychological functioning at program admission, discharge, and at posttreatment follow-up at a median of 10 months after discharge. Scores at discharge and follow-up were compared with measures on admission by Wilcoxon tests, paired t tests, or analysis of variance as appropriate, with corrections for multiple comparisons. Outcomes demonstrate clinically and statistically significant improvements from admission to discharge in pain intensity (P<0.001), functional disability (P<0.001), subjective report of limb function (P<0.001), timed running (P<0.001), occupational performance (P<0.001), medication use (P<0.01), use of assistive devices (P<0.001), and emotional functioning (anxiety, P<0.001; depression, P<0.01). Functional gains were maintained or further improved at follow-up. A day-hospital interdisciplinary rehabilitation approach seems effective in reducing disability and improving physical and emotional functioning and occupational performance among children and adolescents with CRPSs that have failed to improve with outpatient treatment.


Are we any closer to treating liver fibrosis (and if no, why not)?
This review provides a personal view on anti-fibrosis therapy in the liver. The worst clinical consequence of liver fibrosis is the development of liver cirrhosis and portal hypertension. Etiology is a decisive factor which determines patterns of fibrous septa and subsequent vascular remodeling, which is essential for the development of portal hypertension. Removing or controlling the disease-causing agent, i.e. anti-viral treatment for hepatitis, is the essential first step for treating chronic liver diseases and can reverse fibrosis in some settings. However, removing etiology is not always sufficient to prevent fibrosis from progressing towards cirrhosis and portal hypertension. In liver diseases such as severe alcoholic hepatitis and massive parenchymal loss, the formation of vascular anastomoses between portal to central veins based on bridging fibrosis results in cirrhosis and portal hypertension. For these patients, anti-fibrotic treatment is crucial and urgent. Unfortunately, a lack of understanding how fibrosis contributes to vascular remodeling caused by and combined with a lack of suitable experimental models that recapitulate human liver diseases, has hampered the development of successful anti-fibrotic drugs for clinical use to date.


[Determination of five synthetic musks in perfume by headspace solid-phase microextraction and gas chromatography-mass spectrometry].
A method for headspace solid-phase microextraction (HS-SPME), followed by gas chromatography-mass spectrometry (GC-MS) analysis was established for the determination of five commonly used synthetic musks in perfume. Two polycyclic musks (celestolide and tonalide) and three nitro musks (musk ambrette, musk xylene and musk ketone) were used as analytes in the optimization of the analytical method. Six parameters, such as the extraction temperature, equilibrium time, extraction time, desorption time, injector temperature and solution of salting out, were optimized by exposing the 65 microm polydimethylsiloxane-divinyl-benzene (PDMS-DVB) fiber to the headspace of magnetically stirred (600 r/min) sample. According to the results of the optimization experiments, the following conclusion can be drawn: The water-diluted sample in a 10 mL headspace-vial was efficiently extracted for 20 min after the system was equilibrated for 3 min at 60 degrees C. After extraction, the fiber was immediately inserted into the GC injector and desorbed at 250 degrees C for 3 min. The spiked recoveries were in the range of 82.0% - 103.3% and the relative standard deviations (RSDs) were between 1.8% and 9.4%. Meanwhile, the limits of detection (LODs) ranged from 0.6 ng/g to 2.1 ng/g. This method is characterized by rapidity, high sensitivity, good linearity and repeatability for all the target compounds. It is applicable to the analysis of synthetic musks in perfumes.


High-throughput transcriptome sequencing and comparative analysis of Escherichia coli and Schizosaccharomyces pombe in respiratory and fermentative growth.
In spite of increased complexity in eukaryotes compared to prokaryotes, several basic metabolic and regulatory processes are conserved. Here we explored analogies in the eubacteria Escherichia coli and the unicellular fission yeast Schizosaccharomyces pombe transcriptomes under two carbon sources: 2% glucose; or a mix of 2% glycerol and 0.2% sodium acetate using the same growth media and growth phase. Overall, twelve RNA-seq libraries were constructed. A total of 593 and 860 genes were detected as differentially expressed for E. coli and S. pombe, respectively, with a log2 of the Fold Change ≥ 1 and False Discovery Rate ≤ 0.05. In aerobic glycolysis, most of the expressed genes were associated with cell proliferation in both organisms, including amino acid metabolism and glycolysis. In contrast in glycerol/acetate condition, genes related to flagellar assembly and membrane proteins were differentially expressed such as the general transcription factors fliA, flhD, flhC, and flagellum assembly genes were detected in E. coli, whereas in S. pombe genes for hexose transporters, integral membrane proteins, galactose metabolism, and ncRNAs related to cellular stress were overexpressed. In general, our study shows that a conserved "foraging behavior" response is observed in these eukaryotic and eubacterial organisms in gluconeogenic carbon sources.


Highly selective directed assembly of functional actomyosin on Au surfaces.
The capability of assembling biomotors onto specific locations of solid substrates is a key for development of biomotor-based nanomechanical systems. We developed a method to direct the assembly of the heavy meromyosin fragment from rabbit skeletal muscle myosin onto specific locations of Au substrates utilizing surface molecular patterns. In this strategy, chemically directed patterns of streptavidin were achieved to direct highly specific assembly of biotinylated heavy meromyosin on the substrates--a strategy applicable for patterning a variety of biotinylated molecules--while BSA was utilized to avoid nonspecific adsorption. In vitro motility assays of filament sliding were used to confirm functionality of assembled actomyosin.


Anaerobic α-amylase production and secretion with fumarate as the final electron acceptor in Saccharomyces cerevisiae.
In this study, we focus on production of heterologous α-amylase in the yeast Saccharomyces cerevisiae under anaerobic conditions. We compare the metabolic fluxes and transcriptional regulation under aerobic and anaerobic conditions, with the objective of identifying the final electron acceptor for protein folding under anaerobic conditions. We find that yeast produces more amylase under anaerobic conditions than under aerobic conditions, and we propose a model for electron transfer under anaerobic conditions. According to our model, during protein folding the electrons from the endoplasmic reticulum are transferred to fumarate as the final electron acceptor. This model is supported by findings that the addition of fumarate under anaerobic (but not aerobic) conditions improves cell growth, specifically in the α-amylase-producing strain, in which it is not used as a carbon source. Our results provide a model for the molecular mechanism of anaerobic protein secretion using fumarate as the final electron acceptor, which may allow for further engineering of yeast for improved protein secretion under anaerobic growth conditions.


Piezoelectric quartz crystal microbalance sensor for trace aqueous cyanide ion determination.
Using selective reaction chemistry, our present research has developed an online, real-time sensor capable of monitoring toxic cyanide at both drinking water standard and environmental regulatory concentrations. Through the use of a flow cell, aqueous samples containing cyanide are reacted with a gold electrode of a piezoelectric crystal to indirectly sense cyanide concentration by the dissolution of metallic gold. The quartz crystal is an AT-cut wafer sandwiched between two neoprene O-rings within the liquid flow cell. The presence of cyanide in solution results in the selective formation of a soluble dicyano-gold complex according to the Elsner reaction: 4Au + 8CN- + 2H2O + O2 <=> 4Au(CN)2- + 4OH-. The resulting loss of gold from the electrode is detected by the piezoelectric crystal as a resonant frequency change. Since free cyanide is a weak acid (pKa = 9.3), available protons compete for cyanide ligands. Therefore, increased sample pH provides higher sensitivity. The detection limits at pH 12 are 16.1 and 2.7 ppb for analysis times of 10 min and 1 h, respectively. The incorporation of the flow cell improves both analyte sensitivity and instrument precision, with an average signal intensity drift of only 5% over a 2-h analysis. The calibrations show excellent linearity over a variety of cyanide concentrations ranging from low ppb to hundreds of ppm. This detection method offers the advantage of selectively detecting cyanides posing a biohazard while avoiding detection of stable metal cyanides. This aspect of the system is based on competitive exchange of available metals and gold with cyanide ligands. Stable metal cyanide complexes possess a higher formation constant than cyanoaurate. This detection system has been configured into a flow injection analysis array for simple adaptation to automation. Anions commonly found in natural waters have been examined for interference effects. Additionally, the sensor is free from interference by aqueous cyanide analogues including thiocyanate. The developed detection system provides rapid cyanide determinations with little sample preparation or instrument supervision.


Neutrophil-Lymphocyte Ratio in Different Stages of Breast Cancer.
Despite many advances in the treatment of breast cancer, it is still the second most common cause of death in women in the United States. It has been shown that inflammation plays a major role in the treatment of these cancers and inflammatory factors enhance tumor growth, invasion, metastasis, and vascularization. In this study, we would like to analyze peripheral blood neutrophil-lymphocyte ratio (NLR) in breast cancer patients and its correlation with disease staging. This cross-sectional analytic study was conducted in Imam Hospital, affiliated with Tehran University of Medical Sciences; a total of 195 female patients with breast cancer met the inclusion criteria. All of the patients had a complete blood count with leukocyte differential performed before chemotherapy. Medical records including pathology reports were also available. Data for all patients were collected prior to any surgical intervention. Exclusion criteria included clinical evidence of active infection, presence of hematological disorders, acute as well as chronic inflammatory or autoimmune diseases, or prior steroid therapy. Higher platelet count was significantly associated with the higher stage. The stage was not associated with the hemoglobin level. There was no association between the tumor size and age of patients with NLR. There was a significant relationship between NLR and IDC. There was a significant relationship between IDC and NLRs of less than 8.1 and greater than 3.3. There was a significant relationship between NLR and vascular invasion. There was no association between NLR and estrogen receptor and HER2. There was no significant relationship between the PLR and the cancer stage. In this study, NLR showed a significant relation with the disease staging. As the NLR increases the stage increases as well. Therefore, this ratio may be helpful in the preoperative evaluation of patients with breast cancer.


Antioxidant Effect of Fructus Ligustri Lucidi Aqueous Extract in Ovariectomized Rats Is Mediated through Nox4-ROS-NF-κB Pathway.
Purpose: This study is designed to explore whether Fructus ligustri lucidi (FLL) exhibits antioxidant effect in ovariectomized (OVX) rats, and to identify the signaling pathway involved in this process. Methods: OVX rats were treated with FLL aqueous extract (3.5 g/kg) for 12 weeks. Serum, uteri, and tibias were harvested from the rats and the levels of total antioxidant capacity (TAC), nitric oxide (NO), malondialdehyde (MDA), 8-hydroxy-desoxyguanosine (8-OHdG), and superoxide dismutase (SOD) were determined. Changes in the levels of NF-κB-p65, phosphorylation of NF-κB-p65 (NF-κB-pp65), NF-κB inhibitor alpha (IκBα), phosphorylation of IκBα (p-IκBα), and NADPH oxidase 4 (Nox4) in uteri and tibias were determined by western blot, immunofluorescent and immunohistochemical analysis, respectively. In addition, the expression of cytochrome C (Cyto-C) and B-cell lymphoma-2 (Bcl-2) were determined in the tibias of rats. Histopathological changes in the bones were evaluated by hematoxylin-eosin staining. Bone mineral density (BMD) was determined in rat femurs by dual X-ray absorptiometry. Results: Treatment of OVX rats with FLL aqueous extract improved redox homeostasis by increasing the levels of TAC and NO as well as decreasing the levels of MDA and 8-OHdG in serum, tibias, and uteri. Further, FLL extract also downregulated the expression of Nox4, NF-κB-p65, NF-κB-pp65, and p-IκBα in the uteri and tibias. Furthermore, administration of FLL-OVX rats increased Bcl-2 expression and prevented cytoplasmic release of mitochondrial Cyto-C in the tibias. In addition, FLL treatment also improved bone microstructure and increased cortical bone thickness as well as increased BMD values in the femurs of OVX rats. Conclusions: FLL treatment may suppress oxidative stress response in OVX rats via regulating the Nox4/ROS/NF-κB signaling pathway. These results suggest the potential of using FLL as a natural antioxidant agent in preventing the development of osteoporosis.


Efficient perovskite/organic integrated solar cells with extended photoresponse to 930 nm and enhanced near-infrared external quantum efficiency of over 50.
Enhancing the light-harvesting activity is an effective way to improve the power conversion efficiency of solar cells. Although rapid enhancement in the PCE up to a value of 22.1% has been achieved for perovskite solar cells, only part of the sunlight, i.e., with wavelengths below 800-850 nm is utilized due to the limited bandgap of the perovskite materials, resulting in most of the near infrared light being wasted. To broaden the photoresponse of perovskite solar cells, we demonstrate an efficient perovskite/organic integrated solar cell containing both CH3NH3PbI3 perovskite and PBDTTT-E-T:IEICO organic photoactive layers. By integrating a low band gap PBDTTT-E-T:IEICO active layer on a perovskite layer, the maximum wavelength for light harvesting of the ISC increased to 930 nm, sharply increasing the utilization of near infrared radiation. In addition, the external quantum efficiency of the integrated device exceeded 50% in the near infrared range. The MAPbI3/PBDTTT-E-T:IEICO ISCs show an enhanced short-circuit current density of over 24 mA cm-2, which is the highest existing value among perovskite/organic integrated solar cells and much higher than the traditional MAPbI3 based perovskite solar cells. The results reveal that a perovskite/organic integrated structure is a promising strategy to extend and enhance sunlight utilization for perovskite solar cells.


[Effectiveness of imipenem/cilastatin (Tienam, MSD) in treating complicated infections in urology].
Complicated urinary infections tend to eventuate in severe pyoseptic complications--bacteriuria, sepsis. The search for methods of fighting agents of urinary infections goes in the direction of perfection of already existing methods and in the direction of design of novel antibacterial drugs. In the middle 1980s the first carbapenem drug-imipenem--was proposed for urological clinical practice. Mechanism of its action as that of the other beta-lactam antibiotics consists in impairment of synthesis of bacterial cell wall as a result of the drug penetration through the surface membrane and irreversible binding with penicillin-binding proteins. Imipenem is active against most gram-positive and gram-negative aerobic and anaerobic microorganisms which cause severe urological infections. The article presents the results of treatment of 45 patients with severe urological infections with multiple resistance of the causing agent and failure of previous treatment. Imipenem was given in a daily dose 1.5-2.0 g. Sometimes a stepwise regimen was used: 500 mg 4 times a day intravenously for the first 3-4 days, then 500 mg twice a day intramuscularly for the following 3-4 days. In detection of highly sensitive bacteria (E. coli, Proteus mirabilis) daily doses were reduced to 1 g. In long standing infection caused by Pseudomonas aeruginosa imipenem was combined with amicacin. In high surgical risk of postoperative period imipenem was given prior to surgery and continued after it for 5 to 14 days. Good therapeutic results were achieved: clinical effect reached 95.5%, antibacterial efficiency was 87.8%. Thus, imipenem is antibiotic of the first line in empirical therapy of severe bacterial infections in urology as it has a wide spectrum of antibacterial action. We believe that this drug should not be left as a reserve but used for a starting empirical therapy of severe infections in urological hospital.


Development and validation of the Patient-Physician Relationship Scale among patients with irritable bowel syndrome.
An effective patient-physician relationship (PPR) is essential to the care of patients with irritable bowel syndrome (IBS). We sought to develop and validate an IBS-specific instrument to measure expectations of the PPR. We conducted structured focus groups about PPRs with 12 patients with IBS. Qualitative analysis was used to generate a questionnaire (the Patient-Physician Relationship Scale [PPRS]), which was modified with input from content experts and usability testing. For validation, we administered it online to US adults with IBS. Participants also completed the Functional Bowel Disorder Severity Index, the Rome III Adult Functional gastrointestinal (GI) Disorder Criteria Questionnaire, and modified versions of the Communication Assessment Tool (CAT-15) and Patient-Doctor Relationship Questionnaire (PDRQ-9). We performed principal components factor analysis for the PPRS. The PPRS contained 32 questions with responses on a 7-item Likert scale. Themes included interpersonal features, clinical care expectations, and aspects of communication. One thousand and fifty-four eligible individuals completed the survey (88% completion rate). Most participants were middle aged (mean 48 years, SD 16.3), white (90%), and female (86%). Factor analysis showed only one relevant factor, relating to quality of PPR. The final scale ranged from possible-96 to +96 (mean 62.0, SD 37.6). It correlated moderately with the CAT-15 (r=.40, P<.001) and PDRQ-9 (r=.30, P<.001), establishing concurrent validity. We describe the development and validation of the first questionnaire for use in measuring patient expectations of the PPR, which can be used for future outcomes studies and training physicians.


Aggressiveness study on Sclerotinia isolates from red clover crops.
Sclerotinia trifoliorum Erikks. causes clover rot (clover cancer, Sclerotinia crown and root rot) in red clover crops (Trifolium pratense L.), an important disease in Europe. Little is known about the aggressiveness of Scierotinia isolates and aggressiveness studies were never conducted on a European scale. In this study we compared the aggressiveness of 30 Sclerotinia isolates isolated from red clover crops in 25 locations in 12 European countries using a plant-based bio-test. Plants from 6 red clover cultivars with different resistance levels were spray inoculated at the age of 12 weeks with 1 to 1.5 ml mycelium fragment suspension per plant. After 10 days incubation, plants were scored on a scale from 1 (healthy plant) to 5 (dead plant) and the disease index was calculated. The experiment was repeated 3 times and all repetitions were highly correlated. Average disease indices ranged from 52.6% to 82.7%. Significant differences were detected between isolates and between cultivars, but there was no isolate--cultivar interaction. Based on these results, the most aggressive isolates can be selected for resistance breeding. Future work should investigate whether the differences in aggressiveness are due to a higher growth speed or due to a higher secretion of cell-wall degrading components and pathogenicity factors.


CRISPR-Cas mediated genome engineering of cyanobacteria.
Over the past decade, several cyanobacterial strains have emerged as exciting model systems for unraveling important biological process like photosynthesis and nitrogen fixation. In parallel, novel strains are being developed as platforms for production of various value-added products. To meet either of these goals, synthetic biology tool development has been prioritized, and among many such tools, CRISPR-mediated genome editing tools distinctly hold the potential to revolutionize cyanobacterial research. This chapter reviews our current understanding of the existing and emerging CRISPR-based technologies and their potential application for advanced genome editing in cyanobacterial strains of interest. CRISPR-based tools have gained traction in cyanobacterial research for their ability to target the polyploid genomes in these organisms and generate fully segregated mutants in a remarkably short time. We discuss the native cyanobacterial CRISPR system and the promise they hold for use as precision tools for cyanobacterial genome editing. We elaborate the methodologies for the development of CRISPR-based markerless mutants in cyanobacteria as well as discuss strategies for large scale, regulated genome silencing with CRISPRi. We also highlight some of the emerging CRISPR tools that have shown promise in other prokaryotic and eukaryotic systems but are yet to be adapted for cyanobacterial research.


Cloning and molecular characterization of the cDNAs encoding the variable regions of an anti-CD20 monoclonal antibody.
CD20-based targeting of B-cells in hematologic malignancies and autoimmune disorders is associated with outstanding clinical outcomes. Isolation and characterization of VH and VL cDNAs encoding the variable regions of the heavy and light chains of monoclonal antibodies (MAb) is necessary to produce next generation MAbs and their derivatives such as bispecific antibodies (bsAb) and single-chain variable fragments (scFv). This study was aimed at cloning and characterization of the VH and VL cDNAs from a hybridoma cell line producing an anti-CD20 MAb. VH and VL fragments were amplified, cloned and characterized. Furthermore, amino acid sequences of VH, VL and corresponding complementarity-determining regions (CDR) were determined and compared with those of four approved MAbs including Rituximab (RTX), Ibritumomab tiuxetan, Ofatumumab and GA101. The cloned VH and VL cDNAs were found to be functional and follow a consensus pattern. Amino acid sequences corresponding to the VH and VL fragments also indicated noticeable homologies to those of RTX and Ibritumomab. Furthermore, amino acid sequences of the relating CDRs had remarkable similarities to their counterparts in RTX and Ibritumomab. Successful recovery of VH and VL fragments encourages the development of novel CD20 targeting bsAbs, scFvs, antibody conjugates and T-cells armed with chimeric antigen receptors.


Conceptualizing physical activity behavior of older Korean-Americans: an integration of Korean culture and social cognitive theory.
People can live longer and healthier lives by engaging in physical activity (PA). The purpose of this article is to assess the social cognitive theory (SCT) in relation to its relevance to produce cultural-specific directions for gerontological nursing practice in order to guide the design of PA interventions for Korean-American elders. SCT is compared to the Korean cultural, social, and health belief system and is analyzed and evaluated based on 3 criteria: assumptions of the theory, completeness and consistency, and essence of nursing. Within the Korean culture, as presumed in the SCT and the nursing paradigm, health-promoting behavior, such as PA, is conceptualized as the desire for a higher level of health rather than a fear of disease as is proposed by other health behavior theories. SCT with the integration of Korean culture recognizes cultural, developmental, societal, and other external constraints that may help in formulating interventions and better understanding of the limits faced by older Korean-Americans (OKAs) in their pursuit of routine PA.


The psychometrics and validity of the Junior Temperament and Character Inventory in Portuguese adolescents.
Personality traits related to persistence and self-regulation of long-term goals can predict academic performance as well or better than measures of intelligence. The 5-factor model has been suggested to outperform some other personality tests in predicting academic performance, but it has not been compared to Cloninger's psychobiological model for this purpose. The aims of this study were, first, to evaluate the psychometric properties of the Junior Temperament and Character Inventory (JTCI) in adolescents in Portugal, and second, to evaluate the comparative validity of age-appropriate versions of Cloninger's 7-factor psychobiological model, Costa and McCrae's five-factor NEO-Personality Inventory-Revised, and Cattell's 16-personality-factor inventory in predicting academic achievement. All dimensions of the Portuguese JTCI had moderate to strong internal consistency. The Cattell's sixteen-personality-factor and NEO inventories provided strong construct validity for the JTCI in students younger than 17 years and for the revised adult version (TCI-Revised) in those 17 years and older. High TCI Persistence predicted school grades regardless of age as much or more than intelligence. High TCI Harm Avoidance, high Self-Transcendence, and low TCI Novelty Seeking were additional predictors in students older than 17. The psychobiological model, as measured by the JTCI and TCI-Revised, performed as well or better than other measures of personality or intelligence in predicting academic achievement.


Service quality and maturity of health care organizations through the lens of Complexity Leadership Theory.
This research focuses on Complexity Leadership Theory and the relationship between leadership-examined through the lens of Complexity Leadership Theory-and organizational maturity as an indicator of the performance of health organizations. The research adopts a perspective that conceptualizes organizations as complex adaptive systems and draws upon a survey of opinion of 189 managers working in Serbian health organizations. As the results indicate a dependency between functions of leadership and levels of the maturity of health organizations, we propose a model that connects the two. The study broadens our understanding of the implications of complexity thinking and its reflection on leadership functions and overall organizational performance. The correlations between leadership functions and maturity could have practical applications in policy processing, thus improving the quality of outcomes and the overall level of service quality.


Renal track creation for percutaneous nephrolithotomy: the history and relevance of single stage dilation.
Percutaneous nephrolithotomy (PCNL) was described in the 1980s and revolutionized the treatment of stone disease. The crucial component to this surgery is satisfactory track creation. We examine how the development and production in the 1980s of a single stage dilator (SSD) subsequently modified for pediatric PCNL has become the ideal access tool for mini percutaneous nephrolithotomy (mPCNL) today. The conception, production, scientific and clinical development of the original SSD is described. The pitfalls of track dilation in general according to method of dilation are also discussed and outlined. This study provides evidence clarifying commonly held misconceptions about the origin of SSD which is the mainstay of the mPCNL technique. Percutaneous renal surgery continues to evolve. In less than 40 years stone surgery has transformed from a morbid open operation to a number of minimally invasive, routine techniques. The SSD has been an innovation that has played a crucial role in this change.


Observation of vortex nucleation in a rotating two-dimensional lattice of Bose-Einstein condensates.
We report the observation of vortex nucleation in a rotating optical lattice. A 87Rb Bose-Einstein condensate was loaded into a static two-dimensional lattice and the rotation frequency of the lattice was then increased from zero. We studied how vortex nucleation depended on optical lattice depth and rotation frequency. For deep lattices above the chemical potential of the condensate we observed a linear dependence of the number of vortices created with the rotation frequency, even below the thermodynamic critical frequency required for vortex nucleation. At these lattice depths the system formed an array of Josephson-coupled condensates. The effective magnetic field produced by rotation introduced characteristic relative phases between neighboring condensates, such that vortices were observed upon ramping down the lattice depth and recombining the condensates.


Spontaneous Regression of Annular Basal Cell Carcinoma: A Case Report.
We herein present a case of annular basal cell carcinoma (BCC) with spontaneous regression on the right temporal region of a 56-year-old Japanese male. Histopathological examination revealed that the central section had no tumor cells. The sweat glands, follicles, and other cutaneous appendages were also absent. This pattern of spontaneous regression is quite uncommon, and understanding the histopathology may be important for future approaches to BCC.


Effects of repeated exposure on liking for a reduced-energy-dense food.
Reduced-energy-dense diet foods are often formulated to match the sensory characteristics of their regular-energy-dense counterparts. However, the extent to which attitudes toward a reduced-energy-dense food remain constant, even after repeated ingestion, remains to be explored systematically. The objective was to determine whether liking, "expected satiety," and "expected satiation" change after repeated exposure to a familiar food that has been reduced in energy density. Expected satiety and expected satiation refer to the extent to which foods are expected to stave off hunger and to deliver "fullness," respectively, when compared on a calorie-for-calorie basis. Participants (n = 36) consumed either reduced-energy-dense (374 kcal) or standard-energy-dense (567 kcal) spaghetti Bolognese for lunch over 5 test sessions. During each test session, liking for the spaghetti Bolognese was assessed, together with measures of expected satiety and expected satiation. Participants in the reduced-energy-dense condition reported a decrease in liking for the spaghetti Bolognese over the test sessions ( approximately 30%), whereas liking in the standard condition remained constant [condition (reduced/standard) x session (1-5) interaction, P < 0.008]. By contrast, both expected satiation and expected satiety remained similar across conditions and test sessions. Over time, the pleasantness of a reformulated low-energy-dense food can decrease, and this may undermine its efficacy as a weight-loss product. It remains to be determined whether a longer period of "flavor-nutrient learning" is needed for shifts in expected satiety and expected satiation to be observed.


Strongly Coupled Metal Oxide/Reassembled Carbon Nitride/Co-Pi Heterostructures for Efficient Photoelectrochemical Water Splitting.
The photoelectrochemical application of carbon nitride is extremely exciting because of the meta-free components, low cost, nontoxicity, and appropriate band positions. To construct carbon nitride-based heterostructures, a conventional ultrasonic exfoliation method is usually used to fabricate dispersion of ultrathin nanosheets. However, the outstretched structure and the poor dispersity inevitably result in the poor interfacial contact between different materials. To solve this problem, hydrolyzed carbon nitride suspension was used as a homogeneous precursor for the fabrication of composite photoanodes. The in situ reassembly of one-dimensional nanofibers resulted in the formation of uniform and ultrathin carbon nitride nanoarchitectures on the surface of Fe2O3 nanorod arrays. Because of the strongly coupled interfaces and the deposition of Co-Pi water oxidation cocatalysts, the as-synthesized heterostructured photoanodes exhibited three-fold increased photocurrent density and good stability, compared to pristine Fe2O3. The significantly improved photoactivity of the Fe2O3/reassembled carbon nitride/Co-Pi heterostructures was ascribed to the decreased interfacial conductivity and facilitated charge separation. This material designing strategy was further used to construct TiO2/carbon nitride, ZnO/carbon nitride, and WO3/carbon nitride heterostructures. The incorporation of hydrolyzed carbon nitride could remarkably enhance the photoelectrochemical performance of these metal oxide photoanodes. Thus, this work provides a new paradigm for designing carbon nitride-based composite nanostructures for efficient and stable solar fuel production.


Biological activities of extracts from sumac (Rhus spp.): a review.
Sumac is the common name for a genus (Rhus) that contains over 250 individual species of flowering plants in the family Anacardiaceae. These plants are found in temperate and tropical regions worldwide, often grow in areas of marginal agricultural capacity, and have a long history of use by indigenous people for medicinal and other uses. The research efforts on sumac extracts to date indicate a promising potential for this plant family to provide renewable bioproducts with the following reported desirable bioactivities: antifibrogenic, antifungal, antiinflammatory, antimalarial, antimicrobial, antimutagenic, antioxidant, antithrombin, antitumorigenic, antiviral, cytotoxic, hypoglycaemic, and leukopenic. As well, the bioactive components can be extracted from the plant material using environmentally benign solvents that allow for both food and industrial end-uses. The favorable worldwide distribution of sumac also suggests that desirable bioproducts may be obtained at the source, with minimal transportation requirements from the source through processing to the end consumer. However, previous work has focussed in just a few members of this large plant family. In addition, not all of the species studied to date have been fully characterized for potential bioactive components and bioactivities. Thus, there remains a significant research gap spanning the range from lead chemical discovery through process development and optimization in order to better understand the full potential of the Rhus genus as part of global green technology based on bioproducts and bioprocesses research programs.


Development of an open-source web-based intervention for Brazilian smokers - Viva sem Tabaco.
Web-based interventions for smoking cessation available in Portuguese do not adhere to evidence-based treatment guidelines. Besides, all existing web-based interventions are built on proprietary platforms that developing countries often cannot afford. We aimed to describe the development of "Viva sem Tabaco", an open-source web-based intervention. The development of the intervention included the selection of content from evidence-based guidelines for smoking cessation, the design of the first layout, conduction of 2 focus groups to identify potential features, refinement of the layout based on focus groups and correction of content based on feedback provided by specialists on smoking cessation. At the end, we released the source-code and intervention on the Internet and translated it into Spanish and English. The intervention developed fills gaps in the information available in Portuguese and the lack of open-source interventions for smoking cessation. The open-source licensing format and its translation system may help researchers from different countries deploying evidence-based interventions for smoking cessation.


Near-isogenic lines for resistance to brown planthopper with the genetic background of Indica Group elite rice (Oryza sativa L.) variety 'IR64'.
The brown planthopper (BPH), Nilaparvata lugens Stål, is an insect pest that severely damages rice (Oryza sativa L.) in Asia, causing huge yield loss. Use of resistant variety is a cost-effective and eco-friendly strategy for maintaining BPH populations below the economic injury level. However, current BPH populations have been changed to virulence against resistant varieties. In this study, to estimate effective combinations among eight BPH resistance genes (BPH32, BPH17-ptb, BPH20, BPH17, BPH3, BPH25, BPH26 and qBPH6), eight near-isogenic lines with the genetic background of an Indica Group rice variety 'IR64' (IR64-NIL) were developed using marker-assisted selection. The genome recoveries of these NILs ranged from 89.3% to 98.8% and agronomic traits of them were similar to those of 'IR64'. In modified seed box screening test, resistance level of IR64-NILs was higher than that of 'IR64'. In antibiosis test, high adult mortalities of BPH (from 56.0% to 97.0%) were observed among NILs, in comparison with that of 'IR64'. Among IR64-NILs, the line carrying BPH17 showed the highest resistance level at all tests. Thus, these IR64-NILs with multiple BPH resistance genes could be valuable breeding lines for enhancing resistance levels by gene pyramiding and multiline variety.


Drug eluting stents versus PTA with GP IIb/IIIa blockade below the knee in patients with current ulcers--The BELOW Study.
The study was designed as a feasibility trial to evaluate the use of GP IIb/IIIa blockade in connection with drug eluting stents, bare stents and PTA only. Sixty patients with current ulcers were randomly assigned to receive abciximab plus a sirolimus coated stent (N.=14), abciximab plus a bare stent (N.=16), abciximab plus PTA (N.=14) and PTA alone (N.=19). Angiographic control was performed at two and six months. Recanalization was successful in all cases. Two month restenosis rate was 9%, 45.5%, 67% and 46%. At six month follow-up restenosis rate was 9%, 67%, 75% and 58%, respectively; 14% of all patients had major amputations within six months. Adjunctive administration of abciximab during peripheral arterial intervention below the knee was found to be safe. Sirolimus coated stent administration was followed by a higher patency rate.


Anisotropic conduction properties in canine atria analyzed by high-resolution optical mapping: preferential direction of conduction block changes from longitudinal to transverse with increasing age.
Anisotropic conduction properties may provide a substrate for reentrant arrhythmias. We investigated the age-dependent changes of structural and functional anisotropy in isolated right atria from infant (1 to 2 months), young (6 to 12 months), and old (6 to 10 years) dogs. The histology of the mapped atrial tissues (a small subepicardial area, 2.8x4.2 mm) was characterized by an age-dependent increase of myofiber width and fat cell infiltration between myofibers. Cx43 was distributed homogeneously over the entire cell surface in infant dogs, whereas it progressively polarized to the cell termini with increasing age. The activation sequences were analyzed by high-resolution optical mapping using a voltage-sensitive dye. Activation fronts from the pacing site proceeded more rapidly along fiber orientation (longitudinal) than across it (transverse). Infant dogs showed "elliptical" isochrones with a smooth transition between longitudinal and transverse propagation, whereas old dogs had a "square" pattern with a sharp transition. Conduction block occurred predominantly during longitudinal propagation in infant dogs but during transverse propagation in old dogs. The shape of the wave front and the degree of lateral uncoupling seemed to decide the preferential direction of block. A zigzag activation causing an extremely slow transverse conduction was observed only in old dogs. Along with the age-dependent structural anisotropy, the preferential direction of block changed from longitudinal to transverse in association with a change in the wave front configuration. A zigzag propagation based on lateral uncoupling would predispose the elderly to multiple reentry and a higher incidence of atrial fibrillation.


Clinical outcomes of revision lumbar spinal surgery: 124 patients with a minimum of two years of follow-up.
Pertinent literature on revision lumbar spinal surgery has revealed a wide variation in success rates, ranging from 12% to 82%. In addition, a solid consensus has not yet been reached on its positive factors. We retrospectively reviewed 124 consecutive patients who underwent revision lumbar spinal surgery and investigated the factors that affected the outcomes of their surgery. Revision lumbar spinal surgery was performed in 124 patients from January 1992 to December 1996, with an average follow-up of 37.6 months (range, 24-89 months). The various factors analyzed included age, gender, previous diagnosis, number of previous operations, period of pain-free interval, neurologic deficit, operative procedure, and fusion status. This analysis revealed the effect of each factor on the overall results. Radiographs were obtained, and patients were assessed during the final follow-up or by questionnaire. The success rate of revision lumbar spine surgery in this study was 83.9%. Successful outcomes were significantly associated with the spinal procedure with fusion and with union of the spinal fusion. Patients with defined mechanical instability had better results than did those with stenosis only. In addition, the complication rate for repeated lumbar spinal surgery was 9.6% and major complications attributed to poor results. This study reveals a high success rate of revision spinal surgery. We recommend performing spinal fusion, and achievement of solid fusion in repeated low back surgery is invaluable for patients with spinal instability. Targeting the specific pathology of failed back surgery syndrome is crucial in attaining satisfactory results with revision lumbar spinal surgery.


Agreement between PiCCO pulse-contour analysis, pulmonal artery thermodilution and transthoracic thermodilution during off-pump coronary artery by-pass surgery.
Haemodynamic instability during off-pump coronary artery bypass surgery (OPCAB) may appear rapidly, and continuous monitoring of the cardiac index (CI) during the procedure is advisable. With the PiCCO monitor, CI can be measured continuously and almost real time with pulse-contour analysis and intermittently with transthoracic thermodilution. The agreement between pulmonal artery thermodilution CI (Tpa), transthoracic thermodilution CI (Tpc) and pulse-contour CI (PCCI) during OPCAB surgery has not been evaluated sufficiently. In 30 patients scheduled for OPCAB surgery, a pulmonary artery catheter and a PiCCO catheter were inserted. At different time points during surgery, Tpa, Tpc and PCCI were compared. Measurements were performed after induction of anesthesia (T1), after pericardiothomy (T2), after grafting on the anterior (T3), posterior (T4) and lateral (T5) walls and after chest closure (T6). The PCCI was recalibrated at time point T2-T6. Mean difference and the limits of agreements (percentage error) between Tpa and Tpc were: -0.14 +/- 0.60 (22.0%) l/min/m2, between Tpa and PCCI: -0.07 +/- 0.92 (33.5%) l/min/m2 and between Tpc and PCCI: 0.10 +/- 1.00 (35.5%) l/min/m2. For changes in CI from one time point to the next (DeltaCI), the limits of agreements between DeltaCI Tpa and DeltaCI Tpc were 0.04 +/- 0.90 l/min/m2, between DeltaCI Tpa and DeltaCI PCCI: -0.02 +/- 1.22 l/min/m2 and between DeltaCI Tpc and DeltaCI PCCI: -0.08 +/- 1.32 l/min/m2. In OPCAB surgery, limits of agreement comparing thermodilution methods were smaller than comparing PCCI with thermodilution. Recalibration of PCCI is therefore advisable.


Absence of clinical disease and contact transmission of HPAI H5NX clade 184.108.40.206 from North America in experimentally infected pigs.
In the fall of 2014, highly pathogenic avian influenza (HPAI) subtype H5N8 clade 220.127.116.11 was introduced into North America by migrating waterfowl from Asia where, through reassortment, novel HPAI H5N2 and H5N1 viruses emerged. Assess the susceptibility of pigs to HPAI H5N1, H5N2, and H5N8 clade 18.104.22.168 from North America. Pigs and trachea explants were inoculated with a representative panel of H5NX clade 220.127.116.11 HPAI viruses from North America. Nasal swabs, BALF, and sera were collected to assess replication and transmission in challenged and direct contact pigs by RRT-PCR, virus isolation, hemagglutination inhibition, and ELISA. Limited virus replication was restricted to the lower respiratory tract of challenged pigs, though absent in the nasal passages and trachea cultures, as determined by RRT-PCR in all samples. Seroconversion of inoculated pigs was detected by NP ELISA but was not reliably detected by antigen-specific hemagglutination inhibition. Boost with adjuvanted virus was required for the production of neutralizing antibodies to assess cross-reactivity between wild-type avian strains. All RRT-PCR and serology tests were negative for contact animals indicating a failure of transmission from primary inoculated pigs. H5NX clade 18.104.22.168 strains can replicate in the lower respiratory tract of swine upon high titer inoculation, though appear to be incapable of replication in swine nasal epithelium in vivo or ex vivo in trachea explants in culture. Infected pigs did not produce high levels of serum antibodies following infection. Collectively, our data show HPAI H5NX clade 18.104.22.168 viruses to be poorly adapted for replication and transmission in swine.


[Neurosurgical treatment of headaches].
Apart from headaches due to expanding lesions or vascular abnormalities which require a direct neurosurgical approach, some cases of untractable, acute or chronic headaches may need some neurosurgical management. Among these, "low pressure hydrocephalus" is responsible for a number of cases that can be relieved by ventriculo-atrial or ventriculo-peritoneal shunts: "dural headaches" including some atypic migraines and "cluster headaches" can be cured by sectioning of the afferent meningeal rootlet of the trigeminal nerve in its retrogasserian portion. Some occipital types of headache, "venous headache" or "strain headache", may benefit of a cross section of the occipital dura mater. The Authors have also used in selected cases of untractable headache, their technique of intermittent electrical stimulation of the nucleus ventralis posterolateralis by means of an implanted stimulator.


Transhumanism: How Far Is Too Far?
Transhumanism promises us freedom from the biological limitations inherent in our nature. It aims to enhance physical, emotional and cognitive capacities thus opening up new possibilities and horizons of experience. Since many transhumanist aspirations resemble those within the domain of religion, this paper compares Christian ethics to transhumanist ethics with respect to the body and the environment and offers a critique of transhumanism. Three areas of contention are discussed: the modification of our given human nature, the radical extension of our lifespans and our relationship to the natural environment. It argues that in these three areas, the underlying values being transmitted to future generations about the body and the environment are incompatible with Christian principles.


Extracellular vesicles from retinal pigment epithelial cells expressing R345W-Fibulin-3 induce epithelial-mesenchymal transition in recipient cells.
We have shown previously that expression of R345W-Fibulin-3 induces epithelial-mesenchymal transition (EMT) in retinal pigment epithelial (RPE) cells. The purpose of the current study was to determine if extracellular vesicles (EVs) derived from RPE cells expressing R345W-Fibulin-3 mutation are sufficient to induce EMT in recipient cells. ARPE-19 cells were infected with luciferase-tagged wild-type (WT)- Fibulin-3 or luciferase-tagged R345W-Fibulin-3 (R345W) using lentiviruses. EVs were isolated from the media by ultracentrifugation or density gradient ultracentrifugation. Transmission electron microscopy and cryogenic electron microscopy were performed to study the morphology of the EVs. The size distribution of EVs were determined by nanoparticle tracking analysis (NTA). EV cargo was analysed using LC-MS/MS based proteomics. EV-associated transforming growth factor beta 1 (TGFβ1) protein was measured by enzyme-linked immunosorbent assay. The capacity of EVs to stimulate RPE migration was evaluated by treating recipient cells with WT- or R345W-EVs. The role of EV-bound TGFβ was determined by pre-incubation of EVs with a pan-TGFβ blocking antibody or IgG control. EM imaging revealed spherical vesicles with two subpopulations of EVs: a group with diameters around 30 nm and a group with diameters over 100 nm, confirmed by NTA analysis. Pathway analysis revealed that members of the sonic hedgehog pathway were less abundant in R345W- EVs, while EMT drivers were enriched. Additionally, R345W-EVs had higher concentrations of TGFβ1 compared to control. Critically, treatment with R345W-EVs was sufficient to increase EMT marker expression, as well as cell migration in recipient cells. This EV-increased cell migration was significantly inhibited by pre-incubation of EVs with pan-TGFβ-neutralising antibody. In conclusion, the expression of R345W-Fibulin-3 alters the size and cargo of EVs, which are sufficient to enhance the rate of cell migration in a TGFβ dependent manner. These results suggest that EV-bound TGFβ plays a critical role in the induction of EMT in RPE cells.


Visualization of global RNA synthesis in a human (mini-) organ in situ by click chemistry.
RNA synthesis can be detected by 5-ethynyl uridine (EU) incorporation and click chemistry. Despite identifying a fundamental functional process, this technique has yet to be widely applied to complex human tissue systems. By incorporating EU into human hair follicle (HF) organs cultured ex vivo, nascent RNA synthesis was detected in situ. EU differentially incorporated across the HF epithelium. Interestingly, RNA synthesis did not correlate with protein synthesis, proliferation or epithelial progenitor cell marker expression. By treating human HFs with the cytotoxic cell cycle inhibitor (R)-CR8, which inhibits transcriptional regulators CDK7 and CDK9, it was further shown that this technique can be used to sensitively detect changes in global RNA synthesis in situ. Together, this work delineates new insights into nascent RNA synthesis within a human (mini)- organ and describes a novel read-out parameter that will enrich future ex vivo human tissue research studies.


Effects of electrostatic neutralization of Keggin Fe13 on the removal of micro and nano plastic.
The successful preparation and identification of Keggin-structure Fe13 clusters in recent years further enriched the potential application scenarios of ferric coagulants. Comparing the coagulation efficiencies and mechanisms of Fe13 in the removal of nano/microplastics with conventional polymeric Al13 and monomeric Al/Fe, this work aimed to elucidate the coagulation behaviour of Fe13 compared with the traditional mono ferric coagulant, which has the coagulation applied bottleneck of quick and violet hydrolysis. The results showed that Fe13 has a similar electrostatic neutralization potential to Al13, which could keep a positively charged species, especially in acid conditions. The Fe13 species has a selective removal potential toward the microplastics with a polar functional group like ester. Moreover, Fe13 could hydrolyze to form active sol-gel hydroxides in neutral and alkalinity conditions, which is like the behaviour of traditional monomeric Fe coagulants but seldom restabilization. The electrostatic neutralization of Fe13 could enhance the removal of nano plastic from - 25-75% compared with monomeric Fe at pH 4. The higher floc density as a monomeric Fe coagulant and better electrostatic neutralization potential of Keggin Fe13 posed a good prospect for Fe13 to replace the monomeric Fe coagulants in conventional coagulation.


Amino acid absorption and production of pancreatic hormones in non-anaesthetized pigs after duodenal infusions of a milk enzymic hydrolysate or of free amino acids.
1. Six non-anaesthetized pigs (mean body-weight 57.0 kg) were used to study the intestinal absorption of amino acids (AA) from either an enzymic hydrolysate of milk (PEP) containing a large percentage of small peptides (about 50% with less than five AA residues) and very few free AA (8%), or from a mixture of free AA with an identical pattern (AAL) infused intraduodenally in one of two amounts (55 or 110 g). Concomitant insulin and glucagon production rates were estimated. 2. Each pig was previously fitted, under anaesthesia, with an electromagnetic flow probe around the portal vein, with permanent catheters in the portal vein, the carotid artery and the duodenum. Each infusion was performed after an 18 h fasting period and each pig received each infusion. The observation period lasted for 5 h. 3. The absorption of AA was greater, more rapid and more homogeneous after PEP infusion than after AAL infusion, independent of the amount infused. 4. For the majority of AA considered individually, the absorption coefficient was higher after infusion of PEP than after that of AAL. The exceptions were methionine with a higher absorption coefficient after AAL infusion, and isoleucine, aspartic acid + asparagine and glutamic acid + glutamine with identical coefficients for both infusions. 5. Some AA, such as asparagine, ornithine, citrulline and taurine, while absent in the infusates, appeared in the portal vein in appreciable amounts after the infusion of both solutions. While a small proportion of taurine may arise from recycling of taurine-containing bile salts, it seems that the gut wall is able to synthesize all four AA. 6. Insulin production did not differ according to the nature or amount of solutions infused. Glucagon production was greater after PEP infusion.


Pupillary reactivity as an early indicator of increased intracranial pressure: The introduction of the Neurological Pupil index.
This paper introduces the 7/5/2011al Pupil index (NPi), a sensitive measure of pupil reactivity and an early indicator of increasing intracranial pressure (ICP). This may occur in patients with severe traumatic brain injury (TBI), aneurysmal subarachnoid hemorrhage, or intracerebral hemorrhage (ICH). 134 patients (mean age 46 years, range 18-87 years, 54 women and 80 men) in the intensive care units at eight different clinical sites were enrolled in the study. Pupillary examination was performed using a portable hand-held pupillometer. Patients with abnormal pupillary light reactivity had an average peak ICP of 30.5 mmHg versus 19.6 mmHg for the normal pupil reactivity population (P = 0.0014). Patients with "nonreactive pupils" had the highest peaks of ICP (mean = 33.8 mmHg, P = 0.0046). In the group of patients with abnormal pupillary reactivity, we found that the first evidence of pupil abnormality occurred, on average, 15.9 hours prior to the time of the peak of ICP. Automated pupillary assessment was used in patients with possible increased ICP. Using NPi, we were able to identify a trend of inverse relationship between decreasing pupil reactivity and increasing ICP. Quantitative measurement and classification of pupillary reactivity using NPi may be a useful tool in the early management of patients with causes of increased ICP.


[Double aortic arch in infancy and childhood].
The diagnosis of double aortic arch was suspected by oesophagography and subsequently proven by angiography in six children. The patients' age ranged from 14 days to 13 years. The diagnostic value of angiography in establishing the existence of two aortic arches is emphasized, which can easily be performed by right heart catheterization. In five patients the aorta descended to the right of the spine; the left arch was hypoplastic and became stenotic or atretic distal to the subclavian artery. In only one case the aorta descended on the left side. In this case both arches had approximately the same size. Four patients were symptomatic immediately after birth and presented with severe inspiratory stridor. The clinical course was so dramatic that surgery was inevitable at the age of four weeks, at 4, 6 and 7 months respectively. The youngest patient died postoperatively. In the other infants the stridor disappeared only months after surgery. One girl had a stridorous respiration occasionally during infancy, later on she complained of dysphagia. She was operated on at the age of 13 years. The double aortic arch was an incidental finding in a girl of 8 years; surgery was not performed.


New tumours in non-enucleated eyes of bilateral retinoblastoma patients.
Forty non-enucleated eyes with bilateral retinoblastoma which were treated by external beam radiation therapy (EBRT), transconjunctival cryopexy, and photocoagulation were retrospectively analysed for the age of onset and location of new intraocular tumours. Of these 40 eyes, 9 (22.5%) eyes developed 17 new tumour foci over a mean follow-up of 3 years. The risk of new tumour formation was age-related being 47% in children with age at onset of retinoblastoma less than 1 year compared to 4.4% in older children (P < 0.001). Four eyes (44%) had 2 episodes of tumour formation. All tumour foci developed within 11 months of initial treatment at an average episode interval of 4.0 months. In 89% of cases, new lesions ceased to occur by 18 months of age. Our study clearly shows that EBRT did not prevent development of new lesions. The tumour islands which developed in the peripheral retina in 88% of cases were successfully treated with transconjunctival cryopexy. In 8 cases (89%), the eye could be salvaged. All young bilateral retinoblastoma patients should undergo frequent periodic detailed examination of the retinal periphery with 360 degrees scleral depression to pick up new tumour lesion early and to treat them effectively with simple globe saving methods.


Decompressive craniectomy or not: intraoperative experience in 41 patients with severe traumatic brain injury.
To present our experience in using decompressive craniectomy (DC) among severe traumatic brain injury (TBI) patients during operation and to discuss its indication. From October 2008 to May 2009, 41 patients aged between 18 and 75 years with severe TBI were included in this study. They underwent DC or non-DC (NDC) according to their intraoperative findings. Postoperative intracranial pressure (ICP), complications, requiring second operation or not and outcomes were observed. Fifteen patients underwent DC and 26 patients did not. The average postoperative ICP of each patient was lower than 20 mm Hg. For patients received DC, 2 had seizures after operation and 1 developed cerebrocele in the follow-up period; only 1 NDC patient had post-traumatic seizures, but none of them had delayed haematoma, cerebrospinal fluid fistula, cerebrocele or infections. At the end of follow-up, 10 patients died, 6 had the GOS of 2, 2 of 3, 9 of 4 and 14 of 5. DC is necessary to manage fulminant intracranial hypertension or intraoperative brain swelling. If there was not brain swelling after removal of the haematoma and necrotized neural tissues, it is safe to replace skull flap. The intraoperative finding is an important factor to decide whether to perform DC or not.


Efficacy of autologous whole blood or serum therapy for chronic spontaneous urticaria: a systematic review and meta-analysis.
Background: Chronic spontaneous urticaria (CSU) is chronic wheals without identifiable exogenous stimuli. Autologous whole blood (AWB) injection and autologous serum therapy (AST) are alternative therapies for CSU that induce tolerance to circulating histamine-releasing factors. Objective: We elucidated currently available evidence for the efficacy and safety of AWB therapy and AST for CSU. Methods: We systematically searched four databases for eligible studies to perform meta-analysis. The primary outcome was the efficacy of AST or AWB therapy, and the secondary outcome was improvement after intervention based on the autologous serum skin test (ASST) status of patients. Results: Eight clinical trials, including four randomized controlled trials and 529 CSU patients, were identified. AST was not more effective than the placebo treatment in alleviating CSU symptoms at the end of treatment (p = .161), and AWB injection was also not more effective in response rates than the placebo at the end of follow-up (p = .099). Furthermore, the efficacy of AST or AWB injection for CSU and the ASST status were not significantly related. No remarkable adverse events were recorded during therapy. Conclusions: Our meta-analysis suggested that AWB therapy and AST are not significantly more effective in alleviating CSU symptoms than the placebo treatment.


Novel approaches to visualization and data mining reveals diagnostic information in the low amplitude region of serum mass spectra from ovarian cancer patients.
The ability to identify patterns of diagnostic signatures in proteomic data generated by high throughput mass spectrometry (MS) based serum analysis has recently generated much excitement and interest from the scientific community. These data sets can be very large, with high-resolution MS instrumentation producing 1-2 million data points per sample. Approaches to analyze mass spectral data using unsupervised and supervised data mining operations would greatly benefit from tools that effectively allow for data reduction without losing important diagnostic information. In the past, investigators have proposed approaches where data reduction is performed by a priori "peak picking" and alignment/warping/smoothing components using rule-based signal-to-noise measurements. Unfortunately, while this type of system has been employed for gene microarray analysis, it is unclear whether it will be effective in the analysis of mass spectral data, which unlike microarray data, is comprised of continuous measurement operations. Moreover, it is unclear where true signal begins and noise ends. Therefore, we have developed an approach to MS data analysis using new types of data visualization and mining operations in which data reduction is accomplished by culling via the intensity of the peaks themselves instead of by location. Applying this new analysis method on a large study set of high resolution mass spectra from healthy and ovarian cancer patients, shows that all of the diagnostic information is contained within the very lowest amplitude regions of the mass spectra. This region can then be selected and studied to identify the exact location and amplitude of the diagnostic biomarkers.


The cyanobacterial circadian clock follows midday in vivo and in vitro.
Circadian rhythms are biological oscillations that schedule daily changes in physiology. Outside the laboratory, circadian clocks do not generally free-run but are driven by daily cues whose timing varies with the seasons. The principles that determine how circadian clocks align to these external cycles are not well understood. Here, we report experimental platforms for driving the cyanobacterial circadian clock both in vivo and in vitro. We find that the phase of the circadian rhythm follows a simple scaling law in light-dark cycles, tracking midday across conditions with variable day length. The core biochemical oscillator comprised of the Kai proteins behaves similarly when driven by metabolic pulses in vitro, indicating that such dynamics are intrinsic to these proteins. We develop a general mathematical framework based on instantaneous transformation of the clock cycle by external cues, which successfully predicts clock behavior under many cycling environments.


Cartilage tissue from sites of weight bearing in patients with osteoarthritis exhibits a differential phenotype with distinct chondrocytes subests.
Osteoarthritis (OA) is a degenerative joint disease associated with excessive mechanical loading. The aim here was to elucidate whether different subpopulations of chondrocytes exhibit distinct phenotypes in response to variations in loading conditions. Furthermore, we seek to investigate the transcriptional switches and cell crosstalk among these chondrocytes subsets. Proteomic analysis was performed on cartilage tissues isolated from weight-bearing and non-weight-bearing regions. Additionally, single-cell RNA sequencing was employed to identify different subsets of chondrocytes. For disease-specific cells, in vitro differentiation induction was performed, and their presence was confirmed in human cartilage tissue sections using immunofluorescence. The molecular mechanisms underlying transcriptional changes in these cells were analysed through whole-transcriptome sequencing. In the weight-bearing regions of OA cartilage tissue, a subpopulation of chondrocytes called OA hypertrophic chondrocytes (OAHCs) expressing the marker genes SLC39A14 and COL10A1 are present. These cells exhibit unique characteristics of active cellular interactions mediated by the TGFβ signalling pathway and express OA phenotypes, distinct from hypertrophic chondrocytes in healthy cartilage. OAHCs are mainly distributed in the superficial region of damaged cartilage in human OA tissue, and on TGFβ stimulation, exhibit activation of transcriptional expression of iron metabolism-related genes, along with enrichment of associated pathways. This study identified and validated the existence of a subset of OAHCs in the weight-bearing area of OA cartilage tissue. Our findings provide a theoretical basis for targeting OAHCs to slow down the progression of OA and facilitate the repair of cartilage injuries.


The accuracy of HIV rapid testing in integrated bio-behavioral surveys of men who have sex with men across 5 Provinces in South Africa.
We describe the accuracy of serial rapid HIV testing among men who have sex with men (MSM) in South Africa and discuss the implications for HIV testing and prevention.This was a cross-sectional survey conducted at five stand-alone facilities from five provinces.Demographic, behavioral, and clinical data were collected. Dried blood spots were obtained for HIV-related testing. Participants were offered rapid HIV testing using 2 rapid diagnostic tests (RDTs) in series. In the laboratory, reference HIV testing was conducted using a third-generation enzyme immunoassay (EIA) and a fourth-generation EIA as confirmatory. Accuracy, sensitivity, specificity, positive predictive value, negative predictive value, false-positive, and false-negative rates were determined.Between August 2015 and July 2016, 2503 participants were enrolled. Of these, 2343 were tested by RDT on site with a further 2137 (91.2%) having definitive results on both RDT and EIA. Sensitivity, specificity, positive predictive value, negative predictive value, false-positive rates, and false-negative rates were 92.6% [95% confidence interval (95% CI) 89.6-94.8], 99.4% (95% CI 98.9-99.7), 97.4% (95% CI 95.2-98.6), 98.3% (95% CI 97.6-98.8), 0.6% (95% CI 0.3-1.1), and 7.4% (95% CI 5.2-10.4), respectively. False negatives were similar to true positives with respect to virological profiles.Overall accuracy of the RDT algorithm was high, but sensitivity was lower than expected. Post-HIV test counseling should include discussions of possible false-negative results and the need for retesting among HIV negatives.


New insights into the families of PLC enzymes: looking back and going forward.
A study in this issue of the Biochemical Journal by Harden and colleagues, in association with one published in the Biochemical Journal very recently [Hwang, Oh, Shin, Kim, Ryu and Suh (2005) Biochem. J. 389, 181-186], have defined a new member of the superfamily of PLC (phosphoinositide-specific phospholipase C) enzymes, PLCeta. Two isoforms, PLCeta1 and PLCeta2, and their splice variants add to the molecular diversity of PLC enzymes. The studies of PLCeta regulation suggest that at least some splice variants of PLCeta2 could be regulated by the G-protein subunits Gbetagamma. As two other families, PLCbeta and PLC, are also regulated through heterotrimeric G-proteins, this finding reveals further complexity and possible interplay between different PLC families and their regulatory networks. At this point, when it is likely that the PLCeta family completes the effort of identifying new members of this related group of PLC enzymes, I also discuss some more general concepts of PLC regulation and catalysis, and challenges awaiting their further studies.


[Inhibitors of proteolytic enzymes in the blood serum of patients with diphyllobothriasis].
The proteinase inhibitors alpha 1-protease inhibitor (alpha 1-antitrypsin) and alpha 2-macroglobulin have been determined in 24 patients with diphyllobothriasis (D. latum and D. dendriticum), with their activity expressed in conventional units per ml serum inhibiting or binding the activity of 1 unit of trypsin, respectively. The specific activity of alpha 1-protease inhibitor and alpha 2-macroglobulin proved to be twice (46.2%) and thrice (32.7%) lower, respectively, despite a significant decrease in the total protein serum level. The ratio between these 2 parameters was 9.86 +/- 1.4, it being equal to 5.3 +/- 0.12 in the control. The authors believe that the reduced activity of alpha 1-protease inhibitor may be a protective response of the host to the invasion. Reduced activity is suggested to promote to chronic intestinal inflammation. Decreased activity of alpha 2-macroglobulin is accounted for by damages in its synthesis by monocyte-macrophage elements and probably by enhanced binding with proteinases of the parasite.


A cocaine-regulated and amphetamine-regulated transcript inhibits oxidative stress in neurons deprived of oxygen and glucose.
Stroke, of which about 87% is ischemic stroke, constitutes one of the main causes of morbidity, disability, and mortality worldwide. Ischemic brain injury has complex pathological mechanisms. Considerable evidence has been collected over the last few years suggesting that oxidative stress associated with excessive production of reactive oxygen species is a fundamental mechanism of brain damage in stroke and reperfusion after stroke. Oxidative stress is an important trigger of neuronal apoptosis in ischemic stroke. In this current study, it was found that cocaine-regulated and amphetamine-regulated transcript 55-102 (CART55-102) inhibited oxygen-induced and glucose deprivation (OGD)-induced neurotoxicity in a dose-dependent manner. The peak dose of CART55-102 was 0.4 nmol/l. In addition, the level of intracellular reactive oxygen species was decreased in OGD-treated neurons in the presence of 0.4 nmol/l CART55-102. Mitochondrial membrane potential (ΔΨm) and mtDNA mRNA expressions were increased in OGD-treated neurons in the presence of 0.4 nmol/l CART55-102. The current study suggests that CART55-102, by inhibiting oxidative stress, may be developed into therapeutic agents for ischemic stroke.


Parameterization of the three-dimensional room transfer function in horizontal plane.
This letter proposes an efficient parameterization of the three-dimensional room transfer function (RTF) which is robust for the position variations of source and receiver in respective horizontal planes. Based on azimuth harmonic analysis, the proposed method exploits the underlying properties of the associated Legendre functions to remove a portion of the spherical harmonic coefficients of RTF which have no contribution in the horizontal plane. This reduction leads to a flexible measuring-point structure consisting of practical concentric circular arrays to extract horizontal plane RTF coefficients. The accuracy of the above parameterization is verified through numerical simulations.