Interleukin 3, granulocyte-macrophage colony-stimulating factor, and transfected erythropoietin receptors mediate tyrosine phosphorylation of a common cytosolic protein (pp100) in FDC-ER cells.
Receptors for the hematopoietic growth factors erythropoietin, interleukin 3 (IL-3), and granulocyte-macrophage colony-stimulating factor (GM-CSF) are members of a structurally related receptor superfamily. Interestingly, while none of these receptors encode tyrosine kinase activities, induced tyrosine phosphorylation has been observed in various responsive cells stimulated with each factor. Toward defining possible common transduction pathways which are activated by these three cytokines, we have studied induced protein phosphorylation in murine myeloid FDC-P1 cells stably transfected with an erythropoietin receptor cDNA (FDC-ER cells). FDC-ER cells proliferate in response to erythropoietin (Quelle, D. E., and Wojchowski, D. M. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 4801-4805), and presently are shown to rapidly phosphorylate a M(r) 100,000 cytosolic protein (pp100) at tyrosine residues in response to this factor. Phosphorylation of pp100 also is induced in FDC-P1 and FDC-ER cells in response to IL-3 or GM-CSF. Importantly, quantitative analyses showed identical concentration dependencies for factor-induced pp100 phosphorylation and induced cell proliferation. Moreover, a selective loss of proliferative responsiveness to GM-CSF in FDC-ER cells was associated with a reduced capacity of GM-CSF to induce pp100 phosphorylation. Finally, limited differences in tryptic phosphopeptide maps of pp100 as isolated following exposure to erythropoietin, IL-3, or GM-CSF were observed, suggesting that these factors also may preferentially induce phosphorylation of pp100 at distinct sites. These findings are consistent with a role for pp100 as a common cytosolic transducer in the apparently convergent pathways of erythropoietin-, IL-3-, and GM-CSF-induced proliferation of myeloid progenitor cells.


Identification by monoclonal antibodies of a new epitope in the glycoprotein complex of Sindbis virus.
Monoclonal antibodies against a deletion mutant of Sindbis virus were produced and characterized in order to determine the fine mapping and functional activities of single viral epitopes. All monoclonal antibodies so far tested showed a certain degree of reciprocal competition and were directed against an antigenic determinant which was present only on the undissociated complex of the E1 and E2 glycoproteins. A biological assay measuring viral haemagglutination showed no decrease in the titre of viral samples preincubated with monoclonal antibodies. Conversely, a reduction in viral infectivity was demonstrated, particularly with two of these antibodies. The results suggest that the antibodies which we characterized seem to recognize a new epitope which is represented on both glycoproteins on the surface of this mutant of Sindbis virus.


Methodology guideline for clinical studies investigating traditional Chinese medicine and integrative medicine: executive summary.
This guideline aims to provide a methodological guidance for clinical studies in TCM and integrative medicine in terms of study design, execution, and reporting. The commonly used methods including experimental and observational methods were introduced in this guideline such as randomized clinical trials, cohort study, case-control study, case series, and qualitative method which can be incorporated into above quantitative methods. The guideline can be used for the evaluation of therapeutic effect of TCM therapies or their combination with conventional therapy. TCM therapy refers to one of the followings or their combination: herbal medicine, acupuncture, moxibustion, cupping, Taichi/Qigong, and Guasha,Tuina (therapeutic massage). It is also suitable for research and development of ethnopharmaceuticals or folk medicine.


Hot or not? Discovery and characterization of a thermostable alditol oxidase from Acidothermus cellulolyticus 11B.
We describe the discovery, isolation and characterization of a highly thermostable alditol oxidase from Acidothermus cellulolyticus 11B. This protein was identified by searching the genomes of known thermophiles for enzymes homologous to Streptomyces coelicolor A3(2) alditol oxidase (AldO). A gene (sharing 48% protein sequence identity to AldO) was identified, cloned and expressed in Escherichia coli. Following 6xHis tag purification, characterization revealed the protein to be a covalent flavoprotein of 47 kDa with a remarkably similar reactivity and substrate specificity to that of AldO. A steady-state kinetic analysis with a number of different polyol substrates revealed lower catalytic rates but slightly altered substrate specificity when compared to AldO. Thermostability measurements revealed that the novel AldO is a highly thermostable enzyme with an unfolding temperature of 84 °C and an activity half-life at 75 °C of 112 min, prompting the name HotAldO. Inspired by earlier studies, we attempted a straightforward, exploratory approach to improve the thermostability of AldO by replacing residues with high B-factors with corresponding residues from HotAldO. None of these mutations resulted in a more thermostable oxidase; a fact that was corroborated by in silico analysis.


Mechanism of action of anti-HER2 monoclonal antibodies: scientific update on trastuzumab and 2C4.
The HER family of transmembrane tyrosine kinase receptors is composed of four members, BER1 to HER4. HER2 is a ligand-orphan receptor expressed in many human tumors and overexpressed in 25-30% of breast cancers. HER2 amplifies the signal provided by other receptors of the HER family by forming heterodimers. The essential role of HER2 in the HER signaling network led to the development of anti-HER2 monoclonal antibodies (MAbs) for cancer therapy. In particular, the humanized MAb trastuzumab (Herceptin) has antitumor activity against HER2-overexpressing human breast tumor cells and is widely used for the treatment of women with HER2 overexpressing breast cancers. Trastuzumab induces HER2 receptor downmodulation and, as a result, inhibits critical signalling pathways (i.e. ras-Raf-MAPK and PI3K/Akt) and blocks cell cycle progression by inducing the formation of p27/Cdk2 complexes. Trastuzumab also inhibits HER2 cleavage, preceding antibody-induced receptor downmodulation, and this effect might contribute to its antitumor activity in some cancers. In vivo, trastuzumab inhibits angiogenesis and induces antibody-dependent cellular cytotoxicity. A limitation of trastuzumab is that its activity is largely restricted to breast cancers with the highest level of HER2 overexpression or HER2 gene amplification. However, there is a large population of breast cancers and of many other tumors that have low or moderate HER2 expression. In such tumors, HER2 functions as a preferred coreceptor to form heterodimers with HER1 (EGFR), HER3 or HER4. For this reason, a humanized monoclonal antibody, called 2C4, that targets the role of HER2 as a coreceptor is under active development. 2C4 binds to a different epitope of HER2 ectodomain than trastuzumab and sterically hinders HER2 recruitment in heterodimers with other HER receptors. This results in the inhibition of signalling by HER2-based heterodimers both in cells with low and high HER2 expression. In vitro and in vivo antitumor activity has been reported in a range of breast and prostate tumor models. Therefore, 2C4 may have potential against a wide variety of solid tumors. Phase I trials are underway.


How organizational context affects bioethical decision-making: pharmacists' management of gatekeeping processes in retail and hospital settings.
Social science studies of bioethics demonstrate that ethics are highly contextual, functioning differently across local settings as actors make daily decisions "on the ground." Sociological studies that demonstrate the key role organizations play in shaping ethical decision-making have disproportionately focused on physicians and nurses working in hospital settings where they contend with life and death issues. This study broadens our understanding of the contexts of ethical decision-making by empirically examining understudied healthcare professionals - pharmacists - working in two organizational settings, retail and hospital, where they act as gatekeepers to regulated goods and services as they contend with ethical issues ranging from the serious to the mundane. This study asks: How do organizations shape pharmacists' identification, negotiation, and resolution of ethical challenges; in other words, how do organizations shape pharmacists' gatekeeping processes? Based on 95 semi-structured interviews with U.S. pharmacists practicing in retail and hospital pharmacies conducted between September 2009 and May 2011, this research finds that organizations influence ethical decision-making by shaping how pharmacists construct four gatekeeping processes: medical, legal, fiscal, and moral. Each gatekeeping process manifests differently across organizations due to how these settings structure inter-professional power dynamics, proximity to patients, and means of accessing information. Findings suggest new directions for theorizing about ethical decision-making in medical contexts by drawing attention to new ethical actors, new organizational settings, an expanded definition of ethical challenges, and a broader conceptualization of gatekeeping.


A robust characterization of retinoic acid response elements based on a comparison of sites in three species.
The availability of high-throughput genomic sequencing has allowed us to construct a more robust characterization of retinoic acid response elements than was possible in the past. We located human, mouse, and rat homologs for each of 51 well-documented, conserved retinoic acid response elements. Mathematical and statistical analyses of these 153 sites, 78 of which are new, shows that 92% of response elements have direct-repeat symmetry, but that only 76% exhibit canonical spacing attributes. While the familiar '(a/g)g(g/t)tca' hexamer motif is upheld, the more relaxed sequence, '(a/g)g(g/t)(g/t)(g/c)a', represents a 10% consensus. Sites are as likely to be on the coding strand as on the non-coding strand, and 86% of them are in upstream locations. From a statistical point of view, DR1 elements are fundamentally different from DR2 and DR5 elements, but this is only evident in the 5' hexamer. While there is considerable variation in core positions, and while no nucleotide can be considered forbidden at any position, variation among species at a fixed locus appears surprisingly constrained once a functional site has been attained.


A sequence-dependent 1H-NMR study on the formation of beta-turns in tetrapeptides containing charged residues.
The importance of side-chain charge interactions in the formation of beta-turns was studied. Sixteen protected NAc-tetrapeptide amides were studied, namely the variants of DEKS: NEKS, EEKS, DDKS, DQKS, NQKS, DERS, NERS, EERS, DDRS, NDRS, DQRS, and DKES. Three tetrapeptides--NPDM, NSDM, and NDDS--were also studied as they have a high probability of forming beta-turns, based on statistical predictions. The results indicate that a small proportion of type I beta-turn exists in solutions of DEKS and DERS in methanol/water (60/40), while NEKS has an even smaller population of this turn. The other tetrapeptides are present in solution only in the extended conformation. These results clearly show the importance of the salt bridge between the side chains of K2 and E3 or R2 and E3, as well as the importance of the charge on the side chain of the first residue in stabilizing the beta-turn. The relevance of statistical predictions for beta-turns in short peptides is discussed.


Background clutter amplitude and frequency of GPR signals to analyse water content in sedimentary deposits: Urban infrastructure environment.
Soil site studies are crucial in the analysis of seismic hazard in populated areas. This study focusses on the use of the Ground Penetrating Radar as a non-destructive geophysical method to analysis the water content of a sedimentary basin of a local urban area in Cervantes Parks in Barcelona city. Main objective of this work is to understand further the local seismic soil site, and analysis the background clutter amplitude and frequency of the GPR signal applied on the seasonal changes of the underground water content. Changes produces in the GPR signal could be associated to the changes of the ground material due to the existence of water content and water flow, which influence the grain size distribution of the soil characterisation. Previously, several methodologies have been applied to determine the changes of the ground material due to the existence of water content and most importantly effect of the seasonal changes on the soil characterisation. GPR has been applied as a new technology and non-invasive to further analysis water content in shallow geology. Methodology applied on this study compromised in four different phases; Two seasonal data collection using 100 MHz centre frequency shielded antenna in both dry and wet season, radar images are proceeded with common post-processing approaches, following more innovative processing approach based on the Fast Fourier Transform (FFT) method in order to analysis background clutter amplitude and frequency spectrum to achieve an accurate interpretation of underground water content activities, in conclusion a comparative study of the results driven from radar images are elaborated considering historical knowledge. Results show that GPR is an efficient technique to analysis water content invasively considering low frequency antennas to achieve deeper penetration depth in sedimentary deposits, and predict shifting and increasing of the underground streams due to changes in weather conditions. Results could contribute to the enhancing local seismic site behaviour in populated areas considering continuous monitoring of underground water activities in sedimentary basin sites.


Efficacy and safety of long-acting growth hormone in adult growth hormone deficiency: A systematic review and meta-analysis.
No meta-analysis has analysed efficacy and safety of long-acting growth hormone (GH) therapy in adult GH deficiency. We undertook this meta-analysis to address this gap in knowledge METHODS: Electronic databases were searched for RCTs involving adult GH deficiency patients receiving weekly long-acting GH as compared to daily GH/placebo controls. Primary outcome was to evaluate changes in body-composition parameters. Secondary outcomes were to evaluate alterations in glycaemia and adverse-events. Data from 5 studies involving 648 patients were analysed (4 studies having daily GH as active controls; 1 study having placebo as passive controls). Over 24-34 weeks clinical use, patients receiving long-acting GH had comparable change in lean mass [MD-0.28 kg (95%CI: 0.94 - 0.38); P = 0.41; I2 = 29% (low heterogeneity)] and fat mass [MD-0.10 kg (95%CI: 1.97-1.78); P = 0.92; I2 = 77%(considerable heterogeneity)] as compared to daily GH injections. Long-acting GH use was associated with significantly lower visceral adipose tissue [MD-1.75 cm2(95%CI: 2.14 to -1.35); P < 0.01; I2 = 0% (low heterogeneity)] and higher gynoid fat-mass [MD 0.14 kg(95%CI:0.02-0.26); P = 0.03] compared to daily GH injections. Total adverse events [Risk ratio (RR) 1.65 (95% CI: 0.83-3.29); P = 0.15; I2 = 68%] and severe adverse events [RR 0.60 (95% CI: 0.30-1.19); P = 0.14; I2 = 0%] were not significantly different in long-acting GH group compared to controls. Occurrence of headache, arthralgia, nasopharyngitis, new onset diabetes, anti-GH antibodies were comparable among groups. Long-acting GH users had significantly higher treatment adherence compared to controls [OR 4.80 (95%CI:3.58-6.02); P < 0.01; I2 = 0%]. Long-acting GH has comparable beneficial impact on body composition parameters in adult GH deficiency, is well tolerated without any increased adverse events.


Protein profiling of rat ventral prostate following chronic finasteride administration: identification and localization of a novel putative androgen-regulated protein.
To better understand the effects of antiandrogens on the prostate, we investigated the changes in the proteome of rat ventral prostate (VP) following treatment with a well characterized 5alpha-reductase inhibitor, finasteride. Sprague-Dawley rats were treated daily by gavage with finasteride at 0, 1, 5, 25, and 125 mg/kg/day. Changes in plasma hormone levels as well as the weight and histology of sex accessory tissues were determined after 28 days of treatment and showed a dose-related decrease of VP weights together with a marked atrophy of the tissue visible at the macroscopic and microscopic levels. In addition, significant reductions in seminal vesicle and epididymis weights were noted. VP proteins were analyzed by two-dimensional gel electrophoresis: 37 proteins, mainly involved in protein synthesis, processing, and cellular trafficking and in metabolism, detoxification, and oxidative stress, were identified as modulated by finasteride. The prominent feature of this study is the demonstration of finasteride dose-dependent up-regulation of a protein similar to l-amino-acid oxidase 1 (Lao1). An up-regulation of this protein was also observed with the antiandrogen flutamide. Lao1 expression occurred as early as 48 h after antiandrogen administration and persisted throughout the treatment duration. Immunohistochemistry showed that this protein was only detectable in epithelial cells and secretory vesicles. Altogether these data point to a potential use of Lao1 to reveal antiandrogen-induced prostate injury.


The impact of comorbidities on the physical function in patients with rheumatoid arthritis.
To investigate the association of comorbidities with mobility limitation and functional disability in patients with rheumatoid arthritis and to identify which comorbidity indicator is the most appropriate to determine this association. Sixty rheumatoid arthritis patients were enrolled in a cross-sectional study for a period of 11 months. Comorbidities were assessed using three indicators: (i) the total number of comorbidities; (ii) the Charlson comorbidity index; and (iii) the functional comorbidity index. Disease activity was assessed using the Disease Activity Score 28. Functional capacity was measured using the Health Assessment Questionnaire, and mobility was measured using Timed Up and Go Test and Five-Times-Sit-to-Stand Test. Statistical analysis was performed using a stepwise log-linear multiple regression with a significance level of 5%. In the final model, only comorbidity was associated with mobility limitation. The functional comorbidity index score explained 19.1% of the variability of the Five-Times-Sit-to-Stand Test (coefficient of determination [R(2)]=0.191) and 19.5% of the Timed Up and Go Test variability (R(2)=0.195). With regard to functional disability, the associated factors were comorbidity and disease activity, which together explained 32.9% of the variability of the Health Assessment Questionnaire score (adjusted R(2)=0.329). Comorbidities were associated with mobility limitation and functional disability in rheumatoid arthritis patients. The functional comorbidity index proved to be an appropriate comorbidity indicator to determine this association.


Gender-specific differences in clinical outcome of primary prevention implantable cardioverter defibrillator recipients.
To assess differences in clinical outcome of implantable cardioverter-defibrillator (ICD) treatment in men and women. Prospective cohort study. University Medical Center. 1946 primary prevention ICD recipients (1528 (79%) men and 418 (21%) women). Patients with congenital heart disease were excluded for this analysis. All-cause mortality, ICD therapy (antitachycardia pacing and shock) and ICD shock. During a median follow-up of 3.3 years (25th-75th percentile 1.4-5.4), 387 (25%) men and 76 (18%) women died. The estimated 5-year cumulative incidence for all-cause mortality was 20% (95% CI 18% to 23%) for men and 14% (95% CI 9% to 19%) for women (log rank p<0.01). After adjustment for potential confounding covariates all-cause mortality was lower in women (HR 0.65; 95% CI 0.49 to 0.84; p<0.01). The 5-year cumulative incidence for appropriate therapy in men was 24% (95% CI 21% to 28%) as compared with 20% (95% CI 14% to 26%) in women (log rank p=0.07). After adjustment, a non-significant trend remained (HR 0.82; 95% CI 0.64 to 1.06; p=0.13). In clinical practice, 21% of primary prevention ICD recipients are women. Women have lower mortality and tend to experience less appropriate ICD therapy as compared with their male peers.


Adult Wilms tumor treated with radiotherapy and chemotherapy: a case report.
A 24-year-old man underwent a left radical nephrectomy for a mass that was found to be a Wilms tumor. The postoperative course, consisting of combined radiotherapy and chemotherapy, is discussed and the recent literature is reviewed.


Purification and characterization of detergent-compatible protease from Aspergillus terreus gr.
The possibility of using Aspergillus terreus protease in detergent formulations was investigated. Sodium dodecyl sulfate (SDS) and native polyacrylamide gel electrophoresis indicated that the purified alkaline protease (148.9 U/mg) is a monomeric enzyme with a molecular mass of 16 ± 1 kDa. This was confirmed by liquid chromatography-mass spectrometry. The active enzyme degraded the co-polymerized gelatin. The protease demonstrated excellent stability at pH range 8.0-12.0 with optimum at pH 11.0. It was almost 100 % stable at 50 °C for 24 h, enhanced by Ca2+ and Mg2+, but inhibited by Hg2+, and strongly inhibited by phenylmethyl sulfonyl fluoride. It showed maximum activity against casein followed by gelatin; its Vmax was 12.8 U/ml with its corresponding KM of 5.4 mg/ml. The proteolytic activity was activated by Tween-80, Triton-100 and SDS, and remained unaltered in the presence of H2O2 and NaClO. The enzyme exhibited higher storage stability at 4, 28 and -20 °C. It was stable and compatible to the desired level in the local detergents. The addition of the protease to the Super wheel improved its blood stain removal. The isolated protease can thus be a choice option in detergent industry.


Myocardial pro-inflammatory cytokine expression and cellular rejection in pediatric heart transplant recipients.
Accumulating evidence suggests that immune-mediated injury is important in the development of rejection after heart transplantation. We hypothesized that pro-inflammatory cytokine expression would increase in biopsy samples that manifest cellular rejection and that this would correlate with the development and progression of transplant cellular rejection. Children with heart transplants were prospectively enrolled from July 2004 to November 2005. Right ventricular endomyocardial biopsies were obtained during routine catheterization for rejection surveillance. Cellular rejection was graded using criteria established by the International Society for Heart and Lung Transplantation. RNA was extracted from biopsy samples and reverse transcription was used for complementary DNA synthesis. The cDNA product was evaluated by quantitative real-time polymerase chain reaction (PCR) to measure the following cytokines: interleukin (IL)-1beta, IL-6 and IL-18; tumor necrosis factor-alpha (TNF-alpha); and interferon-gamma (IFN-gamma). Normalized cytokine mRNA transcripts were correlated with cellular rejection scores and the presence of viral genome. Seventy-four children (mean age 9.6 +/- 5.5 years, range 0.2 to 20.5 years) were enrolled and 95 biopsies were obtained. None of the cytokines demonstrated a correlation with the cellular rejection score, even within individual patients for whom multiple, serial biopsy samples were studied. Eighteen biopsy samples were found to have parvovirus B19 genome present, but there was no correlation between cytokine levels and the presence of parvovirus. Cytokine transcripts in heart transplants do not correlate with cellular rejection. In addition, there is no correlation between cytokine transcripts and the presence of viral genome.


Flavobacterium columnare genomovar influences mortality in channel catfish (Ictalurus punctatus).
Flavobacterium columnare, causal agent of columnaris disease, is pathogenic to many species of freshwater fish throughout the world. The United States channel catfish (Ictalurus punctatus) aquaculture industry is severely impacted by columnaris disease. The majority of the F. columnare isolates recovered from diseased channel catfish belonged to either genomovars I or II. The objective of the present study was to determine if differences existed in the ability of these genomovars to induce mortality in channel catfish. Single strand conformation polymorphism analysis (SSCP) was used to ascribe the isolates used in this study to the appropriate genomovar. Immersion challenge experiments (15min immersion exposure to approximately 5x10(5) to 1x10(6) CFU/mL) were carried out to assess virulence of genomovar I and II isolates to channel catfish. The results demonstrated that genomovar II (n=4) isolates were significantly (P<0.05) more virulent to channel catfish fry (92-100% mortality) than genomovar I (n=3) isolates (0-46% mortality). In vivo adhesion of the genetically characterized F. columnare also correlated (r2=0.73) to increased mortality in the challenged fry. In fingerling channel catfish, significantly higher mortality (P<0.05) resulted with genomovar II isolates ALM-05-182 and ALG-00-530 as compared to all the genomovar I isolates (n=3). Mortality of genomovar II isolate BGFS-27 with similar to genomovar II isolate (ALG-00-530) and two genomovar I isolates (ALM-05-53 and 140). The results suggest that although both genomovars are present in the aquatic environment, genomovar II appears to be more pathogenic for channel catfish.


Immunocytochemical study of calretinin and calbindin D-28K expression in the retina of three cartilaginous fishes and a cladistian (Polypterus).
The distribution of two calcium-binding proteins, calbindin D-28K (CB) and calretinin (CR) was studied in the retina of a cladistian, Polypterus senegalus, and three cartilaginous fishes (Scyliorhinus canicula, Raja undulata and Torpedo marmorata). Western blot analysis of brain extracts revealed the lack of cross-reactivity of the used antibodies. In Polypterus, CB and CR immunoreactivities were observed in some amacrine and ganglion cells, but scarce cells showed CR/CB colocalization. Furthermore, CR immunoreactivity was present in a number of displaced bipolar cells and in some putative displaced ganglion cells, whereas CB immunoreactivity was found in some cones. No positive retinal structure was observed with the CB antibody used in cartilaginous fishes. Instead, CR was expressed in some amacrine, horizontal and ganglion cells of the dogfish and skate and, in some ganglion cells of the electric ray. The comparative analysis suggests, (1) the presence of CB-positive photoreceptor cells in the retina of cladistians seems to be apomorphic (in jawed fishes) in contrast with the plesiomorphic condition of this character in land vertebrates; (2) the presence of CR in amacrine and ganglion cells is a conserved feature along vertebrate phylogeny, whereas its variable expression in bipolar and horizontal cells represents a derived character; (3) the absence of CB in horizontal cells in cladistians could represent a derived character; and (4) the presence of CR displaced bipolar and putative displaced ganglion cells in Polypterus is shared with basal groups of actinopterygians.


Crystallographic symmetry and coenzyme binding properties of D-glyceraldehyde-3-phosphate dehydrogenase from the tail muscle of Palinurus vulgaris.
Crystals of apo- and holo-D-glyceraldehyde-3-phosphate dehydrogenase from the tail muscle of the Mediterranean lobster Palinurus vulgaris, previously found to be suitable for single crystal microspectrophotometric studies of catalytic activity in the crystalline state, have been examined by x-ray crystallography. The two forms are isomorphous, space group C 2 with cell dimensions a=128.4 A, b=99.9 A, c=80.3 A,beta=113.4 degrees. These data are consistent with a molecular weight of 73,000 in the crystallographic asymmetric unit, indicating that the tetrameric molecule possesses an exact 2-fold axis both in the presence and in the absence of NAD+. Analysis of the intensity distribution of conventional x-ray precession photographs shows that two further noncrystallographic diads are present and that the molecule has the 2 pseudo 22 symmetry found in other D-glyceraldehyde-3-phosphate dehydrogenases. Binding of NAD+ to apoenzyme in solution, at 25 degrees C, is anticooperative and it can be satisfactorily described by assuming two classes of coenzyme binding sites.


Stability of astaxanthin-loaded nanostructured lipid carriers as affected by pH, ionic strength, heat treatment, simulated gastric juice and freeze-thawing.
Nanostructured lipid carriers (NLCs) offer many potential benefits for incorporating lipophilic molecules into clear/opaque food systems. In this study, the influence of pH, ionic strength, thermal treatment, simulated gastric juice (SGJ), and freeze-thawing on the stability of astaxanthin-loaded NLCs (Ax-NLCs) was examined. Ax-NLCs (containing α-tocopherol and ethylenediaminetetraacetic acid as antioxidants) were stabilized with Tween 80 and lecithin (Z-average size: 94 nm), and studied for the above mentioned purpose. The size of Ax-NLCs increased at low pH values (≤5), high NaCl concentrations (≥50 mM), and slightly at SGJ, mainly because of decreasing the ζ-potential. Moreover, thermal treatment at 80/90 °C led to an increase in Ax-NLCs size. Glycerol was found as an appropriate cryoprotectant for preventing aggregation of Ax-NLCs during freeze-thawing. pH, ionic strength, heat and SGJ had no drastic effects on the chemical stability of astaxanthin in NLCs. These results have valuable implications for the utilization of food-grade NLCs.


On the Road to Regeneration: "Tools" and "Routes" Towards Efficient Cardiac Cell Therapy for Ischemic Cardiomyopathy.
Cardiac regenerative medicine is a field bridging together biotechnology and surgical science. In this review, we present the explored surgical roads to cell delivery and the known effects of each delivery method on cell therapy efficiency. We also list the more recent clinical trials, exploring the safety and efficacy of delivery routes used for cardiac cell therapy approaches. There is no consensus in defining which way is the most suitable for the delivery of the different therapeutic cell types to the damaged heart tissue. In addition, it emerged that the "delivery issue" has not been systematically addressed in each clinical trial and for each and every cell type capable of cardiac repair. Cardiac damage occurring after an ischemic insult triggers a cascade of cellular events, eventually leading to heart failure through fibrosis and maladaptive remodelling. None of the pharmacological or medical interventions approved so far can rescue or reverse this phenomenon, and cardiovascular diseases are still the leading cause of death in the western world. Therefore, for nearly 20 years, regenerative medicine approaches have focused on cell therapy as a promising road to pursue, with numerous preclinical and clinical testing of cell-based therapies being studied and developed. Nonetheless, consistent clinical results are still missing to reach consensus on the most effective strategy for ischemic cardiomyopathy, based on patient selection, diagnosis and stage of the disease, therapeutic cell type, and delivery route.


The potential of nanofibers to increase solubility and dissolution rate of the poorly soluble and chemically unstable drug lovastatin.
Polymer nanofibers represent a promising drug delivery system. However, as a large surface area is exposed to external conditions during the electrospinning process, they are usually used to incorporate drugs with good oxidative stability. Here, we introduce a new concept for the incorporation of a drug with low oxidative stability and extremely low solubility into poloxamer 188/poly(ethylene oxide) and Soluplus/poly(ethylene oxide) nanofibers, to improve its solubility and increase its dissolution rate. The electrospun products showed different morphologies and lower initial lovastatin contents than theoretically expected, which indicated oxidative drug degradation during electrospinning. The addition of antioxidants improved the lovastatin chemical stability in the nanofibers. The highest lovastatin dissolution rate and solubility were obtained for the Soluplus-based nanofibers, where no crystalline lovastatin was detected. The accelerated stability study has revealed the chemical stability of lovastatin in the poloxamer-based nanofibers with the addition of antioxidants, whereas in the Soluplus-based nanofibers lovastatin was not completely preserved. However, appropriate packaging of the formulation and storage under normal conditions is expected to assure its stability. These Soluplus-based nanofibers developed here with antioxidants represent a promising immediate release formulation to provide improved solubility and dissolution rate for poorly soluble and chemically unstable drugs, such as lovastatin.


Genotoxicity of polar fractions from a herbicide-contaminated soil does not correspond to parent contaminants.
Fractionation with biological and chemical analysis has been proposed to characterize the risk associated with complex samples. Thus, we have employed hot pressurized water for fractionating a herbicide-contaminated soil. Genotoxicity was found in polar and midpolarity fractions extracted from a historically herbicide-contaminated soil. However, the parent herbicides as well as organic solvent extracts (consisting of herbicides) were not genotoxic. Thus, polar organics formed by natural degradation of herbicides may be responsible for the genotoxic response. For correct assessment of the toxicity of complex matrices such as soils, both contaminated and uncontaminated samples need to be studied for their toxicological impact.


Androgen receptor YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration.
X-linked spinal and bulbar muscular atrophy (SBMA) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration. SBMA is caused by polyglutamine repeat expansions in the androgen receptor (AR). To determine the basis of AR polyglutamine neurotoxicity, we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells. The AR100 transgenic mice developed a late-onset, gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration, indicating striking recapitulation of the human disease. We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein (CBP)-mediated transcription of vascular endothelial growth factor (VEGF) and observed altered CBP-AR binding and VEGF reduction in AR100 mice. We found that mutant AR-induced death of motor neuron-like cells could be rescued by VEGF. Our results suggest that SBMA motor neuronopathy involves altered expression of VEGF, consistent with a role for VEGF as a neurotrophic/survival factor in motor neuron disease.


Self-crosslinked fibrous collagen/chitosan blends: Processing, properties evaluation and monitoring of degradation by bi-fluorescence imaging.
Porous collagen/chitosan scaffolds with different Collagen:Chitosan (Coll:Ch) ratios were prepared by freeze-drying followed by self-crosslinking via dehydrothermal treatment (DHT) and characterized as biomaterials for tissue engineering. Cy7 and Cy5.5 fluorochromes were covalently grafted to collagen and chitosan, respectively. Thus, it was possible, using optical fluorescence imaging of the two fluorochromes, to simultaneously track their in vivo biodegradation, in a blend scaffold form. The fluorescence signal evolution, due to the bioresorption, corroborated with histological analysis. In vitro cytocompatibility of Coll:Ch blend scaffolds were evaluated with standardized tests. In addition, the scaffolds showed a highly interconnected porous structure. Extent of crosslinking was analyzed by convergent analysis using thermogravimetry, Fourier Transform Infrared Spectroscopy and PBS uptake. The variations observed with these techniques indicate strong interactions between collagen and chitosan (covalent and hydrogen bonds) promoted by the DHT. The mechanical properties were characterized to elucidate the impact of the different processing steps in the sample preparation (DHT, neutralization and sterilization by β-irradiation) and showed a robust processing scheme with low impact of Coll:Ch composition ratio.


Environmental samples of microplastics induce significant toxic effects in fish larvae.
Microplastics (MPs) are present throughout aquatic ecosystems, and can be ingested by a wide variety of organisms. At present, the physical and chemical effects of environmental MPs on aquatic organisms are poorly documented. This study aims to examine the physiological and behavioral effects caused by fish consuming environmental microplastics at different life stages. MP samples were collected from beaches on three islands (Easter Island, Guam and Hawaii) located near the North and South gyres of the Pacific Ocean. Larvae and juveniles of Japanese Medaka were fed for 30days with three doses of MPs (0.01, 0.1 and 1% w/w in fish food) approximate to the concentrations measured in moderately and heavily contaminated ocean areas. Ingestion of MPs by medaka larvae caused (variously) death, decreased head/body ratios, increased EROD activity and DNA breaks and, alterations to swimming behavior. A diet of 0.1% MPs was the most toxic. Two-month-old juveniles fed with 0.01% MPs did not exhibit any symptoms except an increase in DNA breaks. Our results demonstrate ingestion and mainly sublethal effects of environmental MPs in early life stages of fish at realistic MP concentrations. The toxicity of microplastics varies from one sample to another, depending on polymer composition, weathering and pollutant content. This study examines the ecological consequences microplastic build-up in aquatic ecosystems, more particularly in coastal marine areas, which serve as breeding and growing grounds for a number of aquatic species.


Does acupuncture have advantages in the rehabilitation of vascular mild cognitive impairment? A systematic review and meta-analysis.
Vascular mild cognitive impairment (VMCI) is a common impairment caused by vascular factors. VMCI often occurs after stroke, and it is the main clinical manifestation of long-term disability. Many patients are treated with acupuncture in combination with other therapies. However, evidence regarding the effectiveness of this treatment regimen is lacking. This meta-analysis aimed to evaluate the efficacy of acupuncture therapy for treating VMCI. This systematic review was conducted in accordance with the preferred reporting and meta-analysis guidelines. The CNKI, Wanfang, VIP, CBM, Cochrane Library, PubMed and Embase databases were searched from inception to August 20, 2022. After two researchers independently screened the literature, they extracted the data and evaluated the risk of bias in the included studies. Revman 5.3 software was used for the meta-analysis. Thirty-two randomized controlled trials (RCTs) were included. The overall effective rate of acupuncture for treating VMCI was 3.06, 95% CI [2.39, 3.91], (P < 0.05). Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE), Barthel Index and Activities of Daily Living (ADLs) scores significantly differed between the treatment and control groups, with weighted mean differences (WMDs) [95% CI] (P value) of 1.97 [1.44, 2.49] (P < 0.05), 2.02 [1.50, 2.54] (P < 0.05), 5.54 [3.81, 7.28] (P < 0.05), and 3.43 [2.53, 4.33] (P < 0.05), respectively. The overall effective rate of electroacupuncture (EA) for treating VMCI was better than that of the control group (RR = 2.25, 95% CI, [1.13, 4.50], P < 0.05). MoCA, MMSE, Barthel index and ADL scores differed significantly between the treatment and control groups, with WMDs [95% CI] (P value) of 1.79 [1.20, 2.38] (P < 0.05), 1.45 [0.87, 2.03] (P < 0.05), 5.78 [2.38, 9.18] (P < 0.05), and 3.15 [2.15, 4.15] (P < 0.05), respectively. Acupuncture alone and combined with drug therapy were thus superior to drug therapy alone for improving cognitive function. EA also has potential advantages. Acupuncture combined with another therapy is better than other therapies alone, such as simple drug therapy, for treating VMCI. However, variations in study duration (4-12 weeks) limit us from drawing any definitive conclusions about long-term effects. Therefore, more RCTs with rigorous designs and reasonable treatment and follow-up durations are needed.


Equine Platelet-Rich Plasma.
Platelet-rich plasma (PRP) is an orthobiologic therapy composed of platelets, leukocytes, red blood cells, and plasma proteins. PRP has been used for 20 years, but progress determining efficacy has been slow. The definitions and classification of PRP are reviewed, and the use of PRP for tendon, ligament, and joint disease is discussed with a focus on findings of basic science and clinical studies, platelet activation, concurrent administration of nonsteroidal anti-inflammatory drugs, and treatment complications. Finally, the advantages of platelet lysates and freeze-dried platelets are discussed. The promising results of a PRP lysate optimized for antibiofilm and antimicrobial properties are introduced.


Carvacrol protects mice against LPS-induced sepsis and attenuates inflammatory response in macrophages by modulating the ERK1/2 pathway.
Macrophages play an important role in the development of life-threatening sepsis, which is characterized by multiorgan dysfunction, through their ability to produce inflammatory cytokines. Carvacrol is a phenolic compound that has been confirmed to possess strong anti‑inflammatory activity. In this study, we mainly investigated the effect of carvacrol on lipopolysaccharide (LPS)-induced macrophage proinflammatory responses and endotoxic shock. The results showed that carvacrol significantly reduced mouse body weight loss and ameliorated pathological damage to the liver, lung, and heart under LPS-induced sepsis. Carvacrol attenuated inflammatory responses by inhibiting the LPS-induced production of inflammatory cytokine interleukin-6 (IL-6) in vivo and in vitro. Mechanistically, carvacrol inhibited IL-6 production mainly through the ERK1/2 signalling pathway in macrophages. Furthermore, carvacrol improved the survival of septic mice. This study sheds light on the role of carvacrol in the pathogenesis of LPS-induced sepsis, and thus, its potential in treating sepsis patients may be considered.


Bactericidal activity of tracheal antimicrobial peptide against respiratory pathogens of cattle.
Tracheal antimicrobial peptide (TAP) is a β-defensin produced by mucosal epithelial cells of cattle. Although effective against several human pathogens, the activity of this bovine peptide against the bacterial pathogens that cause bovine respiratory disease have not been reported. This study compared the antibacterial effects of synthetic TAP against Mannheimia haemolytica, Histophilus somni, Pasteurella multocida, and Mycoplasma bovis. Bactericidal activity against M. bovis was not detected. In contrast, the Pasteurellaceae bacteria showed similar levels of susceptibility to that of Escherichia coli, with 0.125μg TAP inhibiting growth in a radial diffusion assay and minimum inhibitory concentrations of 1.56-6.25μg/ml in a bactericidal assay. Significant differences among isolates were not observed. Sequencing of exon 2 of the TAP gene from 23 cattle revealed a prevalent non-synonymous single nucleotide polymorphism (SNP) A137G, encoding either serine or asparagine at residue 20 of the mature peptide. The functional effect of this SNP was tested against M. haemolytica using synthetic peptides. The bactericidal effect of the asparagine-containing peptide was consistently higher than the serine-containing peptide. Bactericidal activities were similar for an acapsular mutant of M. haemolytica compared to the wild type. These findings indicate that the Pasteurellaceae bacteria that cause bovine respiratory disease are susceptible to killing by bovine TAP and appear not to have evolved resistance, whereas M. bovis appears to be resistant. A non-synonymous SNP was identified in the coding region of the TAP gene, and the corresponding peptides vary in their bactericidal activity against M. haemolytica.


CO2 adsorption enhancement over Al/C-doped h-BN: A DFT study.
Reducing energy barriers of CO2 being chemisorbed on hexagonal boron nitride (h-BN) is a kernel step to efficiently and massively capture CO2. In this study, aluminum/carbon (Al/C) atoms are used as dopants to alter surface potential fields of h-BN, which aims at lowering energy barriers of adsorption processes. Through theoretical calculations, direct-adsorption structures/properties of CO2, joint-adsorption structures/properties of CO2/H2O, transition state (TS) energy barriers, effects of temperatures on adsorption energies/TS energy barriers and changes of reaction rate constants over different adsorbents are investigated in detail in order to reveal how doping of Al/C atoms promotes CO2 adsorption strength over doped h-BN. According to DFT calculation results, the average adsorption energy of CO2 being directly adsorbed on Al/C-doped h-BN arrives at -59.43 kJ/mol, which is about 5 times as big as that over pure h-BN. As to the average adsorption energy of CO2/H2O and relevant TS energy barrier, they are modified to -118.89 kJ/mol and 40.23 kJ/mol over Al/C-doped h-BN in contrast with -33.91 kJ/mol and 1695.11 kJ/mol over pure h-BN, respectively. What is more, based on thermodynamic analyses and reaction dynamics, the average desorption temperatures of CO2(/H2O) are promoted over doped h-BN and the temperature power exponent is negatively correlated with the activation energy in the Arrhenius equation form. The complete understanding of this study would supply crucial information for applying Al/C-doped h-BN to effectively capturing CO2 in real industries.


The use of digital rheoplethysmography for the study of psychogenic and neurogenic factors in man.
The use of the RPG in the qualitative and quantitative study of the state of the intact digital vasculature of intact man is illustrated and emphasized. Rheoplethysmography is the only method for quantitatively recording continuously and simultaneously the volumes and rates of inflow, outflow and the differences between inflow and outflow and the rate of basal blood flow in the digits of intact man or in any organ or tissue that can be isolated in a plethysmographic cup. The effects of psychogenic and neurogenic factors on the peripheral circulation of man were studied with the RPG, and the results demonstrate how useful this method is for such studies of the fingertip.


Poor results with percutaneous endoscopic jejunostomy.
A percutaneous endoscopic gastrostomy was placed in 20 malnourished patients to serve as a conduit for passage of a percutaneous endoscopic jejunostomy (PEJ) catheter for delivery of alimentation directly into the small bowel. Serious complications occurred in 95% of the patients and 50% of the subjects died. Aspiration was the most common adverse event and accounted for all deaths. Ten of 15 subjects (67%) treated with a PEJ to prevent aspiration continued to aspirate after the catheter was placed. PEJ tube failures were documented in 14 subjects (70%) and occurred because of occlusion, leakage, malposition, extrusion, cracking, kinking, or rupture of the catheter. These problems rendered the PEJ nonfunctional 18% of the time. Large manpower and resource investments were required to manage the PEJ and its complications. Our results suggest that enteral feeding through a PEJ does not prevent aspiration. Serious PEJ-related morbidity (95%), mortality (50%), and catheter failures (70%) occur. Refinements in methodology and catheter design will be required before additional use of this technique can be recommended.


Interactions between lysergic acid diethylamide and dopamine-sensitive adenylate cyclase systems in rat brain.
Investigations were carried out on the interactions of the hallucinogenic drug, D-lysergic acid diethylamide (D-LSD), and other serotonin antagonists with catecholamine-sensitive adenylate cyclase systems in cell-free preparations from different regions of rat brain. In equimolar concentration, D-LSD, 2-brono-D-lysergic acid diethylamide (BOL), or methysergide (UML) strongly blocked maximal stimulation of adenylate cyclase activity by either norepinephrine or dopamine in particulate preparations from cerebral cortices of young adult rats. D-LSD also eliminated the stimulation of adenylate cyclase activity of equimolar concentrations of norepinephrine or dopamine in particulate preparations from rat hippocampus. The effects of this hallucinogenic agent on adenylate cyclase activity were most striking in particulate preparations from corpus striatum. Thus, in 10 muM concentration, D-LSD not only completely eradicated the response to 10 muM dopamine in these preparations but also consistently stimulated adenylate cyclase activity. L-LSD (80 muM) was without effect. Significant activation of striatal adenylate cyclase was produced by 0.1 muM D-LSD. Activation of striatal adenylate cyclase of either D-LSD or dopamine was strongly blocked by the dopamine-blocking agents trifluoperazine, thioridazine, chlorpromazine, and haloperidol. The stimulatory effects of D-LSD and dopamine were also inhibited by the serotonin-blocking agents, BOL, 1-methyl-D-lysergic acid diethylamide (MLD), and cyproheptadine, but not by the beta-adrenergic-blocking agent, propranolol. However, these serotonin antagonists by themselves were incapable of stimulating adenylate cyclase activity in the striatal preparations. Several other hallucinogens, which were structurally related to serotonin, were also inactive in this regard, e.g., mescaline, N,N-dimethyltryptamine, psilocin and bufotenine. Serotonin itself produced a small stimulation of adenylate cyclase activity in striatal preparations and, in relatively high concentration (100 muM), partially blocked the activation by 10 muM dopamine, but was without effect on the stimulation by 10 muM D-LSD. The present results indicate that serotonin antagonists, in general, are potent inhibitors of catecholamine-induced stimulation of adenylate cyclase systems in brain cell-free preparations. In addition, these results, coupled with earlier findings on the capacity of D-LSD to interact with serotonin-sensitive adenylate cyclase systems from rat brain23,24 and other neural systems16, strongly suggest that this hallucinogenic agent is capable of acting as an agonist at central dopamine and serotonin receptors, as well as functioning as an antagonist at dopamine, norepinephrine, and serotonin receptors in the brain.


Signalling between the sexes during pollen tube reception.
Plant reproduction is a complex, highly-coordinated process in which a single, male germ cell grows through the maternal reproductive tissues to reach and fertilise the egg cell. Focussing on Arabidopsis thaliana, we review signalling between male and female partners which is important throughout the pollen tube journey, especially during pollen tube reception at the ovule. Numerous receptor kinases and their coreceptors are implicated in signal perception in both the pollen tube and synergid cells at the ovule entrance, and several specific peptide and carbohydrate ligands for these receptors have recently been identified. Clarifying the interplay between these signals and the downstream responses they instigate presents a challenge for future research and may help to illuminate broader principles of plant cell-cell communication.


Obsessive-compulsive tendencies are associated with a focused information processing strategy.
The study examined the hypothesis that obsessive-compulsive (OC) tendencies are related to a reliance on focused and serial rather than a parallel, speed-oriented information processing style. Ten students with high OC tendencies and 10 students with low OC tendencies performed the flanker task, in which they were required to quickly classify a briefly presented target letter (S or H) that was flanked by compatible (e.g., SSSSS) or incompatible (e.g., HHSHH) noise letters. Participants received 4 blocks of 100 trials each, two with 50% compatible trials and two with 80% compatible trials and were informed of the probability of compatible trials before the beginning of each block. As predicted, high OC participants, as compared to low OC participants, had slower overall reaction time (RT) and lower tendency for parallel processing (defined as incompatible trials RT minus compatible trials RT). Low, more than high OC participants tended to adjust their focused/parallel processing including a shift towards parallel processing in blocks with 80% compatible trials and in trials following compatible trials. Implications of these results to the cognitive theory and therapy of OCD are discussed.


Long-term performance, microbial evolution and spatial microstructural characteristics of anammox granules in an upflow blanket filter (UBF) treating high-strength nitrogen wastewater.
Granule formation, microstructure and microbial spatial distribution are crucial to granule stability and nitrogen removal. Here, an upflow blanket filter (UBF) reactor with porous fixed cylinder carriers was fabricated and operated for 234 days to investigate overall performance and the formation mechanism of anammox granules. Results showed that the UBF performed the highest nitrogen removal efficiency of 93.19 ± 3.39% under nitrogen loading rate of 3.6 kg-N/m3/d and HRT of 2 h. The tryptophan-like proteins as the key component in EPS were vital for granules formation. Further 16 s rRNA analysis indicated that SBR1031 with a relative abundance of 40.5% played an important role in cell aggregation. Thus, anammox granules were developed successfully with a two-layered spatial structure where outer-layer was ammonia oxidizing bacteria and inner-core was anaerobic ammonia oxidizing bacteria. Together, introduction of porous fixed cylinder carriers is a valid method to avoid biomass loss and floatation.


Using a point reduction clamp with a rubber stopper from a sterile bottle for reduction of a distal radius fracture.
There are many choices of surgical treatment for a distal radius fracture. The goal of treatment in these injuries is stable anatomical reduction of the articular surface. In a coronal split fracture, the dorsal fragment tends to dorsal displacement during drilling or when applying the distal locking screws of the plate. We present an illustrative case from a larger series of a 65-year-old Thai woman with an intraarticular distal radius fracture with a dorsal fragment from a coronal-split configuration reduced and stabilized with a volar locking plate utilizing a large point reduction clamp held in place with a rubber stopper from a sterile glass bottle to counter the displacement effect of the drilling. Using a large point reduction clamp with a rubber stopper from a sterile glass bottle enables this type of difficult fracture to be both reduced and stabilized with the locking screw easily inserted to stabilize the dorsal fragment without any further displacement. The rubber stopper acts to distribute the compressive force from the large point reduction clamp over a larger area allowing a more stable fracture stabilization, while at the same time reducing skin and soft tissue trauma at the dorsal aspect of the wrist. This workaround allows improved stability in reduction and stabilization of a coronal split intraarticular distal radius fracture. The advantage of this workaround is that it uses small things readily available in every operating room setting, and it does not require any special experience or skills.


Spatial and temporal trends in occurrence of emerging and legacy contaminants in the Lower Columbia River 2008-2010.
The Lower Columbia River in Oregon and Washington, USA, is an important resource for aquatic and terrestrial organisms, agriculture, and commerce. An 86-mile stretch of the river was sampled over a 3year period in order to determine the spatial and temporal trends in the occurrence and concentration of water-borne organic contaminants. Sampling occurred at 10 sites along this stretch and at 1 site on the Willamette River using the semipermeable membrane device (SPMD) and the polar organic chemical integrative sampler (POCIS) passive samplers. Contaminant profiles followed the predicted trends of lower numbers of detections and associated concentrations in the rural areas to higher numbers and concentrations at the more urbanized sites. Industrial chemicals, plasticizers, and PAHs were present at the highest concentrations. Differences in concentrations between sampling periods were related to the amount of rainfall during the sampling period. In general, water concentrations of wastewater-related contaminants decreased and concentrations of legacy contaminants slightly increased with increasing rainfall amounts.


Controversies in cardiopulmonary resuscitation.
This article reviews cardiopulmonary and cerebral resuscitation. New experimental and clinical information is presented, with some guidance for present therapy and future investigations.


Vacuum-assisted closure of pleural empyema without classic open-window thoracostomy.
A 64-year-old man was diagnosed with complex empyema after a second course of palliative chemotherapy for metastatic lung cancer. Because of the poor general condition of the patient, the decision was made to proceed with vacuum-assisted closure (VAC) therapy of the empyema without Eloesser or Clagett open-window thoracostomy (OWT). Installation and changing of the VAC sponge were performed using the ALEXIS Wound Protector/Retractor (Applied Medical, Rancho Santa Margarita, CA), a flexible polymer membrane tube. After 10 days of VAC treatment, the pleural cavity was sterile and was closed with single stitches. Chemotherapy was resumed 1 week later.


In situ degradation: a new concept for system suitability tests in monographs of the European Pharmacopoeia.
Monographs of the European Pharmacopoeia describe in the LC-test for related substances usually a system suitability test in order to ensure the adequate separation of impurities. Since the reference substances required are often not available a recent approach to avoid this problem is the generation of the required impurity by 'in situ degradation' of the active principle. This paper describes some typical applications of this technique as well as recent examples, such as the controlled degradation of cefalotin sodium, imipenem and spiramycin.


Does exercise training improve exercise tolerance, quality of life, and echocardiographic parameters in patients with heart failure with preserved ejection fraction? A systematic review and meta-analysis of randomized controlled trials.
Patients with heart failure with preserved ejection fraction (HFpEF) often experience dyspnea, decreased exercise tolerance, and decreased quality of life (QOL). Exercise training is a promising non-pharmacological treatment, with some improvement in exercise tolerance and QOL in HFpEF patients in randomized controlled trials (RCTs). We conducted a systematic review and meta-analysis of RCTs examining the effect of exercise therapy on exercise tolerance, QOL, and echocardiographic parameters in patients with HFpEF. Article database search of PubMed, Embase, and Cochrane Central Register of Controlled Trials identified 15 publications representing 579 unique patients. Results are presented as weighted mean difference (WMD) with 95% confidence intervals (CI). Exercise training (compared to control) demonstrated a significant improvement in exercise tolerance as measured by peak absolute VO2 (WMD [95% CI] = 164.67 [65.54, 263.79] mL/min), peak relative VO2 (WMD [95% CI] = 1.85 [0.98, 2.73] mL/min/kg), workload (WMD [95% CI] = 12.92 [4.67, 21.17] W), exercise time (WMD [95% CI] = 2.05 [1.57, 2.53] min), anaerobic threshold (WMD [95% CI] = 170.31 [35.40, 305.22] mL/min/kg), and 6-min walk test distance (WMD [95% CI] = 32.77 [20.72, 44.83] m); in QOL as measured by Short Form (SF-36) physical functioning domain (WMD [95% CI] = 9.95 [2.85, 17.05]) and SF-36 vitality domain (WMD [95% CI] = 6.24 [0.15, 12.34]); and in the echocardiographic measure of LVESD (WMD [95% CI] = - 0.16 [- 0.28, - 0.04] cm). In conclusion, we found after systematic review and meta-analysis of RCTs that exercise therapy improves exercise tolerance and physical-related quality of life measures.


Rheumatoid arthritis patients initiating rituximab with low number of previous bDMARDs failures may effectively reduce rituximab dose and experience fewer serious adverse events than patients on full dose: a 5-year cohort study.
Rituximab is used for the treatment of active rheumatoid arthritis. In the present study, we examined the long-term flare risk and safety of reduced doses of rituximab. This was a prospective, observational, single-center study of patients starting rituximab on standard dose (SD). Patients were switched to low dose (LD) (1 g every 6 months), based on the treating rheumatologist's decision after having achieved sustained clinical responses, while the rest of the patients continued on standard dose (SD). During a 60-month period, we assessed (Kaplan-Meier survival analysis) the relapse rate (increase ≥ 1.2 in DAS28-ESR for ≥ 6 months) and discontinuations due to treatment failure in the low dose group, and we compared the incidence of serious adverse events (SAEs) between LD and SD groups. Out of 361 patients [females 83.4%, mean age 61.9 (10.6) years, seropositive 50.3%, median total comorbidities count 4], 81 patients (22.4%) entered LD in a median time of 24 months (95% CI 18-30 months). Seropositivity (OR 1.823), more than 2 previous bDMARDs failures (OR 0.428), and DAS28 < 4.88 at 6 months (OR 2.329) predicted the odds of entering LD (p < 0.05 for all). During 60 months of follow-up, only 7.5% of patients on LD relapsed. Patients on LD had significantly less SAEs and all-cause hospitalizations as compared to the SD group (p < 0.05 for all). Linear regression analysis showed that previous hospitalization while on bDMARDs (p < 0.0001), use of prednisolone > 5 mg/day while on rituximab (p < 0.0001), and a history of ≥ 2 previous csDMARDs (p = 0.041) predicted the risk of SAEs. In a cohort of patients with established RA and significant comorbidities who taper rituximab after substantial initial disease activity improvement, a low rate of relapses and lower risk of SAEs compared to SD were recorded. Seropositivity, a lower number of previous bDMARDs use, and lower DAS28 at 6 months predicted the probability of entering the LD regimen.


Glucose tolerance in Polynesia: association with obesity and island of residence.
Data on five Polynesian populations, obtained by standardized population surveys conducted during the years 1978-1980, were examined for associations between glucose tolerance and both obesity and island of residence. In both sexes, after allowing for the influence of age and obesity, there was a significant difference in glucose tolerance between the three populations considered, subjectively, to be the less traditional and the two considered as retaining a more traditional lifestyle. Regression models predicting diabetic status were weaker than those using glucose tolerance as the dependent variable, probably due to the small number of diabetic subjects in the samples. As all subjects were of Polynesian ancestry, and the results could not be explained by knowledge of ancestral affiliations between the five populations, environmental, rather than genetic factors may have been the determinants of the observed differences in glucose tolerance. This finding highlights the need for a more sophisticated approach to the study of the association between socio-cultural modernization and chronic disease in the Pacific.


The effects of erdosteine, N-acetylcysteine, and vitamin E on nicotine-induced apoptosis of hippocampal neural cells.
This study investigated the frequency of apoptosis in rat hippocampal neural cells after intraperitoneal nicotine injection, examining the roles of the inflammatory markers myeloperoxidase (MPO) and tumor necrosis factor alpha (TNF-alpha) in nicotine-induced brain damage and the protective effects of three known antioxidant agents, N-acetylcysteine (NAC), erdosteine, and vitamin E. Female Wistar rats were divided into seven groups, each composed of nine rats: 2 negative control groups, 2 positive control groups, one erdosteine-treated group (500 mg/kg), one NAC-treated group (500 mg/kg), and one vitamin E-treated group (500 mg/kg). Nicotine was intraperitoneally injected at a dosage of 0.6 mg/kg for 21 days. Following nicotine injection, the antioxidants were administered orally; treatment was continued until the rats were killed. Apoptosis level in hippocampal neural cells was determined by using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling) method. Staining of cytoplasmic TNF-alpha in hippocampal neural cells and hippocampus MPO activity were evaluated by immunohistochemistry. Nicotine administration had no effect on local TNF-alpha production, or hippocampal MPO activity. The treatments with erdosteine, NAC and vitamin E significantly reduced the rate of nicotine-induced hippocampal neural cell apoptosis. This findings suggest that erdosteine and NAC can be as effective as vitamin E in protecting against nicotine-induced hippocampal neural cell apoptosis.


Insights into predicting diabetic nephropathy using urinary biomarkers.
Diabetic nephropathy (DN) is a serious complication of diabetes caused by changes in the structure and function of the kidneys. It is important to detect diagnostic biomarkers of DN at an early stage, in which the drug can slow the loss of kidney function and prevent disease progression. In recent years, a variety of biological markers related to DN have been discovered, which is of great significance for predicting the occurrence and development of diseases. Due to the simplicity of non-invasive collection, urine is an ideal biological sample for the discovery of new biomarkers of kidney disease. We reviewed some new urinary biomarkers related to early DN patients, including urinary proteins, peptides, and exosomes biomarkers. We also highlight the proteins associated with tubular damage, glomerular damage, inflammation and oxidative stress marker. Despite the promise of these new urinary biomarkers, we next proposed a review of the most recent publications reporting on larger cohorts, focusing on those that aim at qualification or validation. This review provides important data to better understand biomarkers related to the pathophysiology of DN, and these markers have been increasingly studied for disease progression to provide effective human treatment.


Thermal entrainment methods for studying Raynaud's phenomenon.
Entrainment occurs when an externally applied periodic temperature stimulus forces the peripheral bloodflow component of thermoregulation to oscillate at the same frequency. This phenomenon can be demonstrated using frequency transforms to analyse the spectral content of the bloodflow, and can be used as a diagnostic test for Raynaud's phenomenon. Correlations were performed between the clinical diagnosis and the objective tests. The average inter-clinical correlation coefficient was r = 0.66. When the average clinical diagnosis was compared with a combination of thermal entrainment and digital patency testing the correlation coefficient rose to r = 0.68. These results highlight the difficulties encountered when assessing patients with Raynaud's phenomenon and the necessity of applying both physiological and clinical techniques.


Metachronous benign ovarian tumors are not uncommon in children.
To evaluate the risk for metachronous ovarian tumor in pediatric patients with mature ovarian teratoma. During 1981-2011, 22 children underwent oophorectomy for mature teratoma at the median age of 11.4 (range 1.5-15.3) years. The patients were followed-up in median 4.4 (range 0.5-25.5) years. None of the patients had synchronous bilateral tumor at the time of primary operation, but during follow-up five patients (23%) got metachronous contralateral ovarian tumor. The contralateral tumor was observed in median 3.6 (range 1-8.8) years after the primary operation. According to Kaplan-Meier analysis the risk for contralateral tumor was 14% ± 8% (SE) within five years and 66% ± 26% (SE) within 10 years. In this series, the contralateral tumor was operated by ovary preserving surgery. Three of the metachronous tumors were mature teratomas and two were seromucinous infantile cystadenomas. One patient had a second teratoma recurrence 14 years after the first recurrence. More than one fifth of the children with ovarian mature teratoma get metachronous benign tumor to the contralateral ovary. Therefore a yearly ultrasound follow-up is needed for these patients up to potential pregnancy to enable early diagnosis, ovary preserving surgery and maintenance of fertility in the case of metachronous tumor.


Arteriogenesis in the pia matter of the rat brain cortex after intracerebral injection of mesenchymal stem cells.
We studied the effect of intracerebral transplantation of mesenchymal stem cells on the density of pial arterioles in rat brain cortex. It was shown that intracerebral transplantation although causes damage to about half the area of the ipsilateral pia mater, does not change the density of the microvascular network neither in border region of the injured tissue, nor in the contralateral hemisphere. Intracerebral transplantation of mesenchymal stem cells promoted arteriogenesis in the pia mater of the contralateral hemisphere: density of arterioles in this area was significantly (by about 2.5 times higher) than in other experimental animals.


Catalytic properties and chemical composition of pepsins from Atlantic cod (Gadus morhua).
1. Three pepsins were purified from the gastric mucosa of Atlantic cod (Gadus morhua). 2. The enzymes, called Pepsin I and Pepsin IIa and b, had isoelectric points 6.9, 4.0 and 4.1, respectively, and digested hemoglobin at a maximal rate at a pH of approximately 3. 3. They resembled bovine cathepsin D in being unable to digest the mammalian pepsin substrate N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine. 4. Specificity constants (kcat/Km) for the cod pepsins were lower than for porcine pepsin, and they expressed higher substrate affinity and physiological efficiency at pH 3.5 than at pH 2. 5. The cod pepsins are glycoproteins, and their amino acid composition resembles that of porcine cathepsin D more than that of porcine pepsin. 6. The N-terminal sequence of Atlantic cod pepsins is substantially different from that of porcine pepsin. This indicates a significant evolutionary gap between fish and mammalian pepsins.


Microvascular surgery as an adjunct to craniomaxillofacial reconstruction.
Fifteen patients (8 female and 7 male) age 4 to 57 years underwent microsurgical free tissue transfers as a component of a craniomaxillofacial reconstruction. In 12 patients the free flap was performed simultaneously with the bony procedure and in three it was a secondary procedure. The patients included two craniosynostosis, four craniofacial tumours, five hemicraniofacial microsomias, one facial and skull base arteriovenous malformation, one orbitofacial neurofibromatosis and two hemifacial atrophies with extensive facial skeletal involvement. The rectus abdominis free flap was used in 9 patients, the latissimus dorsi in two, the omentum in three, and the first web-space in one. The choice of tissue varied according to the size of the defect and its location. The rectus abdominis musculofasciocutaneous flap was the most frequent source of tissue for contour restoration, and the omentum was used to fill intracranial spaces. One flap failed intraoperatively in a patient with hemifacial microsomia and inadequate and abnormal recipient vessels. One patient had an injury to the temporal branch of the facial nerve, with spontaneous recovery. In 13 patients the free tissue transfer was for soft tissue fill with cover of facial bone or skull base; secondary procedures were frequently required in these patients. In two patients with intracranial free flaps, no further procedures were necessary. In selected cases the association of microvascular techniques with craniomaxillofacial surgery can facilitate reconstruction and improve results.


Histological analysis of indirect somatic embryogenesis in the Marsh clubmoss Lycopodiella inundata (L.) Holub (Pteridophytes).
An efficient in vitro plant regeneration method was developed for Lycopodiella inundata (L.) Holub, an endangered medicinal Lycopod (Pteridophytes). Vegetative apices were used as explant material. Nodular calluses were established after three cycles (13 weeks each) on a medium containing a few minerals and organic compounds and supplemented with 0.05 µM 3-indolebutyric acid (IBA) and 1.4 µM kinetin (Kin). Propagation was achieved every 13 weeks on this callus medium (CM). When nodular calluses were transferred on a medium supplemented with 2.5 µM IBA and 0.33 µM gibberellic acid (GA(3)) designated as embryogenic medium (EM), organized structures appeared and developed into plantlets. Development phases were characterized by histological studies. Some phases of zygotic embryogenesis previously described for Lycopods were observed in L. inundata. Histological analyses established that an indirect somatic embryo was derived from a single embryogenic cell by following the zygotic developmental pathway. As this phenomenon has not previously been reported in Lycopods, a comparison between somatic and zygotic embryos is discussed based upon morphology and histology.


Fucoxanthin Alleviates Lipopolysaccharide-Induced Inflammation and Immunosuppression in RAW264.7 Macrophages.
Fucoxanthin (FX) has been reported to reduce mortality in mouse models of sepsis, but its exact cause remains to be determined. In the present study, we evaluated the immunomodulatory properties of FX in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our results showed that FX could not only suppress the immune activation responses caused by LPS primary stimulation, but also antagonize LPS restimulation-induced immunosuppression in macrophages. The immunomodulatory capabilities of FX was mainly demonstrated by regulating the production of the inflammatory mediator under different LPS stimuli. Furthermore, we found that adenosine monophosphate-activated protein kinase (AMPK) activation was required for FX's anti-inflammatory and anti-immunosuppressive activities. Our results complement existing data supporting the clinical potential for FX in treating sepsis.


A multiomics approach-based prediction of radiation pneumonia in lung cancer patients: impact on survival outcome.
To predict the risk of radiation pneumonitis (RP), a multiomics model was built to stratify lung cancer patients. Our study also investigated the impact of RP on survival. This study retrospectively collected 100 RP and 99 matched non-RP lung cancer patients treated with radiotherapy from two independent centres. They were divided into training (n = 175) and validation cohorts (n = 24). The radiomics, dosiomics and clinical features were extracted from planning CT and electronic medical records and were analysed by LASSO Cox regression. A multiomics prediction model was developed by the optimal algorithm. Overall survival (OS) between the RP, non-RP, mild RP, and severe RP groups was analysed by the Kaplan‒Meier method. Sixteen radiomics features, two dosiomics features, and one clinical feature were selected to build the best multiomics model. The optimal performance for predicting RP was the area under the receiver operating characteristic curve (AUC) of the testing set (0.94) and validation set (0.92). The RP patients were divided into mild (≤ 2 grade) and severe (> 2 grade) RP groups. The median OS was 31 months for the non-RP group compared with 49 months for the RP group (HR = 0.53, p = 0.0022). Among the RP subgroup, the median OS was 57 months for the mild RP group and 25 months for the severe RP group (HR = 3.72, p < 0.0001). The multiomics model contributed to improving the accuracy of RP prediction. Compared with the non-RP patients, the RP patients displayed longer OS, especially the mild RP patients.


Diagnostic significance of blood eosinophil count in eosinophilic chronic rhinosinusitis with nasal polyps in Chinese adults.
Over half of chronic rhinosinusitis with nasal polyps (CRSwNP) patients in China show noneosinophilic inflammation. The aim of this study was to compare the clinical characteristics of eosinophilic and noneosinophilic CRSwNP and to identify the predictors of eosinophilic CRSwNP. A retrospective study. There were 155 CRSwNP patients enrolled in the Tongji cohort. Eosinophilic CRSwNP was diagnosed according to our previously published histologic criterion. The demographic and clinical features were compared between eosinophilic and noneosinophilic CRSwNP. Factors associated with eosinophilic CRSwNP were determined with regression analysis, and optimal cutoff points of the predictors were determined by a receiver operating characteristic curve. The optimal cutoff points of the predictors were validated in an independent group of 35 CRSwNP patients referred to as the Taizhou cohort. A male preponderance, a higher prevalence of smoking and atopy, and higher peripheral blood eosinophil absolute count and percentage and blood IgE levels were found in eosinophilic CRSwNP compared with noneosinophilic CRSwNP. Peripheral eosinophil absolute count and percentage were independently and significantly associated with eosinophilic CRSwNP. An absolute blood eosinophil count ≥ 0.215 × 10(9) /L yielded a sensitivity of 74.2% and a specificity of 86.5%, and a blood eosinophil percentage ≥ 3.05% yielded a sensitivity of 80.3% and a specificity of 75.3% for the diagnosis of eosinophilic CRSwNP in the Tongji cohort. The validation study in the Taizhou cohort revealed a lower sensitivity and specificity. Eosinophilic and noneosinophilic CRSwNP displayed significant differences in certain clinical features. Peripheral blood eosinophil count could distinguish eosinophilic and noneosinophilic CRSwNP in Chinese adults.


Improved tensile strength of glycerol-plasticized gluten bioplastic containing hydrophobic liquids.
The aim of the present work has been to study the influence of hydrophobic liquids on the morphology and the properties of thermo-molded plastics based on glycerol-plasticized wheat gluten (WG). While the total amount of castor oil and glycerol was remained constant at 30 wt%, castor oil with various proportions with respect to glycerol was incorporated with WG by mixing at room temperature and the resultant mixtures were thermo-molded at 120 degrees C to prepare sheet samples. Moisture absorption, morphology, dynamic mechanical properties, and tensile properties (Young's modulus, tensile strength and elongation at break) of the plastics were evaluated. Experimental results showed that the physical properties of WG plastic were closely related to glycerol to castor oil ratio. Increasing in castor oil content reduces the moisture absorption markedly, which is accompanied with a significant improvement in tensile strength and Young's modulus. These observations were further confirmed in 24 wt% glycerol-plasticized WG plastics containing 6 wt% silicone oil or polydimethylsiloxane (PDMS) liquid rubber.


Assessment of Ischemic Volumes by Using Relative Filling Time Delay on CTP Source Image in Patients with Acute Stroke with Anterior Circulation Large Vessel Occlusions.
Thrombectomy up to 24 hours after stroke onset in patients with specific ischemic brain volumes remains a challenge, because many stroke centers do not apply specialized software to calculate ischemic volumes at advanced imaging. We aimed to establish the association between relative filling time delay on CTP source imaging and ischemic volume parameters and the infarct penumbra to core volume mismatch in patients with acute ischemic stroke. Consecutive patients with acute ischemic stroke and with M1 segment MCA with or without terminal ICA occlusions on baseline CTA and CTP within 24 hours of stroke symptom onset were included. Ischemic volumes were analyzed with software based on CTP maps. Relative filling time delay was classified into 4 grades-grade 0: relative filling time delay = 0 seconds; grade 1: relative filling time delay >0 to ≤4 seconds; grade 2: relative filling time delay >4 to ≤8 seconds; and grade 3: relative filling time delay > 8 seconds. Differences in ischemic volume parameters among relative filling time delay grades were tested. We recruited 138 patients (median age, 69 years; 62.3% male). Different median volumes of the infarct core (grade 0, 7.3 mL; grade 1, 23.3 mL; grade 2, 45.7 mL; grade 3, 135 mL [P < .001]) and the penumbra (grade 0, 47.6 mL; grade 1, 90 mL; grade 2, 110 mL; grade 3, 92 mL [P = .043]) were observed among relative filling time delay grades. Target mismatch (defined by the criteria of the DEFUSE 3 trial) was identified in 71.7% of the patients (99/138). A relative filling time delay grade ≤ 1 independently predicted target mismatch, with a sensitivity of 0.79 (95% CI, 0.7-0.87) and a specificity of 0.66 (95% CI, 0.49-0.8). Relative filling time delay grade based on CTP source imaging is a simple and effective parameter for evaluating ischemic volumes and target mismatch in patients with acute ischemic stroke. Further studies that compare relative filling time delay grade with clinical functional outcomes are necessary.


Monozygous twin brothers discordant for photosensitive epilepsy: first report of possible visual priming in humans.
The interaction of genetic predisposition and the environment in the development of epilepsy is often discussed, but, aside from some animal reflex epilepsies, little evidence supports such interaction in the development of reflex epilepsy in humans. We describe the history of a 16-year-old boy in whom photosensitive epilepsy developed after a period of weekly exposures to high-intensity light flashes. Both he and his clinically unaffected monozygotic twin were found to be photosensitive. This case report suggests that some genetic forms of human reflex epilepsy may be elicited by repeated environmental exposure to the appropriate stimulus, similar to some of the stimulus-induced epilepsies seen in animals.


Differences in the association between persistent pathogens and mood disorders among young- to middle-aged women and men in the U.S.
A growing literature supports the role of immune system alterations in the etiology of mood regulation, yet there is little population-based evidence regarding the association between persistent pathogens, inflammation and mood disorders among younger women and men in the U.S. We used data from the National Health and Nutrition Examination Survey III on individuals 15-39 years of age assessed for major depression, dysthymia, and/or bipolar disorder I and tested for cytomegalovirus (N=6825), herpes simplex virus (HSV)-1 (N=5618) and/or Helicobacter pylori (H. pylori) (N=3167) seropositivity as well as C-reactive protein (CRP) level (N=6788). CMV immunoglobulin G (IgG) antibody level was also available for a subset of women (N=3358). We utilized logistic regression to estimate the odds ratio (OR) and 95% confidence interval (CI) for the association between pathogens, CRP levels and each mood disorder overall and among women and men, separately. H. pylori seropositivity was associated with increased odds of dysthymia (OR 2.37, 95% confidence interval (CI): 1.07, 5.24) among women, but decreased odds among men (OR 0.51, 95% CI: 0.28, 0.92). CMV seropositivity was also associated with lower odds of depression (OR 0.54, 95% CI: 0.32, 0.91) among men, while elevated CMV IgG level was marginally associated with increased odds of mood disorders among women. Associations were not mediated by CRP level. Our findings suggest that persistent pathogens such as CMV and H. pylori may differentially influence mood disorders among women and men, warranting further investigation into biological and/or sociocultural explanations for the contrasting associations observed.


Identification of limonoids from Walsura yunnanensis and evaluation of their cytotoxicity against cancer cell lines.
Three new limonoids, walsurauias A-C (1-3), along with four known ones, were isolated from the leaves and twigs of Walsura yunnanensis C. Y. Wu. Their structures were determined on the basis of comprehensive spectroscopic data analysis. The new limonoids were screened for their cytotoxic activity (IC50 0.81-5.73 μM) against four human cancer cell lines, including A549, HepG2, HCT116 p21KO and CNE-2. And α,β-unsaturated ketone moieties in rings A and B are essential for their cytotoxic activity. Selected compounds were further investigated. Compounds 1-3 effectively induced G2/M cell cycle arrest and apoptosis in a dose-dependent manner in cancer cells. In addition, compounds 1-3 inhibited the colony formation and compounds 2 and 3 suppressed the migration of cancer cells.


Insulin resistance in moderate chronic heart failure is related to hyperleptinaemia, but not to norepinephrine or TNF-alpha.
Chronic heart failure (CHF) has emerged as an insulin-resistant state, independently of ischaemic aetiology. The underlying mechanisms of this finding are not known. Catecholamines, tumor necrosis factor alpha (TNFalpha) and leptin, the adipocyte specific hormone, have all been implicated as mediators of impaired insulin sensitivity. The purpose of this study was to examine in patients with CHF and in comparison to healthy controls subjects whether norepinephrine, TNFalpha or leptin relate to insulin sensitivity. 41 patients with CHF (age 60+/-2 years, NYHA I/II/III/IV 4/12/22/3, peak oxygen consumption 17.6+/-1.0 ml/kg per min) and 21 healthy controls of similar age and total and regional fat distribution were studied in a cross-sectional study. Insulin sensitivity was assessed by intravenous glucose tolerance testing using the minimal model approach; catecholamines, TNFalpha and soluble TNF receptors 1 and 2 were also measured. Total and regional body fat mass was assessed by dual energy X-ray absorptiometry. Insulin sensitivity was reduced in CHF patients compared to controls by 31% (P<0.01) and fasting insulin was higher in patients than in controls (79.1+/-9.7 vs. 41.4+/-6.0 pmol/l, P<0.01). Patients had, compared to healthy controls, elevated serum leptin levels (8.28+/-0.84 vs. 4.83+/-0.68 ng/ml), norepinephrine (3.45+/-0.34 vs. 1.87+/-0.16 nmol/l, both P<0.01) and soluble TNF-receptors 1 (1280+/-141 vs. 639+/-52 pg/ml) and 2 (2605+/-184 vs. 1758+/-221 pg/ml, both P<0.01). Leptin levels corrected for total body fat mass were higher in CHF patients than in controls (41.3+/-3 vs. 24.3+/-2 pg/ml per 100 g, P<0.001). TNFalpha was not significantly different between the groups. In both groups there was an inverse correlation between insulin sensitivity and serum leptin (r=-0.65, P<0.0001 for pooled subjects); in contrast, no significant relation was found between insulin sensitivity and norepinephrine or TNFalpha. In multivariate regression analysis, leptin emerged as the only significant predictor of insulin sensitivity (standardised coefficient=-0.59, P<0.001), independent of body fat mass, age and peak VO2. In moderate CHF, elevated leptin levels directly and independently predict insulin resistance. Elevated serum leptin levels could play a role in the impaired regulation of energy metabolism in CHF. In contrast to observations in other conditions, TNFalpha and norepinephrine are not related to insulin resistance in moderate CHF.


Absence of carcinoembryonic antigen-like material in mesothelioma: an immunohistochemical differentiation from other lung cancers.
This study is to examine the potential usefulness of immunohistochemical staining for carcinoembryonic antigen (CEA)-like material in the differential diagnosis of mesotheliomas (12 cases) from other lung cancers (14 cases) that had been previously diagnosed by transmission and scanning electron microscopy and conventional light microscopy. Indirect immunofluorescent staining for CEA was carried out on formalin-fixed paraffin-embedded sections, and the slides were examined under code. All 9 cases of diffuse mesothelioma were negative, and all 12 cases of adenocarcinoma and bronchioloalveolar carcinoma were positive for CEA-like material. Three localized mesotheliomas and a carcinoid tumor were also negative. A squamous cell carcinoma was positive. A positive immunohistochemical result for CEA-like material in lung cancers will raise the possibility of its being of bronchial epithelial origin.


Two new species and additional records of Amphichroum Kraatz, 1857 from Nepal, China and India (Coleoptera: Staphylinidae: Omaliinae: Anthophagini).
Two new species of the genus Amphichroum Kraatz, 1857 are described and illustrated: A. brachypterum sp. n. (central Nepal) and A. cuccodoroi sp. n. (China: Yunnan). Faunistic records of A. altivagans Cameron, 1941 from India (Kashmir) and A. pindarense Champion, 1920 from Nepal are provided.


Ring-truncated deguelin derivatives as potent Hypoxia Inducible Factor-1α (HIF-1α) inhibitors.
A series of fluorophenyl and pyridine analogues of 1 and 2 were synthesized as ring-truncated deguelin surrogates and evaluated for their HIF-1α inhibition. Their structure-activity relationship was systematically investigated based on the variation of the linker B-region moiety. Among the inhibitors, compound 25 exhibited potent HIF-1α inhibition in a dose-dependent manner and significant antitumor activity in H1299 with less toxicity than deguelin. It also inhibited in vitro hypoxia-mediated angiogenic processes in HRMECs. The docking study indicates that 25 occupied the C-terminal ATP-binding pocket of HSP90 in a similar mode as 1, which implies that the anticancer and antiangiogenic activities of 25 are derived from HIF-1α destabilization by binding to the C-terminal ATP-binding site of hHSP90.


Non-obstructive low attenuation coronary plaque predicts three-year acute coronary syndrome events in patients with hypertension: multidetector computed tomographic study.
Arterial hypertension is an established risk factor for acute coronary syndrome (ACS). Multidetector computed tomography (MDCT) is an accurate and less invasive technique for assessment of the degree of coronary artery luminal narrowing and characterization of coronary atherosclerosis. We therefore aimed to investigate the predictive power of MDCT for future ACS events and compared with traditional parameters in patients with hypertension. One hundred and thirty-four patients (93 men, mean age 70±11 years) with hypertension underwent MDCT for evaluation of coronary artery disease. MDCT analysis focused on the presence of plaques, the degree of stenosis, and the plaque characteristics. Traditional parameters included Framingham risk score, carotid intima-media thickness, and left ventricular mass index. During a mean follow-up of 39±10 months, ACS events occurred in 10 patients, including myocardial infarction (n=3) and unstable angina (n=7). Multivariate analysis identified total number of low attenuation plaques as an independent predictor of ACS events (p<0.001). We demonstrated that non-obstructive low attenuation coronary plaques on MDCT predicted more accurately future ACS events in patients with hypertension than traditional parameters.


Transmission of the oral bacterium Streptococcus mutans to young Macaca fascicularis monkeys from human nursery attendants.
Streptococcus mutans was detected in the mouths of 2 out of 7 infant monkeys that had been delivered by caesarian section and nursery-reared without contact with their mothers or other monkeys. Their human nursery attendants all harboured Strep. mutans biotype c (c serotype) as found in the monkeys. Bacteriocin studies also indicated similarity between the animal and human isolates. These findings suggest that the infant monkeys acquired Strep. mutans from the nursery personnel. Evidence for the reverse transmission of enterococci, from monkeys to man was also obtained.


Herbal medicines for metabolic diseases with blood stasis: A protocol for a systematic review and meta-analysis.
Herbal medicines have the potential to be alternative treatments for metabolic diseases. This systematic review will assess the efficacy of herbal medicines in treating metabolic diseases with blood stasis. We will search MEDLINE, the Cochrane Central Register of Controlled Trials (CENTRAL), Web of Science, 2 Korean medical databases (OASIS, NDSL), a Chinese database (China National Knowledge Infrastructure, CNKI), and a Japanese database (J-STAGE) for relevant literature. We will include all randomized controlled trials (RCTs) or quasi-RCTs evaluating the effectiveness of herbal medicine. Participants of both sexes and of any age with clinically diagnosed metabolic disease with blood stasis will be included. Primary outcomes will include Blood-stasis syndrome score, TC, TG, HDL-C and LDL-C. Secondary outcomes will Blood pressure, FBS. ECG, prevalence rate of heart disease, and response rate. The risk of bias will be assessed using the Cochrane tool for assessing risk of bias. The findings of this study will provide a summary of the current state of evidence regarding the effectiveness of types of herbal medicine in managing metabolic disease with blood stasis. In addition, this review will be expected to provide a base for clinical trials to confirm the efficacy of herbal medicine for treatments of metabolic disease with blood stasis. PROSPERO 2018 CRD42018090179.


Hypotonicity induces TRPV4-mediated nociception in rat.
We hypothesized that TRPV4, a member of the transient receptor family of ion channels, functions as a sensory transducer for osmotic stimulus-induced nociception. We found that, as expected for a transducer molecule, TRPV4 protein is transported in sensory nerve distally toward the peripheral nerve endings. In vivo single-fiber recordings in rat showed that hypotonic solution activated 54% of C-fibers, an effect enhanced by the hyperalgesic inflammatory mediator prostaglandin E2. This osmotransduction causes nociception, since administration of a small osmotic stimulus into skin sensitized by PGE2 produced pain-related behavior. Antisense-induced decrease in expression of TRPV4 confirmed that the channel is required for hypotonic stimulus-induced nociception. Thus, we conclude that TRPV4 can function as an osmo-transducer in primary afferent nociceptive nerve fibers. Because this action is enhanced by an inflammatory mediator, TRPV4 may be important in pathological states and may be an attractive pharmacological target for the development of novel analgesics.


Proline-, glutamic acid-, and leucine-rich protein 1 mediates estrogen rapid signaling and neuroprotection in the brain.
17-β estradiol (E2) has been implicated as neuroprotective in a variety of neurodegenerative disorders. However, the underlying mechanism remains unknown. Here, we provide genetic evidence, using forebrain-specific knockout (FBKO) mice, that proline-, glutamic acid-, and leucine-rich protein 1 (PELP1), an estrogen receptor coregulator protein, is essential for the extranuclear signaling and neuroprotective actions of E2 in the hippocampal CA1 region after global cerebral ischemia (GCI). E2-mediated extranuclear signaling (including activation of extracellular signal-regulated kinase and Akt) and antiapoptotic effects [such as attenuation of JNK signaling and increase in phosphorylation of glycogen synthase kinase-3β (GSK3β)] after GCI were compromised in PELP1 FBKO mice. Mechanistic studies revealed that PELP1 interacts with GSK3β, E2 modulates interaction of PELP1 with GSK3β, and PELP1 is a novel substrate for GSK3β. RNA-seq analysis of control and PELP1 FBKO mice after ischemia demonstrated alterations in several genes related to inflammation, metabolism, and survival in PELP1 FBKO mice, as well as a significant reduction in the activation of the Wnt/β-catenin signaling pathway. In addition, PELP1 FBKO studies revealed that PELP1 is required for E2-mediated neuroprotection and for E2-mediated preservation of cognitive function after GCI. Collectively, our data provide the first direct in vivo evidence, to our knowledge, of an essential role for PELP1 in E2-mediated rapid extranuclear signaling, neuroprotection, and cognitive function in the brain.


Is dyslipidaemia important if we control glycaemia?
Cardiovascular disease is responsible for a large proportion of morbidity and mortality in people with type 2 diabetes, who typically have a clustering of risk factors for coronary heart disease (CHD), including dyslipidaemia. Intensive single-factor interventions to lower blood glucose have not eliminated the increased risk of adverse cardiovascular outcomes in patients with diabetes. In comparison, multifactorial interventions, which include normalizing blood lipids with statins, appear to be more effective in reducing CHD. However, risk reductions in patients with type 2 diabetes are often no better than those seen in people without diabetes, despite the much greater baseline risk in the former group. High-density lipoprotein cholesterol (HDL-C) is an independent risk factor for CHD, but is only minimally improved by statin therapy. Additional interventions aimed at increasing HDL-C can further reduce cardiovascular morbidity and mortality in patients with type 2 diabetes. However, use of fibrates in combination with statins may pose an unacceptable risk of myopathy. Therefore, other strategies for combination therapy are needed to normalize lipids in patients with type 2 diabetes or metabolic syndrome.