Small rubber particle proteins from Taraxacum brevicorniculatum promote stress tolerance and influence the size and distribution of lipid droplets and artificial poly(cis-1,4-isoprene) bodies.
Natural rubber biosynthesis occurs on rubber particles, i.e. organelles resembling small lipid droplets localized in the laticifers of latex-containing plant species, such as Hevea brasiliensis and Taraxacum brevicorniculatum. The latter expresses five small rubber particle protein (SRPP) isoforms named TbSRPP1-5, the most abundant proteins in rubber particles. These proteins maintain particle stability and are therefore necessary for rubber biosynthesis. TbSRPP1-5 were transiently expressed in Nicotiana benthamiana protoplasts and the proteins were found to be localized on lipid droplets and in the endoplasmic reticulum, with TbSRPP1 and TbSRPP3 also present in the cytosol. Bimolecular fluorescence complementation confirmed pairwise interactions between all proteins except TbSRPP2. The corresponding genes showed diverse expression profiles in young T. brevicorniculatum plants exposed to abiotic stress, and all except TbSRPP4 and TbSRPP5 were upregulated. Young Arabidopsis thaliana plants that overexpressed TbSRPP2 and TbSRPP3 tolerated drought stress better than wild-type plants. Furthermore, we used rubber particle extracts and standards to investigate the affinity of the TbSRPPs for different phospholipids, revealing a preference for negatively charged head groups and 18:2/16:0 fatty acid chains. This finding may explain the effect of TbSRPP3-5 on the dispersity of artificial poly(cis-1,4-isoprene) bodies and on the lipid droplet distribution we observed in N. benthamiana leaves. Our data provide insight into the assembly of TbSRPPs on rubber particles, their role in rubber particle structure, and the link between rubber biosynthesis and lipid droplet-associated stress responses, suggesting that SRPPs form the basis of evolutionarily conserved intracellular complexes in plants.


Antineoplastic activity of povidone-iodine on different mesothelioma cell lines: results of in vitro study.
Povidone-iodine (PVP-I) or Betadine, owing to its antineoplastic activity, is also used as an adjuvant during intra-abdominal or intrathoracic surgery. However, the protocol of PVP-I administration has not been optimized to achieve the best antitumoural efficacy. We aimed to determine the optimal concentration of PVP-I, the time of incubation and the mechanism of cell death by analysing the effect of different doses and time of administration of PVP-I on the cell viability of different mesothelioma cell lines. Four different cell lines (MET 5A/normal mesothelium; H2052/sarcomatoid mesothelioma; ISTMES2/epithelial mesothelioma; MSTO/biphasic mesothelioma) were incubated with increasing concentrations of diluted PVP-I (0.0001; 0.001; 0.01; 0.1; 1%) for 5, 10, 30, 60 min and 24 h, respectively. Cell viability was determined using cell direct cytotoxicity assay and cell death was determined through flow cytometry assay analysis. The superoxide dismutase activity was assessed functionally through a specific inhibitor to evaluate the mechanism of cell death. The antiproliferative effect of PVP-I varied largely among different cell lines in a dose- and time-dependent manner. At 0.1% concentration for 10 min of incubation, the percentage of viable cells was 0.5 ± 0.1; 0.8 ± 0.5 and 0% (P < 0.01) for MET5A, ISTMES2 and MSTO, respectively. Conversely, the same concentration did not significantly affect the H2052 cell line which was completely suppressed at a 1% concentration of PVP-I. Double staining of Annexin V and DNA showed that PVP-I induced cell death in all four cell lines via necrosis depending on PVP-I concentration. However, H2052 was found to be more resistant than MSTO, ISTMES2 and MET 5A cells lines. The activity of superoxide dismutase was significantly inhibited in all cell lines. Our results confirmed the anti-neoplastic activity of PVP-I especially on ISTMES2 and MSTO cell lines. With respect to chemotherapy pleural irrigation, washing with PVP-I is cost-effective and easy. If confirmed by larger studies, our findings suggest that the intrapleural irrigation with PVP-I (0.1% concentration for 10 min) in patients with epithelial or biphasic mesothelioma undergoing cytoreductive surgery might be applied in thoracic surgery practice to prevent neoplastic cell growth.


Antiviral treatment in patients with cytomegalovirus positive ulcerative colitis.
Cytomegalovirus (CMV) is a common virus in patients with ulcerative colitis receiving immunosuppressive drugs. Many studies suggested that CMV infection is an exacerbating factor in patients with ulcerative colitis. The role of CMV in exacerbations of ulcerative colitis has been discussed. One of studies starting this discussion is an article entitled "CMV positive ulcerative colitis: A single center experience and literature review" by Kopylov et al. However, we think that there are some points that should be emphasized about the study. Especially, the small number of patients in the study has led to meaningless results. Large controlled prospective trials are needed to clarify the benefit of antiviral therapy for active ulcerative colitis patients.


Progress and Clinical Applications of Crohn's Disease Exclusion Diet in Crohn's Disease.
Crohn's disease is a chronic intestinal inflammatory disorder of unknown etiology. Although the pharmacotherapies for Crohn's disease are constantly updating, nutritional support and adjuvant therapies have recently gained more attention. Due to advancements in clinical nutrition, various clinical nutritional therapies are used to treat Crohn's disease. Doctors treating inflammatory bowel disease can now offer several diets with more flexibility than ever. The Crohn's disease exclusion diet is a widely used diet for patients with active Crohn's disease. The Crohn's disease exclusion diet requires both exclusion and inclusion. Periodic exclusion of harmful foods and inclusion of wholesome foods gradually improves a patient's nutritional status. This article reviews the Crohn's disease exclusion diet, including its structure, mechanisms, research findings, and clinical applications.


The Efficacy of Medial Patellofemoral Ligament Reconstruction Combined with Tibial Tuberosity Transfer in the Treatment of Patellofemoral Instability.
A systematic review of the literature was undertaken to evaluate the efficacy of medial patellofemoral ligament (MPFL) reconstruction combined with tibial tuberosity transfer (TTT) in the treatment of patellofemoral instability. Using PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, a systematic search was carried out to identify and review the published literature pertinent to MFPL reconstruction combined with TTT. Relevant studies were critically appraised with narrative data synthesis. Studies that met the eligibility criteria were suitable for appraisal and consisted of case series and therapeutic series (levels IV & III). All studies had inherent variations in outcomes reporting and limited follow-up. Combined treatment offers restoration of normal anatomy, thus adding clinical value to the currently recommended anatomic approach to MPFL reconstruction. Nevertheless, the current body of evidence does not determine the threshold at which patellofemoral axis requires the need for adjunctive distal realignment as opposed to MPFL reconstruction alone. This review highlighted numerous recurring limitations in the conduct and presentation of the studies, which inadvertently mitigated the interpretation of their results. Future priority should be awarded to larger randomised controlled trials utilising validated patient reported outcome measures.


Synchrotron-produced ultrasoft X-rays: a tool for testing biophysical models of radiation action.
Ultrasoft X-rays are useful for mechanistic studies of ionizing radiation damage in living cells due to the localized nature of their energy depositions. To date radiobiology experiments in this energy region have relied on characteristic X-rays (mainly Alk and Ck) from X-ray tubes. However, limitations in the photon intensity and the available energies from X-ray tube sources prevent a definitive characterization of the relationship between photon energy and biological damage. Synchrotron radiation has the potential to avoid these limitations, since it produces X-rays with high intensity over a continuous spectrum. We have established a synchrotron-based system for radiation biology studies using the ES-0 exposure station of the Center for X-ray Lithography at the University of Wisconsin Synchrotron Radiation Center storage ring, Aladdin. A characterization of the system including spectral and intensity properties of the photon beam is presented. The first mammalian cell survival curve for synchrotron-produced ultrasoft X-rays was generated and is presented. Cell survival curves of C3H/10T 1/2 cells using synchrotron radiation of 1.48 keV agree with previous data using Alk X-rays (1.49 keV). An RBE of 1.47 +/- 0.30 at the 10% survival level was measured with reference to 250 kVp X-rays.


Single-molecule quantification of 5-hydroxymethylcytosine for diagnosis of blood and colon cancers.
The DNA modification 5-hydroxymethylcytosine (5hmC) is now referred to as the sixth base of DNA with evidence of tissue-specific patterns and correlation with gene regulation and expression. This epigenetic mark was recently reported as a potential biomarker for multiple types of cancer, but its application in the clinic is limited by the utility of recent 5hmC quantification assays. We use a recently developed, ultra-sensitive, fluorescence-based single-molecule method for global quantification of 5hmC in genomic DNA. The high sensitivity of the method gives access to precise quantification of extremely low 5hmC levels common in many cancers. We assessed 5hmC levels in DNA extracted from a set of colon and blood cancer samples and compared 5hmC levels with healthy controls, in a single-molecule approach. Using our method, we observed a significantly reduced level of 5hmC in blood and colon cancers and could distinguish between colon tumor and colon tissue adjacent to the tumor based on the global levels of this molecular biomarker. Single-molecule detection of 5hmC allows distinguishing between malignant and healthy tissue in clinically relevant and accessible tissue such as blood and colon. The presented method outperforms current commercially available quantification kits and may potentially be developed into a widely used, 5hmC quantification assay for research and clinical diagnostics. Furthermore, using this method, we confirm that 5hmC is a good molecular biomarker for diagnosing colon and various types of blood cancer.


PEGylated VRB plus quinacrine cationic liposomes for treating non-small cell lung cancer.
Non-small cell lung cancer (NSCLC) is the most common form of lung cancer, and the treatment effects are usually unsatisfactory. Vinorelbine (VRB) is extensively used in cancer treatment, but it has some disadvantages when used alone. PEGylated liposomes have been extensively used as a delivery carrier for antitumor drugs via prolonging the circulation time in the blood. The nanostructured liposomes were designed and prepared for treating NSCLC. In the liposomes, PEG was modified on the liposomal surface, DC-Chol was used as cationic materials, and VRB plus quinacrine were encapsulated in an aqueous core of the liposomes as an antitumor drug and an apoptosis-inducing agent, respectively. Evaluations were performed on A549 cells, tubular network formations and xenografts of the A549 cells. The PEGylated drugs-loaded cationic liposomes could significantly enhance cellular uptake and selectively accumulate in A549 cells, thus leading to show strongest antitumor efficacy to tumor cells and to tumor-bearing mice. Action mechanisms showed that the enhanced efficacy in treating NSCLC was related to activate caspase 9 and caspase 3, to activate Bax and P53, and to suppress Bcl-2 and Mcl-1. The PEGylated VRB plus quinacrine cationic liposomes showed a potential strategy for treating NSCLC.


A rapid capillary gas chromatographic method for the determination of propionic acid and propionates in bread and bread products.
A rapid method for the determination of propionic acid and propionates in bread and bread products is reported. The method employs solvent extraction, filtration and quantification by capillary gas chromatography using an internal standard. The limit of determination was 10 mg kg-1 propionic acid. The mean recovery rates for spiked bread samples were 108% at approximately 1,000 mg kg-1 (range of 106-110%, coefficient of variation 1.1%), 97% at approximately 2,000 mg kg-1 (range 96-98%, coefficient of variation 0.7%) and 98% at approximately 3,000 mg kg-1 (range of 97-100% and coefficient of variation of 1.1%). The method offers improved sensitivity, precision, chromatographic resolution and a wider sample analyte concentration range over a previously reported packed-column gas chromatographic method and has been applied to the analysis of bread, rolls, pitta breads and hamburger buns.


Trimodal speech perception: how residual acoustic hearing supplements cochlear-implant consonant recognition in the presence of visual cues.
As cochlear implant (CI) acceptance increases and candidacy criteria are expanded, these devices are increasingly recommended for individuals with less than profound hearing loss. As a result, many individuals who receive a CI also retain acoustic hearing, often in the low frequencies, in the nonimplanted ear (i.e., bimodal hearing) and in some cases in the implanted ear (i.e., hybrid hearing) which can enhance the performance achieved by the CI alone. However, guidelines for clinical decisions pertaining to cochlear implantation are largely based on expectations for postsurgical speech-reception performance with the CI alone in auditory-only conditions. A more comprehensive prediction of postimplant performance would include the expected effects of residual acoustic hearing and visual cues on speech understanding. An evaluation of auditory-visual performance might be particularly important because of the complementary interaction between the speech information relayed by visual cues and that contained in the low-frequency auditory signal. The goal of this study was to characterize the benefit provided by residual acoustic hearing to consonant identification under auditory-alone and auditory-visual conditions for CI users. Additional information regarding the expected role of residual hearing in overall communication performance by a CI listener could potentially lead to more informed decisions regarding cochlear implantation, particularly with respect to recommendations for or against bilateral implantation for an individual who is functioning bimodally. Eleven adults 23 to 75 years old with a unilateral CI and air-conduction thresholds in the nonimplanted ear equal to or better than 80 dB HL for at least one octave frequency between 250 and 1000 Hz participated in this study. Consonant identification was measured for conditions involving combinations of electric hearing (via the CI), acoustic hearing (via the nonimplanted ear), and speechreading (visual cues). The results suggest that the benefit to CI consonant-identification performance provided by the residual acoustic hearing is even greater when visual cues are also present. An analysis of consonant confusions suggests that this is because the voicing cues provided by the residual acoustic hearing are highly complementary with the mainly place-of-articulation cues provided by the visual stimulus. These findings highlight the need for a comprehensive prediction of trimodal (acoustic, electric, and visual) postimplant speech-reception performance to inform implantation decisions. The increased influence of residual acoustic hearing under auditory-visual conditions should be taken into account when considering surgical procedures or devices that are intended to preserve acoustic hearing in the implanted ear. This is particularly relevant when evaluating the candidacy of a current bimodal CI user for a second CI (i.e., bilateral implantation). Although recent developments in CI technology and surgical techniques have increased the likelihood of preserving residual acoustic hearing, preservation cannot be guaranteed in each individual case. Therefore, the potential gain to be derived from bilateral implantation needs to be weighed against the possible loss of the benefit provided by residual acoustic hearing.


European validation of the Vancouver classification of periprosthetic proximal femoral fractures.
The Vancouver classification has been shown by its developers to be a valid and reliable method for categorising the configuration of periprosthetic proximal femoral fractures and for planning their management. We have re-validated this classification system independently using the radiographs of 30 patients with periprosthetic fractures. These were reviewed by six experienced consultant orthopaedic surgeons, six trainee surgeons and six medical students in order to assess intra- and interobserver reliability and reproducibility. Each observer read the radiographs on two separate occasions. The results were subjected to weighted kappa statistical analysis. The respective kappa values for interobserver agreement were 0.72 and 0.74 for consultants, 0.68 and 0.70 for trainees on the first and second readings of the radiographs and 0.61 for medical students. The intra-observer agreement for the consultants was 0.64 and 0.67, for the trainees 0.61 and 0.64, and for the medical students 0.59 and 0.60 for the first and second readings, respectively. The validity of the classification was studied by comparing the pre-operative radiological findings within B subgroups with the operative findings. This revealed agreement for 77% of these type-B fractures, with a kappa value of 0.67. Our data confirm the reliability and reproducibility of this classification system in a European setting and for inexperienced staff. This is a reliable system which can be used by non-experts, between centres and across continents.


Growth of fetal thymocytes in organ culture: effect of recombinant lymphokines on thymocyte maturation.
Interleukin 2 (IL2) has a dose-dependent inhibitory effect on the growth and phenotypic maturation of thymocyte populations grown in fetal thymus organ culture. Addition of IL2 (100 U/ml) to 14-day fetal thymus organ cultures induces the appearance of a population of lymphokine-activated killer (LAK) cells which lyse allogeneic, syngeneic, and syngeneic tumor cell targets. The addition of the monoclonal antibody, PC-61, blocks the IL2-dependent growth and activation of LAK cells but does not influence the maturation of CD4+ CD8+ fetal thymocytes. These data imply that IL2 is not a major regulator of normal fetal thymocyte maturation. The effects of a range of recombinant lymphokines (IL1 alpha, IL1 beta, IL3, IL4, GM-CSF, G-CSF, M-CSF) on the proliferation and phenotypic maturation of fetal 14-day thymocytes in organ culture has been analysed. Two significant changes were seen. First, IL1 alpha and IL1 beta inhibited growth and the expression of the CD4 and CD8 antigens in organ culture, and second, GM-CSF increased the expression of Mac-1+ cells. IL4, which has known T cell growth-promoting activity, IL3, G-CSF, and M-CSF did not alter either normal growth or surface antigen expression in fetal thymocytes. While some of these lymphokines may function as accessory molecules in fetal thymocyte development, our experiments suggest that they do not have a significant influence on thymocyte maturation when used alone.


Chlorinated Polyfluorinated Ether Sulfonates Exhibit Higher Activity toward Peroxisome Proliferator-Activated Receptors Signaling Pathways than Perfluorooctanesulfonate.
Chlorinated polyfluorinated ether sulfonates (Cl-PFAESs) are the alternative products of perfluorooctanesulfonate (PFOS) in the metal plating industry in China. The similarity in chemical structures between Cl-PFAESs and PFOS makes it reasonable to assume they possess similar biological activities. In the present study, we investigated whether Cl-PFAESs could induce cellular effects through peroxisome proliferator-activated receptors (PPARs) signaling pathways like PFOS. By using fluorescence competitive binding assay, we found two dominant Cl-PFAESs (6:2 Cl-PFAES and 8:2 Cl-PFAES) bound to PPARs with affinity higher than PFOS. Based on the luciferase reporter gene transcription assay, the two Cl-PFAESs also showed agonistic activity toward PPARs signaling pathways with potency similar to (6:2 Cl-PFAES) or higher than (8:2 Cl-PFAES) PFOS. Molecular docking simulation showed the two Cl-PFAESs fitted into the ligand binding pockets of PPARs with very similar binding mode as PFOS. The cell function results showed Cl-PFAESs promoted the process of adipogenesis in 3T3-L1 cells with potency higher than PFOS. Taken together, we found for the first time that Cl-PFAESs have the ability to interfere with PPARs signaling pathways, and current exposure level of 6:2 Cl-PFAES in occupational workers has exceeded the margin of safety. Our study highlights the potential health risks of Cl-PFAESs as PFOS alternatives.


Family resemblance for anthropometric traits. II. Assessment of maternal occupational and age effects.
The present study was based on a cross-sectional sample of 1326 subjects (197 fathers, 466 mothers, 307 sons and 356 daughters) belonging to 488 nuclear families from the province of Biscay (Basque Country, Spain), with the purpose of estimating the degree of familial resemblance, for several anthropometric traits, by analysing the correlation coefficients between parent-offspring pairs. Height, weight, biacromial and bicrystal breadths, humerus and femur biepicondylar breadths, arm, waist and hip circumferences, biceps, triceps, subscapular, suprailiac, abdominal, thigh and calf skinfolds were taken from each individual. BMI, WHR and the sum of the seven skinfolds was computed. The mother's occupation and the age of offspring were taken into account, since the combination of all these factors might have an effect on familial resemblance. The mothers were classified into housewife (HM) and working mothers (WM). The offspring were divided into prepuberal, puberal and postpuberal subgroups. Standardised residuals were used to compute father-offspring (FO) and mother-offspring (MO) relations through correlation coefficients computed by maximum likelihood. The results confirm the influence of age on the correlations, since FO correlations revealed an increasing trend in HM's children for weight and another six variables as they grew older. On the other hand, the weight change tends to decrease with age in FO correlations within the WM group. Depending on mother's occupation and children's age, the global trend in the sample results in higher correlations in the second group (WM) than in the first one (HM) for the whole age range, but specially in FO correlations before puberty, where four variables (weight, bicrystal breadth, triceps and subscapular skinfolds) yield statistically significant differences.


Reducing wait time for cataract surgery: comparison of 2 historical cohorts of patients in Montreal.
A cataract efficiency program was implemented in Montreal in 2003 to decrease surgery wait time. Our goal was to determine whether health, adverse events during wait time, and outcome of patients presenting for cataract surgery differed from 1999 to 2006 in Montreal. Prospective preoperative and postoperative observational study performed at 2 time points 6 years apart. Patients awaiting first-eye cataract surgery at Maisonneuve-Rosemont Hospital: 509 patients in 1999-2000 and 206 patients in 2006-2007. Patients awaiting first-eye cataract surgery were recruited from Maisonneuve-Rosemont Hospital in 1999-2000 (n = 509) and a second cohort was recruited in 2006-2007 (n = 206). Date of entry onto the hospital waiting list and date of cataract surgery were recorded. About 2 weeks before surgery, patients were asked about accidents and falls while waiting, visual difficulty, and satisfaction with vision and wait time. Visual acuity was measured in each eye. Patients also completed interviewer-administered questionnaires: the 5-item Cataract Symptom Scale, Visual Function-14 Questionnaire (VF-14), Short Form Health Survey-36, Geriatric Depression Scale, and the 14-item Systemic Comorbidity Scale. The interview was repeated after surgery. In 1999, 39% of patients waited more than 6 months for cataract surgery, and this was reduced to 29% in 2006. Patients had better preoperative visual acuity in the surgical eye, less visual difficulty, and fewer cataract symptoms, and reported fewer accidents while waiting for surgery in 2006. The change in visual acuity after surgery was nonetheless the same in the 2 cohorts. The 2006 cohort achieved significantly higher VF-14 scores and reported more satisfaction with vision after surgery than did the 1999 cohort. Patients had cataract surgery sooner in the disease process in 2006-2007 compared with 1999-2000, with changes in visual acuity after surgery that were clinically significant in both cohorts.


Management of Groin Pain Using an Iliohypogastric Nerve Block in a Patient with Inguinal Hernia due to Persistent Müllerian Duct Syndrome.
Persistent Müllerian duct syndrome can cause an inguinal hernia, although this is a rare occurrence; recurrent inguinal hernias can, in turn, cause ongoing groin pain. Management of groin pain plays an important role in patients' quality of life. We present our experience with a 43-year-old man who had a 2-week history of left-sided groin pain. The patient underwent laparoscopic surgery for a left inguinal hernia via the transabdominal preperitoneal approach. Right-sided cryptorchidism was noted during surgery, with a solid structure-thought to be a uterus-extending into the left inguinal canal. The diagnosis was persistent Müllerian duct syndrome, and the groin pain was relieved after a laparoscopic right orchiectomy with a bilateral preperitoneal hernia repair using a mesh. Four years later, magnetic resonance imaging performed for new-onset left groin pain showed a left inguinal hernia caused by the uterine structure. We diagnosed the recurrent hernia as the cause of his pain. Prior to performing any invasive surgical procedures, an iliohypogastric nerve block was performed using 1% lidocaine. Short-term analgesia was provided by the block, improving his quality of life. He has been followed since then and has declined surgical neurectomy. An iliohypogastric nerve block can be an effective method of controlling groin pain caused by an inguinal hernia resulting from persistent Müllerian duct syndrome; the effectiveness of the nerve block will help determine whether surgical neurectomy is indicated for permanent pain control.


The effect of sage, sodium erythorbate and a mixture of sage and sodium erythorbate on the quality of turkey meatballs stored under vacuum and modified atmosphere conditions.
1. The combined effect of sage (S), sodium erythorbate (SE), a mixture of sage and sodium erythorbate (MIX) and vacuum packaging (VP) and modified atmosphere packaging (MAP) on the quality of cooked turkey meatballs stored at 4°C was investigated. The physicochemical properties (colour, MDA, AV, pH, water activity), microbiological quality characteristics (counts of mesophilic and psychrotrophic bacteria, fungi, coliforms and Clostridium sp.) and flavour attributes of meatballs were determined. 2. The values of the colour parameters L*, a* and b* were affected by the additives and packaging method. The colour of meatballs was better protected by sodium erythorbate than by sage or a mixture of sage and sodium erythorbate. The additives effectively stabilised lipids against oxidation and slowed down hydrolytic changes in turkey meatballs. Sage and a mixture of sage and sodium erythorbate showed stronger antioxidant properties than sodium erythorbate added alone. Products with additives were characterised by better sensory quality than control samples. Sage and MIX prevented the growth of mesophilic and psychrotrophic bacteria. All additives inhibited the growth of coliforms. 3. MAP was more effective than VP in maintaining the microbial and sensory quality stability of cooked turkey meatballs. However, VP appears to be a better method as regards the maintaining of lipid stability in turkey meatballs.


Bendable Photodetector on Fibers Wrapped with Flexible Ultrathin Single Crystalline Silicon Nanomembranes.
Silicon (Si) nanomembranes (NMs) enable conformal covering on complicated surfaces for novel applications. We adopt classical fibers as flexible/curved substrates and wrap them with freestanding ultrathin Si-NMs with a thickness of ∼20 nm. Intrinsic defects in single-crystalline Si-NMs provide a flow path for hydrofluoric acid (HF) to release the NM with a consecutive area of ∼0.25 cm2. Such Si-NMs with ultralow flexural rigidities are transferred onto a single-mode fiber (SMF) and functionalized into bendable photodetectors, which detects the leaked light when the fiber is bent. Our demonstration exemplifies optoelectronic applications in flexible photodetector for Si-NMs in a three-dimensional (3D) geometry.


White matter fiber tractography via anisotropic diffusion simulation in the human brain.
A novel approach to noninvasively tracing brain white matter fiber tracts is presented using diffusion tensor magnetic resonance imaging (DT-MRI). This technique is based on successive anisotropic diffusion simulations over the human brain, which are utilized to construct three dimensional diffusion fronts. The fiber pathways are determined by evaluating the distance and orientation from the fronts to their corresponding diffusion seeds. Synthetic and real DT-MRI data are employed to demonstrate the tracking scheme. It is shown that the synthetic tracts are accurately replicated, and several major white matter fiber pathways can be reproduced noninvasively, with the tract branching being allowed. Since simulating the diffusion process, which is truly a physical phenomenon reflecting the underlying architecture of cerebral tissues, makes full use of the diffusion tensor data, including both the magnitude and orientation information, the proposed approach is expected to enhance robustness and reliability in white matter fiber reconstruction.


Physiological and structural tradeoffs underlying the leaf economics spectrum.
The leaf economics spectrum (LES) represents a suite of intercorrelated leaf traits concerning construction costs per unit leaf area, nutrient concentrations, and rates of carbon fixation and tissue turnover. Although broad trade-offs among leaf structural and physiological traits have been demonstrated, we still do not have a comprehensive view of the fundamental constraints underlying the LES trade-offs. Here, we investigated physiological and structural mechanisms underpinning the LES by analysing a novel data compilation incorporating rarely considered traits such as the dry mass fraction in cell walls, nitrogen allocation, mesophyll CO2 diffusion and associated anatomical traits for hundreds of species covering major growth forms. The analysis demonstrates that cell wall constituents are major components of leaf dry mass (18-70%), especially in leaves with high leaf mass per unit area (LMA) and long lifespan. A greater fraction of leaf mass in cell walls is typically associated with a lower fraction of leaf nitrogen (N) invested in photosynthetic proteins; and lower within-leaf CO2 diffusion rates, as a result of thicker mesophyll cell walls. The costs associated with greater investments in cell walls underpin the LES: long leaf lifespans are achieved via higher LMA and in turn by higher cell wall mass fraction, but this inevitably reduces the efficiency of photosynthesis.


Comparative metabolism of tussilagone in rat and human liver microsomes using ultra-high-performance liquid chromatography coupled with high-resolution LTQ-Orbitrap mass spectrometry.
Tussilagone is a major component in Tussilago farfara that has been widely used as an anti-tussive herbal medicine for the treatment of bronchitis, cough and asthmatic disorders in the clinic. However, its metabolism has been poorly investigated. In order to clarify its in vitro metabolism, a comparative analysis of its metabolic profile in rat liver microsomes (RLMs) and human liver microsomes (HLMs) was carried out. Further, the cytochrome P450 isoforms (CYPs) involved in the metabolism were investigated. In this work, the biotransformation of tussilagone in RLMs and HLMs was compared using ultra-high-performance liquid chromatography coupled with high-resolution LTQ-Orbitrap mass spectrometry (UHPLC/HRMS) and the CYPs involved in the metabolism were further investigated by recombinant human CYP enzymes. Totally, nine metabolites of tussilagone were identified in RLMs and HLMs based on the proposed MS/MS fragmentation pathways of tussilagone and the accurate MS/MS spectra. Among them, one metabolite (M9) was detected in both RLMs and HLMs while the other eight metabolites were only detected in HLMs. Three hydroxylation metabolites (M6, M7 and M8) were detected in the assay with individual recombinant P450s incubation. M6 was detected in all CYPs except CYP2A6 while M7 and M8 were only observed in CYP3A4. The HR-ESI-MS/MS fragmentation behavior of tussilagone and its metabolic profile in RLMs and HLMs were investigated for the first time. The results demonstrated that the biotransformation of tussilagone involved hydrolysis of ester bonds at C-14 and hydroxylation in the side chains at C-12, C-5' or C-6'. Among the CYPs, CYP3A4 played an important role in the hydroxylation reaction of tussilagone in vitro. Furthermore, the results indicated a species-related difference in the metabolism of tussilagone between RLMs and HLMs. This work provided basic information for the metabolism of tussilagone in RLMs and HLMs, which would help to better understand the pharmacological activities of tussilagone.


Smoking and diabetic nephropathy.
Forty-seven patients with insulin-dependent diabetes (IDDM) and diabetic nephropathy and 47 controls with IDDM without diabetic nephropathy were interviewed about their previous and current smoking habits. The patients in the two groups were matched according to sex, age, age at onset, and duration of diabetes. All patients in the nephropathy group had proteinuria and decreased glomerular filtration. None in the control group had ever had proteinuria as tested by dip stick. The total amount of smoking until date of interview was estimated for each individual and presented as an index. The patients with nephropathy had a significantly higher smoking index than the controls. In the nephropathy group there were also more numerous current smokers, more heavy smokers and fewer individuals who had never smoked than in the control group. The link between diabetic renal microangiopathy and smoking may be through mechanisms such as increased platelet aggregation, accentuated tissue hypoxia and hemodynamic or metabolic effects of repeated noradrenaline release.


Efficient Noninvasive FHB Estimation using RGB Images from a Novel Multiyear, Multirater Dataset.
Fusarium head blight (FHB) is one of the most prevalent wheat diseases, causing substantial yield losses and health risks. Efficient phenotyping of FHB is crucial for accelerating resistance breeding, but currently used methods are time-consuming and expensive. The present article suggests a noninvasive classification model for FHB severity estimation using red-green-blue (RGB) images, without requiring extensive preprocessing. The model accepts images taken from consumer-grade, low-cost RGB cameras and classifies the FHB severity into 6 ordinal levels. In addition, we introduce a novel dataset consisting of around 3,000 images from 3 different years (2020, 2021, and 2022) and 2 FHB severity assessments per image from independent raters. We used a pretrained EfficientNet (size b0), redesigned as a regression model. The results demonstrate that the interrater reliability (Cohen's kappa, κ) is substantially lower than the achieved individual network-to-rater results, e.g., 0.68 and 0.76 for the data captured in 2020, respectively. The model shows a generalization effect when trained with data from multiple years and tested on data from an independent year. Thus, using the images from 2020 and 2021 for training and 2022 for testing, we improved the F1w score by 0.14, the accuracy by 0.11, κ by 0.12, and reduced the root mean squared error by 0.5 compared to the best network trained only on a single year's data. The proposed lightweight model and methods could be deployed on mobile devices to automatically and objectively assess FHB severity with images from low-cost RGB cameras. The source code and the dataset are available at https://github.com/cvims/FHB_classification.


Overexpression of clusterin correlates with tumor progression, metastasis in gastric cancer: a study on tissue microarrays.
Clusterin (CLU) is expressed in a wide variety of human tissues and fluids. Overexpression of cytoplasmic clusterin (sCLU) has been implicated in cancer development and progression. The aim of the present study is to evaluate the association of sCLU overexpression with clinicopathological features of human gastric carcinomas (GC).We constructed a gastric cancer tissue microarray containing 173 primary gastric carcinomas and 70 paired non-neoplastic mucosa specimens. The expression of sCLU was studied by immunohistochemistry. The correlations between sCLU expression and clinicopathological features, p53 abnormality, as well as Ki67 activation were analyzed. Overexpressions of sCLU was detected in 28.5% (n=165) of primary GCs by immunohistochemical staining, but not in non-neoplastic mucosa. Clinical association study found that overexpression of sCLU was significantly correlated with lymph-node metastasis (p < 0.001), tumor invasion (p < 0.001) and TNM stage (p < 0.001). In Kaplan-Meier survival analysis, overexpression of sCLU was significantly correlated with unfavorable survival in advanced GCs (p < 0.03). Furthermore, the association of sCLU with abnormal expression of p53 was ascertained. These results suggested that overexpression of sCLU was involved in the progression of GC and it's oncogenic function might be associated with p53 abnormality. Overexpression of sCLU seems to be related with patient's shorter survival in late stage GC.


Effects of alpha 1-adrenergic receptor subtype selective antagonists on lower urinary tract function in rats with bladder outlet obstruction.
Antagonists of alpha 1-adrenergic receptors (alpha 1ARs) relieve obstructive and irritative symptoms in patients with bladder outlet obstruction. However, to our knowledge mechanisms underlying the relief of irritative symptoms remain unknown. Because bladder alpha 1dARs are up-regulated in some rats with bladder outlet obstruction, we investigated the effect of the alpha 1aAR antagonist 5-methyl urapidil (5MU) vs the alpha 1a/alpha 1dAR antagonist tamsulosin on urinary frequency in obstructed rats. Baseline frequency was measured using a chronic micturition recording system and then obstruction (40 rats) or sham obstruction surgery (11 rats) was performed. After 6 weeks frequency was reassessed, followed by subcutaneous implantation of osmotic pumps to deliver 5MU, tamsulosin or vehicle for 1 week. Upon the completion of drug treatment urinary frequency was again measured and the pressor response to the alpha 1AR agonist phenylephrine was documented. Obstructed bladder mass was an average of 4.9 times greater than bladder mass in sham operated rats (p <0.001). Urinary frequency was elevated in obstructed rats with a bladder mass of greater than 500 mg vs all rats with a bladder mass of under 255 mg (p = 0.01). Of rats with a bladder mass of greater than 500 mg frequency was decreased in those treated with tamsulosin (p = 0.03) but not in those treated with 5MU. Tamsulosin and 5MU inhibited the pressor response to phenylephrine. Urinary frequency is increased in rats with a bladder mass of greater than 500 mg. The combined alpha 1a/alpha 1dAR antagonist tamsulosin decreases urinary frequency more than the alpha 1aAR selective antagonist 5MU. This finding supports the hypothesis that the alpha 1dAR is important for mediating irritative symptoms.


Improved reagent for trimethylsilylation of sphingolipid bases.
This paper describes the trimethylsilylation of sphingolipid bases under conditions that give derivatives of improved stability. The retention times of the common C(18) and C(20) long-chain bases, including the anhydro bases, obtained on a commercially available gas-liquid chromatographic column with a nonpolar stationary phase are given. Data are also presented on the separation of the erythro and threo isomers of sphingosine and dihydrosphingosine, as the trimethylsilyl ethers of the N-acetyl derivatives.


Melanoma in the elderly patient.
The incidence of cutaneous melanoma is rising steadily, and the rate of increase is among the highest for any form of cancer. Although the reliability of age as a prognostic factor is debatable, several studies suggest that age has an important prognostic use. Age alone does not predict a poor prognosis in the older patient with melanoma. University teaching hospital. A retrospective review was undertaken to identify patients aged 65 years or older with intermediate-thickness melanoma (1-4 mm). Two hundred thirteen such patients were identified. Data are given as mean +/- SD. The mean age was 72.2+/-6.1 years. The mean follow-up was 49 months. By univariate analysis, the mean disease-free survival (DFS) and overall survival (OS) for lymph node-positive patients was 36.0+/-9.6 and 56.0+/-10.6 months, respectively. The mean DFS for node-negative patients was 155.0+/-9.8 months, and the mean OS was 166.0+/-9.2 months (P<.001 for both). The mean DFS and OS for women were 151.0+/-11.2 and 163.0+/-10.9 months, respectively. In contrast, men had 116.0+/-9.5 months' DFS and 127.0+/-9.0 months' OS (P=.01 for both). By multivariate analysis, lymph node status was the most predictive variable for DFS and OS (P<.001 for both). Sex tended to affect OS (P=.02) but did not achieve prognostic significance on DFS (P=.09). Other factors such as location, ulceration, histological type, and mitoses per square millimeter failed to show any prognostic significance. Stratification into 3 age groups (65-70, 71-80, and >80 years) had no significant effect on DFS (P=.95) or OS (P=.92). Lymph node status is the most important prognostic factor in older patients with intermediate-thickness melanoma. Identification of high-risk factors may help stratify these patients for recommendation of more aggressive treatment or adjuvant therapies. Among these patients, age alone was not a significant prognostic factor in the clinical management of melanoma.


[Modern instrumentation for the studies of isoflavones].
Isoflavones belong to the natural substances exhibiting a number of physiological effects in living organisms. The substances are synthesized in plant tissues as protective agents against biotic stress (i.e. bacterial infection). Isoflavones are also an important dietary constituent in human nutrition. The review discusses modern trends in the studies of isoflavones in plant materials and foodstuffs and procedures for chemical analyses of isoflavones in human body fluids and plant tissues. Highly effective extraction and purification techniques, i.e. solid-phase extraction (SFE), accelerated-solvent extraction (ASE), and Soxhlet extraction, are presented. Latest procedures in chromatographic separation of isoflavones that apply different types of stationary phases are described. Immunochemical analysis, electrochemical sensing of isoflavones, spectrometric and other analytical techniques and their applications are also mentioned. Special attention is focused on a highly selective and sensitive technique of mass spectrometry and its application for identification of isoflavones and their glucosides in plants. Studies of interactions of isoflavones with cell receptors and a number of biologically active substances such as DNA and proteins are described. The reason for the presentation of the review was not to give a full overview of the presented topics but mainly to show modern and the most recent methods in the studies of isoflavones.


[Effects of SP1-11 and its N-terminal fragment SP1-4 on several indices of the microcirculatory system in stress].
In the experiments on rats it was shown that 5-hour immobilization induced the disturbances of terminal blood flow, degranulation of mast cells, increase of venular permeability and contractile activity of lymphaticus. I/p injection SP1-11 (125 micrograms/kg) before stress aggravated the disturbances caused by immobilization. The prophylactic i/p injection SP1-4 induced tranquilizing (100%), sedative (30%) or narcosis (20%) effect. In the rats with sedative or narcosis effects the relative normalization of components of microcirculatory system was observed.


Randomized clinical trial assessing the safety and immunogenicity of oral microencapsulated enterotoxigenic Escherichia coli surface antigen 6 with or without heat-labile enterotoxin with mutation R192G.
An oral, microencapsulated anti-colonization factor 6 antigen (meCS6) vaccine, with or without heat-labile enterotoxin with mutation R192G (LT(R192G)) (mucosal adjuvant), against enterotoxigenic Escherichia coli (ETEC) was evaluated for regimen and adjuvant effects on safety and immunogenicity. Sixty subjects were enrolled into a three-dose, 2-week interval or four-dose, 2-day interval regimen. Each regimen was randomized into two equal groups of meCS6 alone (1 mg) or meCS6 with adjuvant (2 microg of LT(R192G)). The vaccine was well tolerated and no serious adverse events were reported. Serologic response to CS6 was low in all regimens (0 to 27%). CS6-immunoglobulin A (IgA) antibody-secreting cell (ASC) responses ranged from 36 to 86%, with the highest level in the three-dose adjuvanted regimen; however, the magnitude was low. As expected, serologic and ASC LT responses were limited to adjuvanted regimens, with the exception of fecal IgA, which appeared to be nonspecific to LT administration. Further modifications to the delivery strategy and CS6 and adjuvant dose optimization will be needed before conducting further clinical trials with this epidemiologically important class of ETEC.


Nonadherence to beta blockers is a big issue with patients post-MI.
Although healthcare organizations have made great strides in boosting prescribing rates for beta blockers in patients who have suffered a myocardial infarction, (MI) data show that patients are not reaping the full value from these life-saving drugs. The problem is poor medication compliance. Experts maintain that providers and healthcare organizations should now turn their attention to this pressing problem.


Kocuria rhizophila prosthetic hip joint infection.
We present the first case report of prosthetic joint infection (PJI) caused by Kocuria rhizophila. Our patient is a 74-year-old male who underwent primary total hip replacement for right hip pain. His recovery was uneventful until 6 weeks postoperatively when he presented to his routine outpatient appointment with significant erythema, swelling, and tenderness over his right hip wound. Based on the acuity of his symptoms and the radiological findings, it was determined that the patient should undergo debridement, antibiotics, and implant retention (DAIR procedure). A consensus decision was also made at our PJI multidisciplinary meeting to treat him with 12 weeks of IV antibiotics. After completing this 12 weeks course of IV Vancomycin, his inflammatory markers returned to normal limits. At 6 months follow-up, our patient was mobilizing independently without any signs of infection recurrence. His radiographs showed the implant was in a satisfactory position with no evidence of loosening. This case adds to an emerging body of literature describing invasive infections associated with Kocuria species. We have demonstrated the effectiveness of managing this condition with debridement, implant retention, and IV Vancomycin therapy for 12 weeks.


Different growth factor requirements for human Th2 cells may reflect in vivo induced anergy.
We previously reported the isolation of allergen-specific Th2 lines and clones from atopy patch test (APT) sites of atopic dermatitis (AD) patients. Upon stimulation with allergen or anti-CD3+ phorbol myristate acetate (PMA) IL-4 was released with or without IL-5, while no (or extremely low concentrations of) IL-2 and interferon-gamma (IFN-gamma) were detectable. A high IL-4/IFN-gamma ratio facilitates production of allergen-specific IgE, of which high levels are observed in AD patients. Here we show that the above mentioned Th2 cells are notably different from murine Th2 cells. Not IL-4, which is the autocrine acting growth factor for murine Th2 cells, but IL-2 was needed for proliferation of these human APT-derived Th2 lines and clones. Of significance, unless exogenous IL-2 was added, no proliferative response to allergen, presented by Epstein-Barr virus-transformed B (EBV-B) cells, non-T cells or IgE-bearing Langerhans cells (LC), occurred. Lack of proliferation and IL-2 production after full T cell receptor (TCR) triggering is a characteristic first described for in vitro anergized T cells. However, like the clones we describe in this study, anergic T cells may retain production of cytokines other than IL-2. A further resemblance between anergic T cells and the human Th2 clones reported here is that IL-4 can enhance IL-2-driven proliferation, but is not capable of inducing T cell growth by itself. The absence of IL-4-driven proliferation differentiates human Th2 cells from murine Th2 cells. Both produce IL-4 when stimulated in a cognate fashion, but only murine Th2 cells will proliferate. We conclude that the presently reported human Th2 cells are different from murine Th2 cells, in that they need other T cells to produce IL-2 required for their expansion. Moreover, the Th2 cells phenotypically resemble anergic T cells. As yet, however, we have no clue as to whether these features account for the current Th2 cells only or for human Th2 cells in general. We hypothesize that the Th2 phenotype of AD skin-derived, allergen-specific T cells may be induced in vivo by LC, which lack CD80, and therefore do not provide secondary signals through CD28-CD80 interaction.


Membrane-mobility agent-promoted fusion of erythrocytes: fusibility is correlated with attack by calcium-activated cytoplasmic proteases on membrane proteins.
Rat, but not human, erythrocytes undergo fusion promoted by the membrane-mobility agent 2-(2-methoxyethoxy)-ethyl cis-8-(2-octylcyclopropyl)octanoate (A2C). The difference in behavior is correlated with rat erythrocyte membrane protein degradation caused by Ca2+-activated proteases. The human erythrocyte is deficient in such protease activity. Membrane protein degradation is a necessary, but not sufficient, requirement for membrane fusion. Membrane protein degradation probably releases membrane components from certain constraints. In addition, the motion of membrane components precedes fusion and must be promoted by reagents such as A2C, leading to the creation of fusion-potent lipid areas. This sequence of chemical and physical events occurs in other fusion processes.


Use of the Putamen as a Surrogate Anatomical Marker for the Internal Segment of the Globus Pallidus in Deep Brain Stimulation Surgery.
The success of deep brain stimulation (DBS) of the internal segment of the globus pallidus (GPi) depends on accurately placing the electrode into the GPi motor territory. Direct targeting can be difficult as GPi laminar borders are not always clearly identifiable on MRI. Here, we report a method for using the putamen (PUT) as a surrogate anatomical marker to target the GPi. We developed a PUT-based GPi targeting using the FGATIR (fast gray matter acquisition T1-weighted inversion recovery) MRI sequence and compared it with consensus coordinate-based indirect targeting. Stereotactic target coordinates were obtained and analyzed. In our GPi DBS case sequences, GPi borders were unresolvable on T2-weighted MRI. However, in all cases, application of the PUT-based method resulted in consistently localized GPi targets, which were confirmed by merging the T2-weighted MRI with the FGATIR MRI. Significant differences were noted in the target coordinates between the PUT-based method and indirect targeting based on both the distance from the anterior commissure and the distance from the intercommissural plane. The mean differences for mediolateral distance and anteroposterior distance were 1.4 and 1.42 mm, respectively. In addition, the PUT-based method estimated a target that was closer to the nearest implanted electrode. Our PUT-based method allows consistent and precise patient-specific GPi targeting. Further study is planned to correlate PUT-based GPi targeting with microelectrode recording, location of active contact of the DBS electrode and clinical outcome.


Stress Generation Research in Depression: A Commentary.
The concept of stress generation is a powerful tool that is consistent with existing cognitive-behavioral theories of depression. In this brief commentary on the literature on stress generation in depression, we highlight several issues that we believe will help to advance the stress generation field. Specifically, we discuss important methodological considerations, issues related to generality and specificity, and theoretical and clinical implications of stress generation. We then address common misperceptions of the stress generation hypothesis. Finally, we end by posing several questions about the mechanisms driving stress generation that should be addressed in future research. Advancing the field's knowledge about stress generation will yield a tangible direction for theory-driven, targeted intervention. Our hope is that this commentary will help to stimulate and frame future research in this exciting area.


Questions raised by a reasoned action approach: comment on Ogden (2003).
In her critique of social cognition or reasoned action models, J. Ogden (see record 2003-05896-016) claimed that such models are not falsifiable and thus cannot be tested, that the postulated relations among model components are true by definition, and that questionnaires used to test the models may create rather than assess cognitions and thus influence later behavior. The authors of this comment challenge all 3 arguments and contend that the findings Ogden regarded as requiring rejection of the models are, in fact, consistent with them, that there is good evidence for the validity of measures used to assess the models' major constructs, and that the effect of completing a questionnaire on cognitions and subsequent behavior is an empirical question.


Band Nesting Bypass in WS2 Monolayers via Förster Resonance Energy Transfer.
Two-dimensional (2D) transition-metal dichalcogenides (TMDs) have attracted intensive interest due to the direct-band-gap transition in the monolayer form, positioning them as potential next-generation materials for optoelectronic or photonic devices. However, the band-nested suppression of the recombination efficiency at higher excitation energies limits the ability to locally control and manipulate the photoluminescence of WS2 for multifunctional applications. In this work, we exploit an energy transfer method to modulate the fluorescence properties of TMDs under a larger excitation range spanning from UV to visible light. Self-assembled lanthanide (Ln)/TMD hybrids have been designed based on a low-cost and highly efficient solution-processed approach. The emission energy from Ln3+ sources can be effectively transferred to the TMD monolayers under low power exposure (0.13 mW) at room temperature, activating the characteristic monolayer fluorescence in place of Ln3+ emission signatures. The Ln/TMDs photonics can potentially tune the excitation of TMDs to provide variable yet controllable emissions. This provides a solution to the suppression of direct exciton recombination in monolayer TMDs at the band nesting resonant energy region. Our work on such Ln/TMD systems would overcome the limited excitation energy range in TMDs and extend their functionalities for optoelectronic or photonic applications.


[Stratification of coronary risk].
Coronary atheroselerosis constitutes the fifth leading cause of global disease burden and the leading cause in developed societies. Coronary artery disease is the leading cause of morbidity and mortality in the, United States, Western industrialized societies; in our country, it has become a real health problem. Compelling epidemiologic and clinical data have demonstrated that elevated total cholesterol levels, high blood pressure, and diabetes mellitus correlate with increasing coronary artery disease mortality. Risk factors modification in the general public and persons at high risk offers the best opportunity for effectively reducing the prevalence of coronary heart disease.


Morphological evolution of Cu2O nanocrystals in an acid solution: stability of different crystal planes.
The morphological evolution of uniform Cu(2)O nanocrystals with different morphologies in a weak acetic acid solution (pH = 3.5) has been studied for cubic, octahedral, rhombic dodecahedral, {100} truncated octahedral, and {110} truncated octahedral nanocrystals. Cu(2)O nanocrystals undergo oxidative dissolution in weak acid solution, but their morphological changes depend on the exposed crystal planes. We found that the stability of Cu(2)O crystal planes in weak acid solution follows the order of {100} ≫ {111} > {110} and determines how the morphology of Cu(2)O nanocrystals evolves. The stable {100} crystal planes remain, and new {100} facets form at the expense of the less stable {111} and {110} crystal planes on the surface of Cu(2)O nanocrystals. Density functional theory calculations reveal that the Cu-O bond on Cu(2)O(100) surface has the shortest bond length. These results clearly exemplify that the morphology of inorganic crystals will evolve with the change of local chemical environment, shedding light on fundamentally understanding the morphological evolution of natural minerals and providing novel insights into the geomimetic synthesis of inorganic materials in the laboratory.


Current recommendations for the diagnosis and treatment of concussion in sport: a comparison of three new guidelines.
Currently, there is considerable debate within the sports medicine community about the role of concussion and the risk of chronic neurological sequelae. This concern has led to significant confusion among primary care providers and athletic trainers about how to best identify those athletes at risk and how to treat those with concussion. During the first quarter of 2013, several new or updated clinical practice guidelines and position statements were published on the diagnosis, treatment, and management of mild traumatic brain injury/concussion in sports. Three of these guidelines were produced by the American Medical Society for Sports Medicine, The American Academy of Neurology, and the Zurich Consensus working group. The goal of each group was to clearly define current best practices for the definition, diagnosis, and acute and post-acute management of sports-related concussion, including specific recommendations for return to play. In this article, we compare the recommendations of each of the three groups, and highlight those topics for which there is consensus regarding the definition of concussion, diagnosis, and acute care of athletes suspected of having a concussion, as well as return-to-play recommendations.


Formation and growth of lipofuscin in the retinal pigment epithelium cells.
The kinetics of lipofuscin growth in diseased retinal pigment epithelium cells is investigated using Monte Carlo simulations and scaling theory on a cluster aggregation model. The model captures the essential physics of lipofuscin growth in the cells. A remarkable feature is that small particles may be removed from the cells while the larger ones become fixed and grow by aggregation. Model simulations are compared to the number of lipofuscin granules in eyes with early age-related degeneration.


[Sacral neuromodulation in patients with nonobstructive, chronic urinary retention: relevance of the carbachol test and influence of associated nerve lession].
Aim of this study was to evaluate whether the carbachol test and the nerve lesions responsible for bladder dysfunction possess predictive value in patients with chronic urinary retention who undergo temporary sacral neuromodulation (PNE test). In 1999 and 2000, PNE tests were performed in 24 patients with chronic urinary retention and acontractile detrusor previously assessed by urodynamics. In 18 patients, a carbachol test was performed during urodynamics. The diagnosis related to the acontractile detrusor was additionally assessed and compared to the successful outcome of the PNE test. The PNE test was successful in 8 of the 24 patients (33.3 %). The bladder was completely emptied during the PNE test in 3 of the 10 patients with negative carbachol test and in 3 of the 8 patients with positive carbachol test. The highest success rate (80 %) was observed in patients after hysterectomy, whereas after lumbosacral pulposal prolapse or with a CNS tumor, it was only 20 - 33.3 %. We conclude that sacral neuromodulation is an effective treatment option in patients with nonobstructive urinary retention. The carbachol test does not possess any definitive predictive value with respect to the success rate of sacral neuromodulation in patients with chronic urinary retention. The success rate more likely depends on the localisation of the nerve lesion. PNE tests should be performed in all patients with therapy resistent nonobstructive urinary retention, as no other predictive factors exist.


Probing of Actinobacillus pleuropneumoniae ApxIIIA toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells.
Actinobacillus pleuropneumoniae, the causative bacterial agent of porcine pleuropneumonia, produces Apx toxins which belong to RTX toxin family and are recognized as the major virulence factors. So far, their target receptor(s) has not been identified and the disease cytopathogenesis remains poorly understood. Production of an active Apx toxin and characterization of its toxic activity constitute the premises necessary to the description of its interaction with a potential receptor. From this point of view, we produced an active recombinant ApxIIIA toxin in order to characterize its toxicity on peripheral blood mononucleated cells (PBMCs) isolated from several species. Toxin preparation exercises a strong cytotoxic action on porcine PBMCs which is directly related to recombinant ApxIIIA since preincubation with polymyxin B does not modify the cytotoxicity rate while preincubation with a monospecific polyclonal antiserum directed against ApxIIIA does. The cell death process triggered by ApxIIIA is extremely fast, the maximum rate of toxicity being already reached after 20 minutes of incubation. Moreover, ApxIIIA cytotoxicity is species-specific because llama, human, dog, rat and mouse PBMCs are resistant. Interestingly, bovine and caprine PBMCs are slightly sensitive to ApxIIIA toxin too. Finally, ApxIIIA cytotoxicity is cell type-specific as porcine epithelial cells are resistant. We have produced an active recombinant ApxIIIA toxin and characterized its specific cytotoxicity on porcine PBMCs which will allow us to get new insights on porcine pleuropneumonia pathogenesis in the future.


Heterogeneous anomalous transport in cellular and molecular biology.
It is well established that a wide variety of phenomena in cellular and molecular biology involve anomalous transport e.g. the statistics for the motility of cells and molecules are fractional and do not conform to the archetypes of simple diffusion or ballistic transport. Recent research demonstrates that anomalous transport is in many cases heterogeneous in both time and space. Thus single anomalous exponents and single generalised diffusion coefficients are unable to satisfactorily describe many crucial phenomena in cellular and molecular biology. We consider advances in the field ofheterogeneous anomalous transport(HAT) highlighting: experimental techniques (single molecule methods, microscopy, image analysis, fluorescence correlation spectroscopy, inelastic neutron scattering, and nuclear magnetic resonance), theoretical tools for data analysis (robust statistical methods such as first passage probabilities, survival analysis, different varieties of mean square displacements, etc), analytic theory and generative theoretical models based on simulations. Special emphasis is made on high throughput analysis techniques based on machine learning and neural networks. Furthermore, we consider anomalous transport in the context of microrheology and the heterogeneous viscoelasticity of complex fluids. HAT in the wavefronts of reaction-diffusion systems is also considered since it plays an important role in morphogenesis and signalling. In addition, we present specific examples from cellular biology including embryonic cells, leucocytes, cancer cells, bacterial cells, bacterial biofilms, and eukaryotic microorganisms. Case studies from molecular biology include DNA, membranes, endosomal transport, endoplasmic reticula, mucins, globular proteins, and amyloids.


Influence of Nrf2 activators on subcellular skeletal muscle protein and DNA synthesis rates after 6 weeks of milk protein feeding in older adults.
In older adults, chronic oxidative and inflammatory stresses are associated with an impaired increase in skeletal muscle protein synthesis after acute anabolic stimuli. Conjugated linoleic acid (CLA) and Protandim have been shown to activate nuclear factor erythroid-derived 2-like 2 (Nrf2), a transcription factor for the antioxidant response element and anti-inflammatory pathways. This study tested the hypothesis that compared to a placebo control (CON), CLA and Protandim would increase skeletal muscle subcellular protein (myofibrillar, mitochondrial, cytoplasmic) and DNA synthesis in older adults after 6 weeks of milk protein feeding. CLA decreased oxidative stress and skeletal muscle oxidative damage with a trend to increase messenger RNA (mRNA) expression of a Nrf2 target, NAD(P)H dehydrogenase quinone 1 (NQO1). However, CLA did not influence other Nrf2 targets (heme oxygenase-1 (HO-1), glutathione peroxidase 1 (Gpx1)) or protein or DNA synthesis. Conversely, Protandim increased HO-1 protein content but not the mRNA expression of downstream Nrf2 targets, oxidative stress, or skeletal muscle oxidative damage. Rates of myofibrillar protein synthesis were maintained despite lower mitochondrial and cytoplasmic protein syntheses after Protandim versus CON. Similarly, DNA synthesis was non-significantly lower after Protandim compared to CON. After Protandim, the ratio of protein to DNA synthesis tended to be greater in the myofibrillar fraction and maintained in the mitochondrial and cytoplasmic fractions, emphasizing the importance of measuring both protein and DNA synthesis to gain insight into proteostasis. Overall, these data suggest that Protandim may enhance proteostatic mechanisms of skeletal muscle contractile proteins after 6 weeks of milk protein feeding in older adults.


Immunotherapy with interleukin-10 depends on the CXC chemokines inducible protein-10 and monokine induced by IFN-gamma.
The cytokine interleukin (IL)-10 has potent antitumor activity in many model systems when expressed locally at very high levels from the time of tumor transplantation. We now demonstrate that systemic administration of recombinant human IL-10 to animals bearing established highly malignant mammary tumors also leads to significant growth inhibition. We had shown previously that expression of the CXC chemokines Mig (monokine induced by IFN-gamma) and IP-10 (inducible protein 10) is observed in IL-10 transduced but not neo-vector control tumors. We now demonstrate that treatment of IL-10-tumor-bearing mice with antibodies to either chemokine partially reverses the therapeutic effect of IL-10. Tumor growth in animals treated with both antibodies is comparable with that of vector control tumors. Direct transduction of Mig cDNA into the parental tumor cell line before transplantation also results in smaller tumors. This tumor growth inhibition is associated with increased numbers of CD4+ cells, consistent with a T-cell chemoattractant activity for Mig. No change in vascularization, as indicated by CD31+ cells, was observed in either Mig or IL-10-transfected tumors. Thus, an antiangiogenic activity for either cytokine could not be confirmed. Mig and IP-10 are critical to the therapeutic response resulting from high levels of IL-10, and, furthermore, Mig as a single agent also has tumor-inhibitory activity in a model of breast cancer.


Parameters associated with cloning in Actinobacillus actinomycetemcomitans.
Characterization of virulence traits in Actinobacillus actinomycetemcomitans requires the application of recombinant DNA techniques. To develop appropriate genetic tools it is necessary to identify suitable host-vector systems. The current study assessed cloning parameters in A. actinomycetemcomitans for two previously described vectors, pDMG4 and pMMB67. It was determined that the maximum size of recombinant molecules that could be transferred to A. actinomycetemcomitans strain ATCC29522 via electroporation was 33 kb. The size limit for transformation of the same strain with ligation mixtures (direct cloning), however, was limited to 23-24 kb. Additional experiments included electroporation of various A. actinomycetemcomitans strains with plasmid DNA isolated from Escherichia coli and different A. actinomycetemcomitans sources. Differences in transformation efficiencies suggested the presence of a restriction modification system for pDMG4 in some strains of A. actinomycetemcomitans. Cloning of portions of the enterococcal plasmid pJH1 into A. actinomycetemcomitans resulted in the insertion of the intact vector into the chromosome.


Role of Anterior Intralaminar Nuclei of Thalamus Projections to Dorsomedial Striatum in Incubation of Methamphetamine Craving.
Relapse to methamphetamine (Meth) seeking progressively increases after withdrawal from drug self-administration (incubation of Meth craving). We previously demonstrated a role of dorsomedial striatum (DMS) dopamine D1 receptors (D1Rs) in this incubation. Here, we studied the role of afferent glutamatergic projections into the DMS and local D1R-glutamate interaction in this incubation in male rats. We first measured projection-specific activation on day 30 relapse test by using cholera toxin b (retrograde tracer) + Fos (activity marker) double-labeling in projection areas. Next, we determined the effect of pharmacological reversible inactivation of lateral or medial anterior intralaminar nuclei of thalamus (AIT-L or AIT-M) on incubated Meth seeking on withdrawal day 30. We then used an anatomical asymmetrical disconnection procedure to determine whether an interaction between AIT-L→DMS glutamatergic projections and postsynaptic DMS D1Rs contributes to incubated Meth seeking. We also determined the effect of unilateral inactivation of AIT-L and D1R blockade of DMS on incubated Meth seeking, and the effect of contralateral disconnection of AIT-L→DMS projections on nonincubated Meth seeking on withdrawal day 1. Incubated Meth seeking was associated with selective activation of AIT→DMS projections; other glutamatergic projections to DMS were not activated. AIT-L (but not AIT-M) inactivation or anatomical disconnection of AIT-L→DMS projections decreased incubated Meth seeking. Unilateral inactivation of AIT-L or D1R blockade of the DMS had no effect on incubated Meth craving, and contralateral disconnection of AIT-L→DMS projections had no effect on nonincubated Meth seeking. Our results identify a novel role of AIT-L and AIT-L→DMS glutamatergic projections in incubation of drug craving and drug seeking.SIGNIFICANCE STATEMENT Methamphetamine seeking progressively increases after withdrawal from drug self-administration, a phenomenon termed incubation of methamphetamine craving. We previously found that D1R-mediated dopamine transmission in the dorsomedial striatum plays a critical role in this incubation phenomenon. Here, we used neuroanatomical and neuropharmacological methods in rats to demonstrate that an interaction between the glutamatergic projection from the lateral anterior intralaminar nuclei of the thalamus to the dorsomedial striatum and local dopamine D1 receptors plays a critical role in relapse to methamphetamine seeking after prolonged withdrawal. Our study identified a novel motivation-related thalamostriatal projection critical to relapse to drug seeking.


A novel point-of-care diagnostic prototype system for the simultaneous electrochemiluminescent sensing of multiple traumatic brain injury biomarkers.
Traumatic brain injuries (TBI) are typically acquired when a sudden violent event causes damage to the brain tissue. A high percentage (70-85%) of all TBI patients are suffering from mild TBI (mTBI), which is often difficult to detect and diagnose with standard imaging tools (MRI, CT scan) due to the absence of significant lesions and specific symptoms. Recent studies suggest that a screening test based on the measurement of a protein biomarker panel directly from a patient's blood can facilitate mTBI diagnosis. Herein, we report a novel prototype system designed as a precursor of a future hand-held point-of-care (POC) diagnostic device for the simultaneous multi-biomarker sensing, employing a microarray-type spatially resolved electrochemiluminescence immunoassay (SR-ECLIA). The small tabletop prototype consists of a screen-printed electrode compartment to conduct multi-analyte ECL sandwich assays, a potentiostat module and a light collection module, all integrated into a compact 3D-printed housing (18.2 × 16.5 × 5.0 cm), as well as an sCMOS detector. Based on this design concept, further miniaturization, system integration, performance optimization and clinical evaluation shall pave the way towards the development of a portable instrument for use at the site of accident and healthcare. To demonstrate the system's feasibility, current performance and efficiency, the simultaneous detection of three mTBI biomarkers (GFAP, h-FABP, S100β) in 50% serum was achieved in the upper pg mL-1 range. The proposed device is amenable to the detection of other biomarker panels and thus could open new medical diagnostic avenues for sensitive multi-analyte measurements with low-volume biological sample requirements.


Effectiveness of intravenous ferric carboxymaltose in improving hemoglobin level among postpartum women with moderate-to-severe anemia at a secondary care hospital in Faridabad, Haryana - An interventional study.
Postpartum anemia is associated with postpartum anxiety, depression, poor cognitive interaction, and delayed infant development. Oral iron is the currently recommended therapy but is associated with gastrointestinal complaints and poor compliance. Parenteral iron dextran may cause serious side effects, whereas iron sucrose requires repeated visits. Intravenous ferric carboxymaltose (FCM) is given as an infusion in a single setting, which is logistically convenient both to patients and health system. It can be particularly beneficial for postpartum women who are usually discharged from health facility 48 h after delivery. The objectives of this study were to estimate the change in mean hemoglobin (Hb) level 6 weeks after administration of intravenous FCM and to document any adverse events following administration among postpartum women. A prospective, interventional follow-up study was conducted from August to December 2018. One hundred eligible postpartum women with Hb level of 5-9.9 g/dl were administered a calculated dose of intravenous FCM based on Ganzoni's formula with Hb target of 12 g/dl. Hb and serum ferritin were measured at baseline and at 6 weeks after FCM infusion and compared for mean increase. There was a significant increase in Hb and serum ferritin levels after administration of FCM. The mean (95% confidence interval) increase in the Hb and serum ferritin after 6 weeks was 4.2 (3.9-4.5) g/dl and 137.3 (113.6-161.0) ng/ml, respectively. No major adverse events were reported. Administration of FCM was safe and effective in the treatment of moderate-to-severe anemia among postpartum women.


The Clinical Features, Risk Factors, and Surgical Treatment of Cervicogenic Headache in Patients With Cervical Spine Disorders Requiring Surgery.
To clarify the clinical features and risk factors of cervicogenic headache (CEH; as diagnosed according to the International Classification of Headache Disorders-Third Edition beta) in patients with cervical spine disorders requiring surgery. CEH is caused by cervical spine disorders. The pathogenic mechanism of CEH is hypothesized to involve a convergence of the upper cervical afferents from the C1, C2, and C3 spinal nerves and the trigeminal afferents in the trigeminocervical nucleus of the upper cervical cord. According to this hypothesis, functional convergence of the upper cervical and trigeminal sensory pathways allows the bidirectional (afferent and efferent) referral of pain to the occipital, frontal, temporal, and/or orbital regions. Previous prospective studies have reported an 86-88% prevalence of headache in patients with cervical myelopathy or radiculopathy requiring anterior cervical surgery; however, these studies did not diagnose headache according to the International Classification of Headache Disorders criteria. Therefore, a better understanding of the prevalence rate, clinical features, risk factors, and treatment responsiveness of CEH in patients with cervical spine disorders requiring surgery is necessary. We performed a single hospital-based prospective cross-sectional study and enrolled 70 consecutive patients with cervical spine disorders such as cervical spondylotic myelopathy, ossification of the posterior longitudinal ligament, cervical spondylotic radiculopathy, and cervical spondylotic myeloradiculopathy who had been scheduled to undergo anterior cervical fusion or dorsal cervical laminoplasty between June 2014 and December 2015. Headache was diagnosed preoperatively according to the International Classification of Headache Disorders-Third Edition beta. The Japanese Orthopaedic Association Cervical Myelopathy Evaluation Questionnaire, Neck Disability Index, and a 0-100 mm visual analog scale (VAS) were used to evaluate clinical features, and scores were compared between baseline (ie, preoperatively) and 3, 6, and 12 months post-surgery. The prevalence of CEH in our population was 15/70 (21.4%, 95%CI: 11.8% to 31.0%). The main clinical features were dull and tightening/pressing headache sensations in the occipital region. Headache severity was mild (VAS, 32 ± 11 mm) and only one patient reported use of an oral analgesic. Compared to patients without CEH, patients with CEH had higher frequencies of neck pain (86.7% vs. 50.9%; P = .017), cervical range of motion limitation (ROM) (66.7% vs. 38.2%; P = .049), and higher Neck Disability Index scores (14 vs. 3; P < .001). Among the different cervical spine disorders, the prevalence of CEH was highest in cervical spondylotic myeloradiculopathy patients (60%), being ≤ 20% for all other disorders. Surgical treatments including cervical laminoplasty to relieve abnormal pressure on the spinal cord via a posterior approach, were associated with initial improvements in headache VAS that slightly diminished by 12 months post-surgery (P < .001). We report a lower prevalence of CEH in patients with cervical spinal disorders requiring surgery than that reported previously. The main clinical features of CEH were mild, dull, and tightening/pressing headache sensations in the occipital region. Potential risk factors for CEH included neck pain, limited cervical ROM, high Neck Disability Index score, and a diagnosis of cervical spondylotic myeloradiculopathy. The further accumulation of patients in a multi-institutional study may be required in order to discuss the diagnostic criteria and pathophysiology of this condition.


Global Pinning Synchronization of Complex Networks With Sampled-Data Communications.
This paper investigates the global pinning synchronization problem for a class of complex networks with aperiodic samplings. Combined with the Writinger-based integral inequality, a new less conservative criterion is presented to guarantee the global pinning synchronization of the complex network. Furthermore, a novel condition is proposed under which the complex network is globally pinning synchronized with a given performance index. It is shown that the performance index has a positive correlation with the upper bound of the sampling intervals. Finally, the validity and the advantage of the theoretic results obtained are verified by means of the applications in Chua's circuit and pendulum.


Atypical Ulnar Fracture in an Older Woman with Osteoporosis with a Five-Year History of Denosumab Treatment: A Case Report.
Atypical ulnar fracture (AUF) is a rare condition that may occur in patients treated with bisphosphonates. We describe the clinical course of a patient who had an AUF during treatment with denosumab. An 86-year-old woman treated with denosumab for 5 years experienced a complete transverse fracture of the proximal left ulna after falling from a standing height. Intraoperative photographs showed cortical thickening around the fracture site. Therefore, we diagnosed the patient with AUF and performed internal fixation with a locking plate. However, a re-fracture occurred 2.5 months after surgery, which was treated with locking plate replacement with a longer one with a bone graft. One year after the second surgery, the fracture healed. To our knowledge, this is the second case report of an AUF occurring during denosumab treatment. Our case highlights the importance of careful follow-up of patients receiving not only bisphosphonate, but also denosumab. Levels of Evidence: Level V (Therapeutic).


Efficacy and Safety of Anti-calcitonin Gene-Related Peptide (CGRP) Monoclonal Antibodies in Preventing Migraines: A Systematic Review.
The neuropeptide calcitonin gene-related peptide (CGRP) is an essential pathophysiological treatment for migraines. A unique class of medications called CGRP monoclonal antibodies target CGRP and its receptor and have demonstrated promising benefits in the treatment and prevention of migraines. This study sought to identify and assess the quality of existing systematic reviews about the effectiveness of CGRP antibodies for preventing migraines, as well as systematically review and synthesize the evidence on these topics. This included the four Food and Drug Administration (FDA)-approved medications erenumab, galcanezumab, fremanezumab, and eptinezumab. The effectiveness and safety of these monoclonal antibodies in preventing migraines should also be examined in light of patient characteristics, and any gaps in the body of knowledge should be noted in order to suggest new lines of investigation. Data gathering included a thorough search of internet databases (PubMed, Cochrane Library, Web of Science, and Scopus) for relevant research released between 2018 and 2023. The findings imply that CGRP monoclonal antibodies are efficient and secure for preventing migraines and may be considered a first-line alternative for treating migraines and drug misuse. The results further imply that combination treatment with CGRP antibodies and onabotulinumtoxinA may enhance the prevention of migraine in adults. With suggestions for more studies to find and address these variables, the significance of genetic and epigenetic factors in the progression of pediatric patients' acute postoperative pain to chronic postsurgical pain is underlined. All four anti-CGRP monoclonal antibodies, erenumab, fremanezumab, galcanezumab, and eptinezumab, were shown to be safe and effective for the prevention of migraine when the research additionally looked at their individual effectiveness and safety. Additionally, the study discovered considerable variances in effectiveness amongst various groups. However, further investigation is required to establish the best time and dosage and the effect of patient characteristics on the effectiveness and safety of these medications.


Synthesis of vinyl germylenes.
A disilylated germylene phosphine adduct was obtained by reaction of tris(trimethylsilyl)silyl potassium with PMe3 and GeCl2·dioxane. The adduct reacted with tolane to give a germacyclopropene, while reactions with phenylacetylene occurred by insertion into the Si-Ge bonds giving vinylgermylene adducts. Release of the mono- or divinylgermylenes by base abstraction led to insertion into Si-Si bonds and formation of silagermetes.


Cutaneous leishmaniasis in Jordan: biochemical identification of human and Psammomys obesus isolates as Leishmania major.
As part of a series of epidemiological and ecological studies of leishmaniasis in Jordan, we have made functional studies of four isolates from human lesions and from ear lesions of three field-collected Psammomys obesus. Primary isolates were subcultured, frozen stabilates prepared and BALB/c mouse infectivity experiments initiated. Each mouse was inoculated with 4-8 x 10(4) promastigotes into a hind footpad. Quantitative evaluation of the footpads showed enlargement three to four weeks postinoculation. Amastigotes were readily identified in smears from footpad lesions and promastigotes in culture. At 47 days, liver and spleen samples grew out promastigotes. Biochemical characterization of these seven isolates was made by isozyme analysis using cellulose acetate membrane electrophoresis of fructokinase, phosphoglucose isomerase, phosphoglucomutase, aspartate aminotransferase, malate dehydrogenase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. Reference isolates used for comparison were Leishmania major, L. tropica minor, L. donovani, L. aethiopica and L. m. mexicana. All seven Jordan isolates showed enzyme electromorphs identical to L. major, confirming our ecological/epidemiological studies that P. obesus is a major reservoir for human cutaneous leishmaniasis in Jordan.


SPATIAL GENETIC STRUCTURE OF CLONAL AND SEXUAL REPRODUCTION IN POPULATIONS OF ADENOPHORA GRANDIFLORA (CAMPANULACEAE).
The spatial distribution of clonal versus sexual reproduction in plant populations should generally have differing effects on the levels of biparental inbreeding and the apparent selfing rate, produced via mating by proximity through limited pollen dispersal. We used allozyme loci, join-count statistics, and Moran's spatial autocorrelation statistics to separate the spatial genetic structure caused by clonal reproduction from that maintained in sexually reproduced individuals in two populations of Adenophora grandiflora, a perennial herb. Join-count statistics showed that there were statistically significant clustering of clonal genotypes within distances less than 4 m. Both the entire populations and the sets of sexually reproduced individuals exhibited significant spatial autocorrelation at less than about 12 m, and the sexually reproduced individuals are substantially structured in an isolation-by-distance manner, consistent with a neighborhood size of about 50.


Structure of (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate reductase, the terminal enzyme of the non-mevalonate pathway.
Molecular evolution has evolved two metabolic routes for isoprenoid biosynthesis: the mevalonate and the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway. The MEP pathway is used by most pathogenic bacteria and some parasitic protozoa (including the malaria parasite, Plasmodium falciparum) as well as by plants, but is not present in animals. The terminal reaction of the MEP pathway is catalyzed by (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) reductase (LytB), an enzyme that converts HMBPP into isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Here, we present the structure of Aquifex aeolicus LytB, at 1.65 A resolution. The protein adopts a cloverleaf or trefoil-like structure with each monomer in the dimer containing three alpha/beta domains surrounding a central [Fe3S4] cluster ligated to Cys13, Cys96, and Cys193. Two highly conserved His (His 42 and His 124) and a totally conserved Glu (Glu126) are located in the same central site and are proposed to be involved in ligand binding and catalysis. Substrate access is proposed to occur from the front-side face of the protein, with the HMBPP diphosphate binding to the two His and the 4OH of HMBPP binding to the fourth iron thought to be present in activated clusters, while Glu126 provides the protons required for IPP/DMAPP formation.


Endothelial cell-binding antibodies in patients with systemic lupus erythematosus.
Abstract The implications of endothelial cell-binding IgG antibodies (EC IgG) in systemic lupus erythematosus (SLE) was evaluated by determining level of EC IgG in sera from 112 SLE patients. The serum EC IgG level was determined by the cyto-ELISA method using human umbilical vein endothelial cells (HUVEC), human microvascular endothelial cells (HMVEC), and aortic endothelial cells (HAEC) as antigens. The levels of EC IgG were significantly higher among patients with SLE than among healthy control subjects (P < 0.001), and 68% (76/112) of SLE patients were shown to be EC IgG-positive. In patients with active lupus nephritis, the level of EC IgG was statistically and significantly elevated compared with those without lupus nephritis (P < 0.05). Negative correlations between EC IgG level and levels of CH50, C3, and lymphocyte count were revealed (P < 0.05, P < 0.005, and P < 0.05, respectively). When clinical course was evaluated, the levels of EC IgG correlated with disease activity. Definitive correlations in antibody levels between HUVEC and HMVEC, and between HUVEC and HAEC were revealed (both P < 0.0001). The results of this study revealed that the EC IgG in patients with SLE was involved in the onset of clinical manifestation, especially in patients with active lupus nephritis.


Brain-targeted Near-Infrared Nanobeacon for In Situ Monitoring H2S Fluctuation during Epileptic Seizures.
Epilepsy is a neurological brain disease, and its recurrent seizures are related to the reductive substance-powered antioxidant defense system (ADS). However, until now, there has been no report on the study of in situ antioxidant fluctuation during epilepsy of varying severity. In this work, hydrogen sulfide (H2S) was selected as the model target, a H2S-responsive near-infrared fluorophore was designed and synthesized, and an amphiphilic molecule was synthesized and modified with angiopep-2 peptide at its hydrophilic terminus. A nanobeacon termed as BFPP was prepared by the formation of micelles with the package of the fluorophore. The nanobeacon was sensitive to H2S, with a low detection limit of 17 nM. The H2S fluctuation in cells can be monitored by fluorescence imaging. In addition, angiopep-2 peptide at the surface of BFPP helps it cross the blood-brain barrier, and near-infrared fluorescence improves in vivo imaging. BFPP revealed that H2S was at a moderate level in the normal brain, but its level was obviously elevated during mild epilepsy because of the activation of the ADS while significantly suppressed during severe epilepsy due to neuronal damage. This approach is generally accessible for other targets by altering the responsive fluorophore, with significance for in situ analysis of brain pathology.


Some emerging general principles in the pathogenesis of hepatocellular carcinoma.
A rational concept of the key steps in the multistep process of cancer development in liver is emerging. This concept proposes that the strategy of cancer development consists of two major sequences: (a) the genesis of persistent benign focal proliferations (clonal nodules) and (b) the development of hepatocellular cancer from one or more such nodules. Sequence (a) comprises the classical 'initiation and promotion' of chemical carcinogenesis. Sequence (b) is dominated by persistent cell proliferation. In the precancerous steps, cell proliferation is almost balanced by cell loss, leading to only a slow increase in the size of the nodules. With cancer, this balance is lost, leading to a much more rapid enlargement of the focal lesion. The carcinogenic process in the liver is viewed initially as a form of physiological adaptation to certain types of xenobiotic agents generating new focal cell populations. The animals with such new focal lesions are much better able to resist the toxic or lethal effects of many environmental hazards. According to this view, liver cancer development is a consequence of a derivative of the basic adaptation process.


Cell cycle phase perturbations and apoptosis in tumour cells induced by aplidine.
Aplidine, dehydrodidemnin B, is a marine depsipeptide isolated from the Mediterranean tunicate Aplidium albicans currently in phase II clinical trial. In human Molt-4 leukaemia cells Aplidine was found to be cytotoxic at nanomolar concentrations and to induce both a G(1) arrest and a G(2) blockade. The drug-induced cell cycle perturbations and subsequent cell death do not appear to be related to macromolecular synthesis (protein, RNA, DNA) since the effects occur at concentrations (e.g. 10 nM) in which macromolecule synthesis was not markedly affected. Ten nM Aplidine for 1 h inhibited ornithine decarboxylase activity, with a subsequently strong decrease in putrescine levels. This finding has questionable relevance since addition of putrescine did not significantly reduce the cell cycle perturbations or the cytotoxicity of Aplidine. The cell cycle perturbations caused by Aplidine were also not due to an effect on the cyclin-dependent kinases. Although the mechanism of action of Aplidine is still unclear, the cell cycle phase perturbations and the rapid induction of apoptosis in Molt-4 cells appear to be due to a mechanism different from that of known anticancer drugs.


[Investigation of the preventive effect of CoQ10 against the side-effects of anthracycline antineoplastic agents].
This study was designed to evaluate the usefulness of Coenzyme Q10 (CoQ10) in the prevention of side effects due to anthracycline agents-Adriamycin (ADM) and Daunorubicin (DNR)-by comparing the preventive effect between CoQ10-treated and non-treated groups. The subjects were 79 patients, 55 of whom had malignant lymphoma. The age range was from 16 to 77 years with a mean age of 45.4 years. CoQ10 was administered by intravenous drip at 1 mg/kg/day the day before ADM or DNR administration, on the day and for a further 2 days after administration. In mean total dose, complete remission rate and mortality, no significant differences were observed between the 2 groups. Although there were also no significant differences in the degree of alopecia, fever, nausea and vomiting, the incidences of diarrhea and stomatitis were significantly (p less than 0.10 and p less than 0.05, respectively) reduced in the CoQ10-treated group. Depression of ST waves (more than 0.05 mV) and changes in T waves (R/10 greater than T, flat, inversion) on ECG were regarded as a parameter of aggravation. Such ECG aggravation was found in 20 of 40 patients given CoQ10 (50.0%) and in 18 of 25 receiving none (72.0%); a cardiotoxicity-inhibiting tendency was thus evident (p less than 0.10). In heart rate, tachycardia was noted in the nontreated group when the period of use of anthracycline agents exceeded 8 weeks. Twenty nine patients received ADM or DNR for 8 weeks or more, and, of them, 17 were treated with CoQ10; 11 of the 17 (64.7%) showed ECG aggravation, while 11 of 12 patients (91.7%) not treated with CoQ10 showed ECG aggravation. A tendency to depress ECG aggravation was thus observed in the treated group (p less than 0.10).


[Molecular pharmacologic approaches to functional analysis of new biological target molecules for drug discovery].
This review focuses on two pharmacologic approaches to the functional evaluation of new target molecules for drug discovery. One is the development of a novel specific antagonist of the Na(+)-Ca(++) exchanger (NCX) SEA0400. The other is a comprehensive analysis of the functions of pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide ligand for G protein-coupled receptors. NCX is the one of the last target molecules regulating the cellular Ca(++) concentration. There was no efficient way to address the pathophysiologic roles of NCX until a specific antagonist, 2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline (SEA0400), was developed. Our recent studies using SEA0400 clearly showed the possible roles of NCX in several pathologic states of cardiovascular and nervous tissues. In our second approach including gene-targeting methods, we found new, unexpected roles of PACAP in higher brain functions, such as psychomotor, cognition, photoentrainment, and nociception. Based on these experimental findings, a genetic association study in schizophrenia patients revealed that the single-nucleotide polymorphisms of the PACAP gene are significantly associated with the hypofunction of the hippocampus. Regarding the peripheral roles of PACAP, we found that PACAP is involved not only in the regulation of insulin secretion in pancreatic islets, but also in the regulation of islet turnover. In subsequent phenotypic analysis of PACAP transgenic mice, we identified novel candidate genes that probably have promising functional roles.