[Clinical effect of recombinant human granulocyte colony-stimulating factor in bone marrow transplantation. The Japan rhG-CSF Clinical Study Group].
To evaluate the safety and efficacy of glycosylated recombinant human granulocyte colony-stimulating factor (rhG-CSF) derived from Chinese hamster ovary cells in bone marrow transplantation (BMT), we performed a multi-center phase II clinical study. Sixty-two bone marrow transplanted patients (46 with allograft, 16 with autograft) were enrolled in this study, and rhG-CSF was administered intravenously 30 min daily at a dose of 2, 5, 10, or 20 micrograms/kg/day for 14 consecutive days from day 1 or day 5 after transplantation. Compared with historical control, blood neutrophil recovery in greater than or equal to 5 micrograms/kg/day group was clearly accelerated without delay in other blood cells. Consistent with the acceleration of neutrophil recovery, the number of days with body temperature (greater than or equal to 38 degrees C) decreased during from day 15 to day 28. No adverse effects were observed, and there was no evidence that rhG-CSF affected the incidence of graft-versus-host disease and relapse rates of myeloid leukemia. These results suggest that the posttransplant use of rhG-CSF may be an efficient strategy for reducing the risk of bacterial and/or fungal infection complications in BMT.


Elasticity of spheres with buckled surfaces.
The buckling instabilities of core-shell systems, comprising an interior elastic sphere, attached to an exterior shell, have been proposed to underlie myriad biological morphologies. To fully discuss such systems, however, it is important to properly understand the elasticity of the spherical core. Here, by exploiting well-known properties of the solid harmonics, we present a simple, direct method for solving the linear elastic problem of spheres and spherical voids with surface deformations, described by a real spherical harmonic. We calculate the corresponding bulk elastic energies, providing closed-form expressions for any values of the spherical harmonic degree (l), Poisson ratio, and shear modulus. We find that the elastic energies are independent of the spherical harmonic index (m). Using these results, we revisit the buckling instability experienced by a core-shell system comprising an elastic sphere, attached within a membrane of fixed area, that occurs when the area of the membrane sufficiently exceeds the area of the unstrained sphere [C. Fogle et al., Phys. Rev. E 88, 052404 (2013)1539-375510.1103/PhysRevE.88.052404]. We determine the phase diagram of the core-shell sphere's shape, specifying what value of l is realized as a function of the area mismatch and the core-shell elasticity. We also determine the shape phase diagram for a spherical void bounded by a fixed-area membrane.


MicroRNA-34a Promotes Hepatic Stellate Cell Activation via Targeting ACSL1.
The incidence of liver fibrosis remains high due to the lack of effective therapies. Our previous work found that microRNA (miR)-34a expression was increased, while acy1-CoA synthetase long-chain family member1 (ACSL1) was decreased, in a dimethylnitrosamine (DNS)-induced hepatic fibrosis rat model. We hypothesized that miR-34a may play a role in the process of hepatic fibrosis by targeting ACSL1. From days 2 to 14, cultured primary hepatic stellate cells (HSCs) underwent cell morphology, immunocytochemical staining, and quantitative reverse transcription PCR (RT-qPCR) for alpha smooth muscle actin (a-SMA), desmin, rno-miR-34a, and ACSL1 expression. Wild-type and mutant luciferase reporter plasmids were constructed according to the predicted miR-34a binding site on the 3'-untranslated region (UTR) of the ACSL1 mRNA and then transfected into HEK293 cells. rno-miR-34a was silenced in HSCs to confirm that rno-miR-34a negatively regulates ACSL1 expression. mRNA and protein expression of α-SMA, type I collagen, and desmin were assayed in miR-34a-silenced HSCs. HSCs were deemed quiescent during the first 3 days and activated after 10 days. rno-miR-34a expression increased, and ACSL1 expression decreased, from day 2 to 7 to 14. rno-miR-34a was shown to specifically bind to the 3'-UTR of ACSL1. miR-34a-silenced HSCs showed higher ACSL1and lower α-SMA, type I collagen, and desmin expression than that of matching negative controls and non-transfected cells. miR-34a appears to play an important role in the process of liver fibrosis by targeting ACSL1 and may show promise as a therapeutic molecular target for hepatic fibrosis.


Membrane shaping by actin and myosin during regulated exocytosis.
The cortical actin network in neurosecretory cells is a dense mesh of actin filaments underlying the plasma membrane. Interaction of actomyosin with vesicular membranes or the plasma membrane is vital for tethering, retention, transport as well as fusion and fission of exo- and endocytic membrane structures. During regulated exocytosis the cortical actin network undergoes dramatic changes in morphology to accommodate vesicle docking, fusion and replenishment. Most of these processes involve plasma membrane Phosphoinositides (PIP) and investigating the interactions between the actin cortex and secretory structures has become a hotbed for research in recent years. Actin remodelling leads to filopodia outgrowth and the creation of new fusion sites in neurosecretory cells and actin, myosin and dynamin actively shape and maintain the fusion pore of secretory vesicles. Changes in viscoelastic properties of the actin cortex can facilitate vesicular transport and lead to docking and priming of vesicle at the plasma membrane. Small GTPase actin mediators control the state of the cortical actin network and influence vesicular access to their docking and fusion sites. These changes potentially affect membrane properties such as tension and fluidity as well as the mobility of embedded proteins and could influence the processes leading to both exo- and endocytosis. Here we discuss the multitudes of actin and membrane interactions that control successive steps underpinning regulated exocytosis.


Emerging treatment options for early mycosis fungoides.
Mycosis fungoides is a candidate for skin-directed therapies in its initial stages. In recent years, therapeutic options outside of the normal treatment recommendations such as topical imiquimod, topical tazarotene, topical methotrexate, excimer light sources, and photodynamic therapy have been published with variable results. These alternatives have been useful in cases of localized mycosis fungoides that do not respond to routine treatments; nevertheless, more studies on these methods are still needed. This article summarizes the literature and data that are known so far about these treatments.


Kainate receptor subunit diversity underlying response diversity in retinal off bipolar cells.
Postsynaptic kainate receptors mediate excitatory synaptic transmission over a broad range of temporal frequencies. In heterologous systems, the temporal responses of kainate receptors vary when different channel-forming and auxiliary subunits are co-expressed but how this variability relates to the temporal differences at central synapses is incompletely understood. The mammalian cone photoreceptor synapse provides advantages for comparing the different temporal signalling roles of kainate receptors, as cones release glutamate over a range of temporal frequencies, and three functionally distinct Off bipolar cell types receive cone signals at synapses that contain either AMPA or kainate receptors, all with different temporal properties. A disadvantage is that the different receptor subunits are not identified. We used in situ hybridization, immunocytochemistry, and pharmacology to identify the kainate receptor and auxiliary subunits in ground squirrel (Ictidomys tridecimlineatus) cb1a/b, cb2, and cb3a/b Off bipolar cell types. As expected, the types showed distinct subunit expression patterns. Kainate receptors mediated ∼80% of the synaptic response in cb3a/b cells and were heteromers of GluK1 and GluK5. Cb3a/b cells contained message for GluK1 and GluK5, and also GluK3 and the auxiliary subunit Neto1. The synaptic responses in cb1a/b cells were mediated by GluK1-containing kainate receptors that behaved differently from the receptors expressed by cb3a/b cells. AMPA receptors mediated the entire synaptic response in cb2 cells and the remaining synaptic response in cb3a/b cells. We conclude that GluK1 is the predominant kainate receptor subunit in cb1 and cb3 Off bipolar cells. Different temporal response properties may result from selective association with GluK3, GluK5, or Neto1.


Affective pattern recognition and schizophrenia.
The process by which people extract the emotional nuances of nonverbal communication is treated as a special case of pattern recognition. This process of affective pattern recognition is presumed to involve three conceptually distinct levels: The primary or cue level, the secondary or configurational level, and the propositional or predictive level. It is proposed that schizophrenia arises from defects in either or both the cue and configurational levels and that the hypothesized defects can account for the common cognitive and social-interpersonal phenomena of schizophrenia. Finally, the therapeutic implications of this model of schizophrenia are considered.


Direct analysis of hydrogen peroxide by capillary electrophoresis.
A method is described for analysis of hydrogen peroxide directly by CZE in borate buffer based on its absorption in UV light at 185 nm, without reaction with dyes. The absorption at 185 nm was about 3.5 times better than that at 214 nm. Hydrogen peroxide was generated enzymatically from glucose in aqueous solutions and in serum and was removed by the catalase enzyme. To improve the sensitivity of detection, samples were concentrated on the capillary based on stacking by ACN. The method is rapid (approximately 7 min) and specific.


Depressive symptomatology in the immediate postnatal period: identifying maternal characteristics related to true- and false-positive screening scores.
To determine whether true- and false-positive postnatal depression screening scores can be distinguished during the early postpartum period by examining characteristic differences between 2 groups: 1) women with depressive symptomatology at 1 week postpartum who continue to exhibit symptoms at 8 weeks postpartum, compared with those who do not; and 2) women with depressive symptomatology at 8 weeks postpartum who previously exhibited symptoms at 1 week postpartum, compared with those who did not. As part of a longitudinal postpartum depression study, a population-based sample of 594 women completed mailed questionnaires at 1, 4, and 8 weeks postpartum. Among women with depressive symptomatology at 1 week postpartum, diverse variables distinguished between those whose symptoms persisted or remitted at 8 weeks. These variables included recent immigrant status, psychiatric history, premenstrual symptoms, vulnerable personality, low self-esteem, child abuse history, and insufficient support. Variables that distinguished between women with depressive symptomatology at 8 weeks postpartum who previously exhibited symptoms at 1 week postpartum and those who did not included vulnerable personality, life stressors, perceived stress, insufficient support, and partner conflict. To address both the benefits and potential harms of early screening, positive screening scores on the Edinburgh Postnatal Depression Scale should also include an assessment of each individual woman's risk for postpartum depression and (or) chronic major depression.


A protocol to analyze single-cell RNA-seq data from Mycobacterium tuberculosis-infected mice lung.
Processing and analyzing single-cell RNA-seq (scRNA-seq) from lung cells are challenging due to the complexity of cell subtypes and biological variations within sample groups. Here, we present a protocol for performing an in-depth assessment on lung lymphocyte populations derived from healthy and Mycobacterium tuberculosis-infected mice. We describe steps for downloading processed scRNA-seq data, integrating samples across different conditions, and performing cluster analysis. We then detail procedures for identifying lymphoid cell subtypes, differential analysis, and pathway enrichment analysis. For complete details on the use and execution of this protocol, please refer to Akter et al. (2022).1.


Re-stratification of patients with copy-number low endometrial cancer by clinicopathological characteristics.
To stratify patients with copy-number low (CNL) endometrial cancer (EC) by clinicopathological characteristics. EC patients who underwent surgery between June 2018 and June 2022 at Peking University People's Hospital were included and further classified according to TCGA molecular subtyping: POLE ultramutated, microsatellite instability high (MSI-H), CNL, and copy-number high (CNH). Clinicopathological characteristics and prognosis of CNL patients were retrospectively reviewed. The Cox proportional hazards regression model was applied to perform univariate and multivariate analysis, and independent risk factors were identified. Differentially expressed genes (DEGs) according to overall survival (OS) were screened based on the transcriptome of CNL cases from the TCGA program. Finally, a nomogram was established, with an accuracy analysis performed. (1) A total of 279 EC patients were included, of whom 168 (60.2%) were in the CNL group. A total of 21 patients had recurrence and 6 patients deceased, and no significant difference in recurrence-free survival (RFS) was exhibited among the four molecular subtypes (P = 0.104), but that in overall survival (OS) was statistically significant (P = 0.036). (2) CNL patients were divided into recurrence and non-recurrence groups, and significant differences (P < 0.05) were found between the two groups in terms of pathological subtype, FIGO stage, ER, PR, glycated hemoglobin (HbA1c), and high-density lipoprotein cholesterol (HDL-C). All the above factors were included in univariate and multivariate Cox regression models, among which pathological subtype, PR, and HDL-C were statistically different (P < 0.05), resulting in three independent risk factors for the prognosis of patients in the CNL group. (3) By comparing the transcriptome of tumor tissues between living and deceased CNL patients from the TCGA database, 903 (4.4%) DEGs were screened, with four lipid metabolism pathways significantly enriched. Finally, a nomogram was established, and internal cross-validation was performed, showing good discrimination accuracy with an AUC of 0.831 and a C-index of 0.748 (95% CI 0.444-1.052). (4) According to the established nomogram and the median total score (85.89), patients were divided into the high score group (n = 85) and low score group (n = 83), and the 8 patients with recurrence were all in the high score group. Survival analysis was performed between the two groups, and the difference in RFS was statistically significant (P = 0.010). In the CNL group of EC patients, pathological subtype, PR, and HDL-C were independent prognostic risk factors, the nomogram established based upon which had a good predictive ability for the recurrence risk of patients with CNL EC.


High plasma levels of soluble intercellular adhesion molecule (ICAM)-1 are associated with cerebral malaria.
Cerebral malaria (CM) is responsible for most of the malaria-related deaths in children in sub-Saharan Africa. Although, not well understood, the pathogenesis of CM involves parasite and host factors which contribute to parasite sequestration through cytoadherence to the vascular endothelium. Cytoadherence to brain microvasculature is believed to involve host endothelial receptor, CD54 or intercellular adhesion molecule (ICAM)-1, while other receptors such as CD36 are generally involved in cytoadherence of parasites in other organs. We therefore investigated the contributions of host ICAM-1 expression and levels of antibodies against ICAM-1 binding variant surface antigen (VSA) on parasites to the development of CM. Paediatric malaria patients, 0.5 to 13 years were recruited and grouped into CM and uncomplicated malaria (UM) patients, based on well defined criteria. Standardized ELISA protocol was used to measure soluble ICAM-1 (sICAM-1) levels from acute plasma samples. Levels of IgG to CD36- or ICAM-1-binding VSA were measured by flow cytometry during acute and convalescent states. Wilcoxon sign rank-test analysis to compare groups revealed association between sICAM-1 levels and CM (p<0.0037). Median levels of antibodies to CD36-binding VSA were comparable in the two groups at the time of admission and 7 days after treatment was initiated (p>0.05). Median levels of antibodies to CD36-binding VSAs were also comparable between acute and convalescent samples within any patient group. Median levels of antibodies to ICAM-1-binding VSAs were however significantly lower at admission time than during recovery in both groups. High levels of sICAM-1 were associated with CM, and the sICAM-1 levels may reflect expression levels of the membrane bound form. Anti-VSA antibody levels to ICAM-binding parasites was more strongly associated with both UM and CM than antibodies to CD36 binding parasites. Thus, increasing host sICAM-1 levels were associated with CM whilst antibodies to parasite expressing non-ICAM-1-binding VSAs were not.


Factors influencing body weight one year after bariatric surgery.
The significant outcome of bariatric surgery (BS) is weight loss, which may be affected by many factors, such as initial body weight before surgery, sex, and dietary intake. Moreover, rapid weight loss is associated with an increased incidence of postsurgical cholelithiasis. To investigate the observed weight loss outcomes during the first year after BS, we investigated the factors that may influence weight loss and to detect the efficacy of prophylactic ursodeoxycholic acid against gallstone formation. This was a retrospective cohort study of all patients with morbid obesity who underwent BS in the hospital and completed a 1-year follow up. Patients with a previous BS or a history of cholecystectomy before BS were excluded. Data were extracted from the medical records at multiple postoperative intervals. There was significant weight loss in terms of percentage of excess weight loss and reduction in body mass index postoperative. A significant correlation was found between the percent of excess weight loss and age, initial body mass index, and initial weight, but there was no significant correlation with sex or type of surgery. The incidence of postoperative cholecystectomy is almost negligible. A significant association was found between age and weight loss after BS. ursodeoxycholic acid is an effective prophylaxis to decrease the incidence of cholecystectomy after BS.


The prevalence of Parkinson's disease in rural Tanzania.
The prevalence of Parkinson's disease (PD) varies worldwide from 7 to 450 per 100,000, and appears low in sub-Saharan Africa (SSA) but few data exist. We conducted a prevalence study of PD in rural Tanzania. A door-to-door study was conducted in the Hai district project area (population 161,071), using a screening questionnaire followed by structured history and examination of positive responders. Diagnosis was based on the UK PD Society Brain Bank Criteria. 33 (23 men) cases of PD were detected, with mean age 74 years (range 38-94). One patient died before the prevalence date. 78% were previously undiagnosed and untreated. Mean duration of symptoms was 5 years, and median Hoehn and Yahr stage 3. Crude prevalence rates were 30/100,000 (men), 11/100,000 (women) and 20/100,000 (combined). The direct age-standardized prevalence rates compared with the UK population were 64/100,000 (men), 20/100,000 (women) and 40/100,000 (combined). These rates are higher than previously reported from SSA, but still lower than the developed world. Many PD patients in SSA may never be diagnosed or treated, with consequent reduction in their life expectancy and quality of life. With the world population ageing PD is predicted to become an increasing problem.


Alvaxanthone, a Thymidylate Synthase Inhibitor with Nematocidal and Tumoricidal Activities.
With the aim to identify novel inhibitors of parasitic nematode thymidylate synthase (TS), we screened in silico an in-house library of natural compounds, taking advantage of a model of nematode TS three-dimensional (3D) structure and choosing candidate compounds potentially capable of enzyme binding/inhibition. Selected compounds were tested as (i) inhibitors of the reaction catalyzed by TSs of different species, (ii) agents toxic to a nematode parasite model (C. elegans grown in vitro), (iii) inhibitors of normal human cell growth, and (iv) antitumor agents affecting human tumor cells grown in vitro. The results pointed to alvaxanthone as a relatively strong TS inhibitor that causes C. elegans population growth reduction with nematocidal potency similar to the anthelmintic drug mebendazole. Alvaxanthone also demonstrated an antiproliferative effect in tumor cells, associated with a selective toxicity against mitochondria observed in cancer cells compared to normal cells.


Dentistry in Italy in the late eighteenth and early nineteenth centuries - an introduction to research.
In the late eighteenth and early nineteenth centuries the conditions of dental practice are well enough clear. Even in this period the profession continued to be practiced mostly by empirics whose training consisted of an apprenticeship carried out for varying lengths of time under some itinerant practitioner. Nevertheless in the late eighteenth century, resident dentists with stable living quarters began to appear in ever-increasing number. In this work the author gives examples of several events and advertisements who prove the just exposed assertions.


Possibilities for Implementing Fracture Liaison Service in Poland in the Light of a Visit to Glasgow Western Infirmary.
Osteoporosis remains one of the top challenges for health services all over the world. Osteoporotic changes in bone structure along with the aging of society result in rapid growth of osteoporotic fractures. Statistics show that approximately 25% of women and 20% of men will suffer a subsequent fracture within 5 years of an initial one. In order to deal with the problem, a novel program in secondary fracture prevention was developed in Scotland in the late 1990's. The system was based on a coordinator and focused on identifying, diagnosing and treating patients with osteoporotic fractures. After just a few years, the system, known as Fracture Liaison Services (FLS), proved to be a cost-effective success. For the last several years, FLS has been implemented in countries all over the world. The Glasgow Western Infirmary, where the program started, continues to be one of the top exemplary facilities in the United Kingdom. Each year the Bone Metabolism Unit proves its effectiveness by providing 4000 DXA scans and taking care of 2500 fractures a year. In 2015, the European Foundation of Osteoporosis and Musculoskeletal Diseases successfully implemented a coordinator-based Fracture Liaison Service in Poland.


Development and validation of a high performance liquid chromatography assay for the determination of temocillin in serum of haemodialysis patients.
Therapeutic drug monitoring of β-lactams can be useful for the optimization of therapy, especially when little reference data exist on actual pharmacokinetic profiles such as in patients undergoing haemodialysis. There is no reported validated method for temocillin assay in serum samples, and preliminary studies evidenced potential for interferences by acidic metabolites and co-administered drugs in patients with advanced kidney failure. This paper describes a fully validated method for the determination of temocillin in human serum, and its applicability in haemodialysis patients. Temocillin was extracted from human serum by a solid phase extraction methodology, and then assayed by reversed-phase HPLC with UV-detection. The method was validated according to the accuracy profile methodology, using total error to verify the trueness, precision and overall accuracy. It showed high specificity and precision and was accurate in the concentration range of 5-400 mg/L. LOD and LOQ were 1.2 and 5 mg/L, respectively. No interference with 30 co-administered drugs was evidenced. The method was successfully applied to clinical samples from haemodialysis patients, showing a high degree of dialysability of temocillin.


How can one be both a philosophical ethicist and a democrat?
How can one be both a philosophical ethicist and a democrat? In this article I conclude that it can be difficult to reconcile the two roles. One involves understanding, and reconciling, the conflicting views of citizens, and the other requires the pursuit of truth through reason. Nevertheless, an important function of philosophy and ethics is to inform and improve policy. If done effectively, we could expect better, and more just, laws and policies, thereby benefiting many lives. So applying philosophical thinking to policy is an important job. However, it comes with substantial difficulties, not least in reconciling, or choosing between, competing philosophical theories. Despite the importance of the task, and the apparent obstacles, there is relatively little literature on how to apply ethics to real-world policy-making. Democracies need ethicists who can engage in democratic debate and bridge the gap between philosophy and public policy. I offer some tactics here.


Embolia cutis medicamentosa.
A 35-year-old man presented with a painful skin necrosis after a deep ventrogluteal injection of diclofenac and dexamethasone for treatment of severe back pain. Immediately after the injection, the patient felt a strong pain just above the injection site. In the following days a remarkable necrosis developed in the upper gluteal region. Topical therapy with antiseptics and a topical corticosteroid cream plus analgesia with tramadole revealed no improvement of the symptoms. We excised the necrotic area. Within 1 day, the patient was without pain.


Design study of a free-electron laser on a storage ring at Tohoku University.
A free-electron laser (FEL) based on the proposed Tohoku Light Source storage ring is discussed. In the first stage the FEL is made to operate in the visible region with a relative low beam energy to avoid the complication of mirrors. Then, with a higher beam energy, the FEL can produce radiation of wavelengths in the UV or VUV region. Some simulation results of the storage-ring FEL with wavelengths of approximately 200 nm are presented.


Chloroquine-potentiating action of antihistaminics in Plasmodium falciparum in vitro.
Tricyclic antihistaminics have relatively few side-effects compared to other agents that enhance the susceptibility to chloroquine of chloroquine-resistant strains of Plasmodium falciparum. Their efficacy was tested in vitro against resistant and sensitive culture-adapted strains of P. falciparum. Isobologram analysis showed that both cyproheptadine and azatadine exert a marked synergistic action on chloroquine against chloroquine-resistant, but not against chloroquine-sensitive, strains of parasites. Cyproheptadine was about twice as effective as azatadine in reversing chloroquine resistance. Loratadine had no effect on chloroquine against both resistant and sensitive strains of parasites. Our in vitro study showed that cyproheptadine and azatadine may be promising candidates for potentiating chloroquine against resistant malaria parasites.


Laser trabeculoplasty as primary therapy in chronic open angle glaucoma.
Sixty-three phakic eyes of 54 patients with early simple (21 eyes) or capsular (42 eyes) glaucoma treated with laser trabeculoplasty (LTP) as the primary therapy were followed retrospectively 12 to 18 months after LTP. In 59 of 63 eyes followed up for 12 months and 46 of 52 eyes followed up for 18 months the intraocular pressure (IOP) was constantly maintained below 22 mmHg with no medication. After a good initial response for 6 to 12 months, 6 eyes required medical therapy because of uncontrolled pressure or progression in the damage of the optic disc or visual fields. The mean pressure reduction of all eyes 18 months after LTP was 28% in simple and 37% in capsular glaucoma. No severe complications were seen. LTP seems to be an effective and safe procedure as the primary therapy for open angle glaucoma.


A comparison of the clinical profile of Kawasaki disease in children from Northern India above and below 5 years of age.
Clinical experience collated over the last 11 years at our center suggests that Kawasaki disease (KD) affects older children more frequently as compared to the Western literature. In this study we have compared the clinical profile of KD in children above 5 years of age (Group I) with those below 5 years (Group II). Chart review of children with KD during the period January 1994-April 2006. Of the 97 children, 38 (39.2%) were in Group I and 59 (60.8%) were in Group II. Mean age at presentation of children in Group I was 8.12 +/- 2.0 years while that in Group II was 2.83 +/- 1.50 years. Mean time interval to make the diagnosis was 11.2 +/- 6.4 days in Group I as compared to 10.8 +/- 6.03 days Group II but the difference was not significant (p > 0.05). Non-purulent conjunctivitis, mucosal changes in oropharynx, cervical lymphadenopathy and hepatomegaly were seen more frequently in Group II than in Group I, but this difference was also not statistically significant. Desquamation at presentation and arthralgia/arthritis were significantly more common in Group I, while edema over hands and feet was significantly more frequent in Group II (p < 0.05). Among laboratory parameters, hemoglobin level was lower in children in Group II as compared to Group I (p = 0.001), while there was no difference in parameters of inflammation. Cardiac abnormalities were noted in about 12% children but there was no statistically significant difference between the two groups. In Chandigarh, KD occurs more frequently in children above 5 years of age as compared to the West. Older children with KD have a different clinical profile. However, cardiac complications do not differ between the 2 groups.


The effect of a beta-lactamase inhibitor peptide on bacterial membrane structure and integrity: a comparative study.
Co-administration of beta-lactam antibiotics and beta-lactamase inhibitors has been a favored treatment strategy against beta-lactamase-mediated bacterial antibiotic resistance, but the emergence of beta-lactamases resistant to current inhibitors necessitates the discovery of novel non-beta-lactam inhibitors. Peptides derived from the Ala46-Tyr51 region of the beta-lactamase inhibitor protein are considered as potent inhibitors of beta-lactamase; unfortunately, peptide delivery into the cell limits their potential. The properties of cell-penetrating peptides could guide the design of beta-lactamase inhibitory peptides. Here, our goal is to modify the peptide with the sequence RRGHYY that possesses beta-lactamase inhibitory activity under in vitro conditions. Inspired by the work on the cell-penetrating peptide pVEC, our approach involved the addition of the N-terminal hydrophobic residues, LLIIL, from pVEC to the inhibitor peptide to build a chimera. These residues have been reported to be critical in the uptake of pVEC. We tested the potential of RRGHYY and its chimeric derivative as a beta-lactamase inhibitory peptide on Escherichia coli cells and compared the results with the action of the antimicrobial peptide melittin, the beta-lactam antibiotic ampicillin, and the beta-lactamase inhibitor potassium clavulanate to get mechanistic details on their action. Our results show that the addition of LLIIL to the N-terminus of the beta-lactamase inhibitory peptide RRGHYY increases its membrane permeabilizing potential. Interestingly, the addition of this short stretch of hydrophobic residues also modified the inhibitory peptide such that it acquired antimicrobial property. We propose that addition of the hydrophobic LLIIL residues to the peptide N-terminus offers a promising strategy to design novel antimicrobial peptides in the battle against antibiotic resistance. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.


Simultaneous Native Nephrectomy and Kidney Transplantation in Patients With Autosomal Dominant Polycystic Kidney Disease.
To evaluate the feasibility of simultaneous unilateral nephrectomy with kidney transplantation and to determine the effect of this procedure on perioperative morbidity and mortality and graft and patient survival. Between January 2000 and May 2015, 145 patients with autosomal dominant polycystic kidney disease (ADPKD) underwent kidney transplantation. Of those, 40 (27.5%) underwent concurrent ipsilateral native nephrectomy (group NT). Patients in group NT were compared with patients with ADPKD not undergoing concurrent nephrectomy (group NT-) and asymptomatic patients undergoing pretransplant nephrectomy (group PNT). The average follow-up was 66 months. The graft survival rate at 1 and 5 years was 95% and 87.5% versus 93% and 76.2% in the NT and NT- groups, respectively (P = .903 and P = .544, respectively); 1-year patient survival was 100% for NT and 97% for NT- patients (P = .288), whereas 5-year patient survival was 100% and 92% for NT and NT- groups, respectively (P = .128). After propensity score matching (34 patients per group) no significant differences were observed in 1-year (97.1% in NT and 94.1%; P = 1) and 5-year (88.2% in NT and 91.2% in NT-; P = 1) graft survival, and in 1-year (100% for both groups; P = 1) and 5-year (100% in NT and 94.1% in NT-; P = 1) patient survival. Perioperative mortality was 0% among NT and 1.2% among NT- patients, whereas perioperative surgical complications were similar in both groups. One- and 5-year graft and patient survival were similar between the NT and PNT groups, but patients in the PNT group had significantly lower levels of hemoglobin and residual diuresis volumes at the time of transplant. Moreover, PNT patients had a longer pretransplant dialysis and a longer time on the waiting list. Simultaneous unilateral nephrectomy does not have a negative effect on patient and graft survival in patients with ADPKD and is associated with low morbidity. Pretransplant nephrectomy should be restricted only to highly symptomatic patients, whereas unilateral nephrectomy in asymptomatic patients should be performed during kidney transplantation only if massive kidney size precludes graft positioning.


Electrokinetic label-free screening chip: a marriage of multiplexing and high throughput analysis using surface plasmon resonance imaging.
We present an electrokinetic label-free biomolecular screening chip (Glass/PDMS) to screen up to 10 samples simultaneously using surface plasmon resonance imaging (iSPR). This approach reduces the duration of an experiment when compared to conventional experimental methods. This new device offers a high degree of parallelization not only for analyte samples, but also for multiplex analyte interactions where up to 90 ligands are immobilized on the sensing surface. The proof of concept has been demonstrated with well-known biomolecular interactant pairs. The new chip can be used for high throughput screening applications and kinetics parameter extraction, simultaneously, of interactant-protein complex formation.


Management of open lower limb injuries in South West England and Wales.
The joint British Association of Plastic, Reconstructive and Aesthetic Surgeons/British Orthopaedic Association standards define best practice management in open diaphyseal fractures of the lower limb. The aim of our study was to review the regional approach and experience in South West England and Wales. A further objective was to evaluate service provision with regard to the standards' key recommendations. A prospective audit was undertaken of open diaphyseal fracture patients. Compliance with published standards within all orthoplastic services in South West England and Wales was assessed, and facilities were evaluated. A total of 86 patients were managed between October 2012 and March 2013. This was a 56% increase from 2008. Over half (56%) presented directly to the orthoplastic services with all patients undergoing debridement within 24 hours. Two-thirds (66%) of procedures were in daylight hours excluding those requiring immediate surgical intervention. Adherence to correct antibiotic therapy was 88% at admission, 50% at primary surgery and 62% at definitive surgery. Almost two-thirds (60%) of primary procedures were performed with combined senior orthoplastic teams, with 81% achieving definitive soft tissue coverage and fixation within seven days. Compliance improved in units with larger patient caseloads and where there was an early combined approach during daylight hours. Increased open lower limb fracture workload was demonstrated across South West England and Wales, probably owing to centralisation of trauma services. An improvement in early transfer of this patient group to orthoplastic facilities has allowed all patients to be assessed and debrided within the recommended timeframe. Standards were most likely to be met in those centres seeing higher numbers of injuries and when there was a daylight hours procedure by combined orthoplastic teams.


Fatal sarin poisoning in Syria 2013: forensic verification within an international laboratory network.
During the United Nations fact-finding mission to investigate the alleged use of chemical warfare agents in the Syrian Arab Republic in 2013, numerous tissues from a deceased female victim, who had displayed symptoms of cholinergic crisis, were collected. The Organisation for the Prohibition of Chemical Weapons (OPCW) authorized two specialized laboratories in the Netherlands and Germany for forensic analysis of these samples. Diverse modern mass spectrometry (MS)-based procedures in combination with either liquid chromatography (LC) or gas chromatography (GC) separation were applied. A variety of biotransformation products of the nerve agent sarin was detected, including the hydrolysis product O-isopropyl methylphosphonic acid (IMPA) as well as covalent protein adducts with e.g., albumin and human butyrylcholinesterase (hBChE). IMPA was extracted after sample acidification by solid-phase extraction and directly analyzed by LC-tandem-MS with negative electrospray ionization (ESI). Protein adducts were found, either by fluoride-induced reactivation applying GC-MS techniques or by LC-MS-based detection after positive ESI for proteolyzed proteins yielding phosphonylated tyrosine residues or a specific phosphonylated hBChE-derived nonapeptide. These experimental results provided unambiguous evidence for a systemic intoxication and were the first proving the use of sarin in the ongoing bellicose conflict. This scenario underlines the requirement for qualified and specialized analytical laboratories to face repeated violation of the Chemical Weapons Convention.


Reduced energy expenditure after ventromedial hypothalamic lesions in female rats.
Activity, O2 consumption, and body composition were measured in female rats with lesions of the ventromedial hypothalamus (VMH). One group of lesioned rats was food restricted to maintain body weights at control levels. O2 consumption of these food-restricted VMH-lesioned rats was 13% lower than that of nonlesioned rats. About half this energy savings accrued from reduced activity; the remainder resulted from a decline in resting metabolic rate (RMR). Though matched in weight to controls, the body composition of the restricted VMH-lesioned rats was abnormal. Their carcass protein was reduced by 12%, whereas fat was elevated 200%. The replacement of lean metabolically active tissue by carcass fat appears to underlie the reduction in RMR, an interpretation supported by multiple-regression analysis of carcass composition. Ad libitum-fed VMH-lesioned rats did not show this carcass protein decline, though their fat was elevated 895%. These results confirm that reduced activity contributes to energy savings and weight gain after VMH lesions. Likewise, restricting weight of VMH-lesioned rats to control levels reduced RMR, apparently because carcass protein declines. But, since ad libitum-fed female VMH-lesioned rats do not display reduced carcass protein, the historic assumption that RMR reductions contribute to their obesity appears unwarranted.


Interdisciplinary management of an isolated intrabony defect.
The treatment of intrabony defects is a real challenge in molar teeth as it is chronic, slowly progressing disease which needs timely intervention. Periodontal inflammation associated with intrabony defect is not a separate entity as it secondarily affects the pulp causing retrograde pulpitis. However, treatment of these lesions will be complicated due to extensive bone loss. The tooth was endodontically treated followed by periodontal surgery to eliminate the deep periodontal pocket and promote bone fill in osseous defect. PepGen P-15 composited with platelet rich plasma was utilized for enhancing bone formation. The combination of these graft materials provides synergistic effect on bone regeneration.


Changes in force and stiffness during stretch of skeletal muscle fibers, effects of hypertonicity.
Slow stretch ramps (velocity: 0.17 fiber lengths s-1) were imposed during fused tetanic contractions of intact muscle fibers of the frog (1.4-3.0 degrees C; sarcomere length: 2.12-2.21 microns). Instantaneous force-extension relations were derived both under isometric conditions and during slow stretch by applying fast (0.2 ms) length steps to the fiber. An increase in tonicity (98 mM sucrose added to control Ringer solution) led to significant reduction of the maximum isometric tension but at the same time to marked increase in the force enhancement during slow stretch. The maximum force level reached during the stretch was affected very little. Experiments on relaxed fibers showed that recruitment of passive parallel elastic components were of no relevance for these effects. Hypertonicity slightly increased the instantaneous stiffness of the active fiber both in the presence and in the absence of stretch. The total extension of the undamped fiber elasticity was considerably reduced by increased tonicity under isometric conditions but was only slightly affected during slow stretch. The change in length of the undamped cross-bride elasticity upon stretch was thus greater in the hypertonic than in the normotonic solution suggesting a greater increase in force per cross-bridge in the hypertonic medium. The contractile effects are consistent with the assumptions that hypertonicity reduces the capability of the individual cross-bridge to produce active force and, furthermore, that hypertonicity has only minor effects on the number of attached cross-bridges and the maximum load-bearing capacity of the individual bridge.


Mechanisms involved in the development and healing of diabetic foot ulceration.
We examined the role of vascular function and inflammation in the development and failure to heal diabetic foot ulcers (DFUs). We followed 104 diabetic patients for a period of 18.4 ± 10.8 months. At the beginning of the study, we evaluated vascular reactivity and serum inflammatory cytokines and growth factors. DFUs developed in 30 (29%) patients. DFU patients had more severe neuropathy, higher white blood cell count, and lower endothelium-dependent and -independent vasodilation in the macrocirculation. Complete ulcer healing was achieved in 16 (53%) patients, whereas 13 (47%) patients did not heal. There were no differences in the above parameters between the two groups, but patients whose ulcers failed to heal had higher tumor necrosis factor-α, monocyte chemoattractant protein-1, matrix metallopeptidase 9 (MMP-9), and fibroblast growth factor 2 serum levels when compared with those who healed. Skin biopsy analysis showed that compared with control subjects, diabetic patients had increased immune cell infiltration, expression of MMP-9, and protein tyrosine phosphatase-1B (PTP1B), which negatively regulates the signaling of insulin, leptin, and growth factors. We conclude that increased inflammation, expression of MMP-9, PTP1B, and aberrant growth factor levels are the main factors associated with failure to heal DFUs. Targeting these factors may prove helpful in the management of DFUs.


Preaged remodeling of myofibrillar cytoarchitecture in skeletal muscle expressing R349P mutant desmin.
The majority of hereditary and acquired myopathies are clinically characterized by progressive muscle weakness. We hypothesized that ongoing derangement of skeletal muscle cytoarchitecture at the single fiber level may precede and be responsible for the progressive muscle weakness. Here, we analyzed the effects of aging in wild-type (wt) and heterozygous (het) and homozygous (hom) R349P desmin knock-in mice. The latter harbor the ortholog of the most frequently encountered human R350P desmin missense mutation. We quantitatively analyzed the subcellular cytoarchitecture of fast- and slow-twitch muscles from young, intermediate, and aged wt as well as desminopathy mice. We recorded multiphoton second harmonic generation and nuclear fluorescence signals in single muscle fibers to compare aging-related effects in all genotypes. The analysis of wt mice revealed that the myofibrillar cytoarchitecture remained stable with aging in fast-twitch muscles, whereas slow-twitch muscle fibers displayed structural derangements during aging. In contrast, the myofibrillar cytoarchitecture and nuclear density were severely compromised in fast- and slow-twitch muscle fibers of hom R349P desmin mice at all ages. Het mice only showed a clear degradation in their fiber structure in fast-twitch muscles from the adult to the presenescent age bin. Our study documents distinct signs of normal and R349P mutant desmin-related remodeling of the 3D myofibrillar architecture during aging, which provides a structural basis for the progressive muscle weakness.


[The role of phosphatidylinositol-3-kinases P85/P110 and hVPS34 in endocytosis of EGF-receptor complexes].
The previous data (Zheleznova et al., 2001) did not enable the authors to conclude which particular wortmannin sensitive PI-3-kinase--p85/p110 (I class PI-3-K) or hVPS34 (III class PI-3-K)--may be involved in the regulation of EGF-receptor endocytosis. In the present work, we have shown that upon stimulation of EGF-receptor endocytosis additional structures stained with antibody against p85 appear in A431 cells, but the p85-positive compartment never co-localized with EGF-receptor-containing compartments either in control or in wortmannin-treated cells. At the same time, wortmannin treatment prevented association of hVPS34 with endosomal membranes. We have also found that early endosomal markers--Rab5 and EEA1 (membrane association of the latter depends on Rab5 and hVPS34)--co-localized with EGF-receptor in the juxtranuclear region during late stages of endocytosis, both in control and upon wortmannin treatment. These observations favor our suggestions that the transition of EGF-receptors from early to late endosomes may occur directly in this juxtranuclear region and be tightly associated with the formation of so called multivesicular bodies (MVB), which are late endosomes per se. We suggest that wortmannin may have no effect on early EEA1-dependent stage of the receptor endocytosis but blocks a transition of EGF-receptor complexes into the late endosomes by inhibiting activity of hVPS34 and removing it from membranes. The hVPS34 product PI-3-K, according to the known data, is involved in the formation of internal vesicles of MVB. Accumulation of EGF-receptors in these vesicles is believed to be necessary for the receptor degradation.


Comparison of Etest and agar dilution method for antimicrobial susceptibility testing of Flavobacterium isolates.
The Etest was evaluated as a possible alternative to the standard agar dilution method for susceptibility testing of nine antimicrobial agents against Flavobacterium species. In studies of 100 clinical isolates, the agreement between the MICs (+/-1 log2 dilution) obtained by the two methods was acceptable for cefotaxime, ceftazidime, amikacin, minocycline, ofloxacin, and ciprofloxacin (> 90%). Conversely, the agreement between the results obtained for piperacillin was limited (84%). The overall agreement was 92.5%.


Diagnosis and surgical treatment of extrahepatic portal vein aneurysm: A case report.
Extrahepatic portal vein aneurysm is a relatively uncommon entity. We reported the case of a 51-year-old man, who was incidentally diagnosed with an asymptomatic 52 × 65 mm portal vein aneurysm, which was located at the main trunk of the portal vein, just before its bifurcation. The patient was treated with partial clamping of the portal vein aneurysm and aneurysmorrhaphy. Postoperative course was uneventful. Surgical intervention should be considered for patients with large, symptomatic or expanding portal vein aneurysms.


The effects of plaque morphological characteristics on the post-stenotic flow in left main coronary artery bifurcation.
Local post-stenotic hemodynamics has critical influence in the atherosclerotic plaque progression occurring in susceptible arterial sites, in particular the left main coronary artery (LMCA) bifurcation. Understanding the effects of plaque morphological characteristics: stenosis severity (SS), eccentricity index (EI) and lesion length (LL) on the post-stenotic flow behavior can significantly improve treatment planning. In order to investigate these effects, we have employed computational fluid dynamics (CFD) simulations in twenty computer-generated and five patient-specific LMCA models and the hemodynamic parameters: velocity, pressure (P), wall pressure gradient (WPG), wall shear stress (WSS), time averaged wall shear stress (TAWSS), oscillatory shear index (OSI), relative residence time (RRT) and helicity intensity (h2) were analyzed. Our results revealed that the effect of stenosis eccentricity varied significantly for different values of stenosis severity and lesion length. Regions with low WSS, low TAWSS and high RRT were more prominent in models having higher stenosis severity. For smaller lesion length, at low and moderate stenosis severity, surface area with low TAWSS and high RRT decreased with increasing eccentricity index, whereas for high stenosis severity models, low TAWSS region and average RRT values increased with eccentricity. However, for models with longer lesion length, regions with high OSI and RRT overall increased gradually with eccentricity. The helicity intensity (h2) of all models remained very low except at the most eccentric model with longer lesion length. The presence of very high helical flow in this model suggests the possibility of atheroprotective flow. It can be concluded that all plaque morphological characteristics covered under this investigation play an important role in plaque progression.


Cytotoxic activity of the seaweed compound fucosterol, alone and in combination with 5-fluorouracil, in colon cells using 2D and 3D culturing.
Colorectal cancer (CRC) is one of the most frequently occurring carcinomas which require effective therapies. Fucosterol is a sterol present in marine brown seaweeds with several biological activities. However, the influence of fucosterol in CRC remains to be determined. Thus, the aim of this study was to examine the anticancer activity of fucosterol alone and in combination with 5-fluorouracil (5-Fu) on two human CRC cell lines (HCT116 and HT29) and compared with cytotoxicity in one normal colon fibroblast cell line (CCD-18co) in monolayer (2D). The effect of fucosterol alone or in combination with 5-Fu was further assessed using HT29 multicellular spheroids (3D). Data demonstrated that fucosterol alone or combined with 5-Fu decreased cell viability in HT29 cells in 2D cultures without inducing cytotoxic in normal colon cells. The combination, fucosterol, and 5-Fu, also inhibited cell proliferation, clonogenic potential and cell migration without producing cell death in 2D. In multicellular spheroids, the combination fucosterol plus 5-Fu at the same concentrations used in 2D was not effective demonstrating that under the tested conditions the 3D model was more resistant than the 2D model. Taken together, these findings suggest that fucosterol might be a promising alternative to enhance the cytotoxic and anti-invasive actions of 5-Fu in colon cancer cells without consequent major adverse effects in normal cells. Our results also reinforce the need to include more complex 3D culture models in the initial stages of drug screening.


Porcine reproductive and respiratory syndrome virus Nsp1β inhibits interferon-activated JAK/STAT signal transduction by inducing karyopherin-α1 degradation.
Porcine reproductive and respiratory syndrome virus (PRRSV) inhibits the interferon-mediated antiviral response. Type I interferons (IFNs) induce the expression of IFN-stimulated genes by activating phosphorylation of both signal transducer and activator of transcription 1 (STAT1) and STAT2, which form heterotrimers (interferon-stimulated gene factor 3 [ISGF3]) with interferon regulatory factor 9 (IRF9) and translocate to the nucleus. PRRSV Nsp1β blocks the nuclear translocation of the ISGF3 complex by an unknown mechanism. In this study, we discovered that Nsp1β induced the degradation of karyopherin-α1 (KPNA1, also called importin-α5), which is known to mediate the nuclear import of ISGF3. Overexpression of Nsp1β resulted in a reduction of KPNA1 levels in a dose-dependent manner, and treatment of the cells with the proteasome inhibitor MG132 restored KPNA1 levels. Furthermore, the presence of Nsp1β induced an elevation of KPNA1 ubiquitination and a shortening of its half-life. Our analysis of Nsp1β deletion constructs showed that the N-terminal domain of Nsp1β was involved in the ubiquitin-proteasomal degradation of KPNA1. A nucleotide substitution resulting in an amino acid change from valine to isoleucine at residue 19 of Nsp1β diminished its ability to induce KPNA1 degradation and to inhibit IFN-mediated signaling. Interestingly, infection of MARC-145 cells by PRRSV strains VR-2332 and VR-2385 also resulted in KPNA1 reduction, whereas infection by an avirulent strain, Ingelvac PRRS modified live virus (MLV), did not. MLV Nsp1β had no effect on KPNA1; however, a mutant with an amino acid change at residue 19 from isoleucine to valine induced KPNA1 degradation. These results indicate that Nsp1β blocks ISGF3 nuclear translocation by inducing KPNA1 degradation and that valine-19 in Nsp1β correlates with the inhibition.


The effect of balance training intervention on postural stability in children with asthma.
Pulmonary rehabilitation is mainly focused on exercise training and breathing retraining in children with asthma. Conversely, balance training is not usually recommended for the treatment, although postural deficits were found in these patients. Therefore, this study assessed the effect of balance training intervention on postural stability in children with asthma. Nineteen children with mild intermittent asthma (age 11.1 ± 2.1 years, height 147.6 ± 13.9 cm, weight 41.8 ± 13.3 kg) were randomly assigned to an experimental group or a control group and completed a four-week physiotherapy program including breathing exercises and aerobic physical training (six times/week, 45 minutes). Both groups performed the same training, but only the experimental group underwent exercises on balance devices. The center of pressure (CoP) velocity in the anteroposterior (Vy) and mediolateral (Vx) directions, and total CoP velocity (Vtot) were recorded before and after training in the preferred and the adjusted stances under eyes open (EO) and eyes closed (EC) conditions. The addition of balance intervention led to significant improvements of Vtot (p = 0.02, p = 0.04) in both types of stance, Vx in the preferred stance (p = 0.03) and Vy in the adjusted stance (p = 0.01) under EO conditions. Significant improvements were also found in Vy in the adjusted stance (p = 0.01) under EC conditions. Results of this study support the effectiveness of balance training as a part of physiotherapy treatment for improving balance performance, predominantly under EO conditions, in children with mild asthma.


Clinical pharmacokinetics of frovatriptan.
To review available data on the clinical pharmacokinetics of frovatriptan. Preclinical data suggest that the pharmacokinetic profile of frovatriptan may differ from that of the currently available triptans. Studies of healthy volunteers, subjects with renal or hepatic impairment, elderly subjects, and patients with migraine during and between attacks were reviewed. Oral bioavailability of frovatriptan is 22% to 30%, and although the time to maximum concentration is typically 2 to 3 hours, approximately 60% to 70% of plasma maximum concentration is achieved within 1 hour of dosing. Frovatriptan distributes into erythrocytes, with binding reversible and time dependent. The relatively long terminal elimination half-life (about 26 hours) confers good systemic exposure and may produce a long duration of therapeutic action, thus reducing migraine recurrence and the need for redosing. Systemic exposure to frovatriptan generally correlates with dose between 1 and 100 mg. Blood and plasma frovatriptan concentrations are consistently higher in females, but there is no need to adjust dose according to gender. Pharmacokinetics are essentially unaffected by food and were predictable after repeat dosing; steady state is approached in about 4 to 5 days. Pharmacokinetics were changed only slightly in subjects with renal impairment or mild-to-moderate hepatic impairment, elderly individuals, and during migraine attacks. Frovatriptan is principally metabolized by the CYP1A2 isoenzyme of cytochrome P-450 and is cleared by the kidney and liver, each having sufficient capacity to compensate for impairment of the other. Frovatriptan can be taken without regard for food intake, and because of the large therapeutic margin and shallow dose-response curve, there is no need for dosage adjustment in the elderly, in women taking a combined oral contraceptive, in patients with mild-to-severe renal impairment, mild-to-moderate hepatic impairment, or according to gender. The long duration of exposure may reduce the likelihood of early migraine recurrence.


Acute undifferentiated leukaemia in a dog.
Acute undifferentiated leukaemia (AUL) is considered a separate entity in the context of acute leukaemias. AUL is extremely rare in both humans and dogs, has a rapid clinical course and does not respond to treatment. It is characterised by the presence of blast cells within the bone marrow and/or peripheral blood at levels ≥ 20% and even up to 100% of all nucleated cells. Blast cells are unable to be differentiated on morphological, cytochemical and phenotypic criteria into myeloid or lymphoid lineages because of their immaturity and/or atypia. An 8-year-old German Shepherd dog was referred for depression, asthenia, mild anaemia, thrombocytopenia and marked leucocytosis. Abdominal ultrasound showed hepatomegaly, splenomegaly, bilateral nephromegaly and enlargement of mesenteric lymph nodes. Echocardiography revealed biventricular hypertrophy with abnormal tissue density of the myocardium. Blood and bone marrow smears were composed of 95% unclassifiable and/or atypical blast cells and signs of dysplasia of the erythroid and thrombocytic/megakaryocytic lineages were present. Blast cells were negative for all cytochemical stains used and flow cytometry of peripheral blood revealed 85% of total leucocytes consisting of small-to-medium-sized cells, negative for all lymphoid and myeloid markers except CD45 and CD34. After necropsy, cytology and histology revealed that blast cells had diffusely infiltrated all tissues examined. Both erythroid and megakaryocytic extramedullary haemopoiesis was also detected in the spleen, lymph nodes and liver. All immunohistochemical stains used were negative. On the basis of all the results, a diagnosis of acute leukaemia involving a very primitive haematopoietic precursor was made.


Degradation of fibrin and elastin by intact human alveolar macrophages in vitro. Characterization of a plasminogen activator and its role in matrix degradation.
Fibrin deposition is prominent in the histopathology of a number of inflammatory lung diseases. Plasmin, activated locally in the lung, can degrade not only this fibrin but potentially structural proteins important to normal lung architecture. Because alveolar macrophages are prominent in inflammatory processes of the lung, we examined the plasminogen activator (PA) activity of human alveolar macrophages. Intact alveolar macrophages from each of 10 healthy subjects expressed PA activity. There was no difference in activity between smoking and nonsmoking individuals. The activator activity was largely cell-associated, but under certain culture conditions, macrophages released a soluble activator into the culture medium. The membrane-bound activator had an apparent molecular mass of 52-55 kD in nonreduced sodium dodecyl sulfate (SDS) gels, and monospecific antibody to urokinase neutralized the enzyme activity. Immunoprecipitation of [35S]methionine-labeled cells showed that human alveolar macrophages actually synthesize the PA in vitro. SDS-gel analysis of the immunoprecipitated material revealed the predominant species of PA to be structurally similar to reduced, active urokinase. We also examined the role of PA in the degradation of both insoluble fibrin and elastin matrices by live macrophages. Cells degraded an insoluble fibrin matrix in the presence of plasminogen whether or not the macrophages contacted the fibrin as long as proteinase inhibitors were not in the culture medium. In the presence of serum proteinase inhibitors, macrophages still degraded a fibrin matrix, but only if they were in contact with the fibrin. Live macrophages also degraded insoluble elastin only when in contact with the elastin but could do so even in the presence of serum proteinase inhibitors. In matrices containing a mixture of fibrin and elastin, cells did not degrade elastin unless plasminogen was added to the medium. These results indicate that normal alveolar macrophages synthesize and express, probably at the cell surface, a PA. The PA is physically and immunochemically similar to urokinase but is membrane bound. The PA is critical to the degradation of fibrin matrices by normal alveolar macrophages. Under tissue conditions where elastin is embedded within other structural proteins, the activator may be rate-limiting in elastin degradation as well. The findings also suggest that live macrophage proteolytic activity is relatively insensitive to the presence of serum proteinase inhibitors, suggesting a mechanism for proteolytic lung injury even in the presence of proteinase-proteinase inhibitor balance in the soluble phase.


Gut microbiome in neuropsychiatric disorders.
Neuropsychiatric disorders are a significant cause of death and disability worldwide. The mechanisms underlying these disorders include a constellation of structural, infectious, immunological, metabolic, and genetic etiologies. Advances in next-generation sequencing techniques have demonstrated that the composition of the enteric microbiome is dynamic and plays a pivotal role in host homeostasis and several diseases. The enteric microbiome acts as a key mediator in neuronal signaling via metabolic, neuroimmune, and neuroendocrine pathways. In this review, we aim to present and discuss the most current knowledge regarding the putative influence of the gut microbiome in neuropsychiatric disorders. We examined some of the preclinical and clinical evidence and therapeutic strategies associated with the manipulation of the gut microbiome. targeted taxa were described and grouped from major studies to each disease. Understanding the complexity of these ecological interactions and their association with susceptibility and progression of acute and chronic disorders could lead to novel diagnostic biomarkers based on molecular targets. Moreover, research on the microbiome can also improve some emerging treatment choices, such as fecal transplantation, personalized probiotics, and dietary interventions, which could be used to reduce the impact of specific neuropsychiatric disorders. We expect that this knowledge will help physicians caring for patients with neuropsychiatric disorders.


A General Catalytic Enantioselective Transfer Hydrogenation Reaction of β,β-Disubstituted Nitroalkenes Promoted by a Simple Organocatalyst.
Given its synthetic relevance, the catalytic enantioselective reduction of β,β-disubstituted nitroalkenes has received a great deal of attention. Several bio-, metal-, and organo-catalytic methods have been developed, which however are usually applicable to single classes of nitroalkene substrates. In this paper, we present an account of our previous work on this transformation, which implemented with new disclosures and mechanistic insights results in a very general protocol for nitroalkene reductions. The proposed methodology is characterized by (i) a remarkably broad scope encompassing various nitroalkene classes; (ii) Hantzsch esters as convenient (on a preparative scale) hydrogen surrogates; (iii) a simple and commercially available thiourea as catalyst; (iv) user-friendly procedures. Overall, the proposed protocol gives a practical dimension to the catalytic enantioselective reduction of β,β-disubstituted nitroalkenes, offering a useful and general platform for the preparation of nitroalkanes bearing a stereogenic center at the β-position in a highly enantioenriched form. A transition state model derived from control kinetic experiments combined with literature data is proposed and discussed. This model accounts and justifies the observed experimental results.


Hamuramicin C, a Cytotoxic Bicyclic Macrolide Isolated from a Wasp Gut Bacterium.
A new bicyclic macrolide, hamuramicin C (1), was isolated from Streptomyces sp. MBP16, a gut bacterial strain of the wasp Vespa crabro flavofasciata. Its 22-membered macrocyclic lactone structure was determined by NMR and mass spectrometry. The relative configurations of hamuramicin C (1) were assigned by J-based configuration analysis utilizing 1H rotating frame Overhauser effect spectroscopy and heteronuclear long-range coupling NMR spectroscopy. Genomic and bioinformatic analyses of the bacterial strain enabled identification of the type-I polyketide synthase pathway, which employs a trans-acyltransferase system. The absolute configurations of 1 were proposed based on the analysis of the sequences of ketoreductases in the modular gene cluster. Moreover, hamuramicin C (1) demonstrated significant inhibitory activity against diverse human cancer cell lines (HCT116, A549, SNU-638, SK-HEP-1, and MDA-MB-231).


Comparison of the Application of Allogeneic Fibroblast and Autologous Mesh Grafting With the Conventional Method in the Treatment of Third-Degree Burns.
Wound healing is a multipart process involving different cell types and growth factors. Third-degree burns are usually treated by early excision and skin grafting. Tissue engineering has been developed in this field in response to limitations associated with autografts. Allogeneic fibroblasts on meshed split thickness skin grafts (STSGs) are known to have useful properties in wound healing and can be used to construct a new model of living skin substitute. Fourteen patients were chosen from June 2009 until December 2010 as the sample for this study. After debridement and wound excision, meshed STSG was used to cover the entire wound. Alloskin (allofibroblasts cultured on a combination of silicone and glycosaminoglycan) was applied on one side and petroleum jelly-impregnated gauze (Iran Polymer and Petrochemical Institute) was applied on the other. The healing time, scar formation, and pigmentation score were assessed for the patients. All analyses were undertaken with SPSS 17 software. Alloskin demonstrated good properties compared to petroleum jelly-impregnated gauze. The average healing time and hypertrophic scar formation were significantly different between the two groups. In addition, the skin pigmentation score in the alloskin group was closer to normal. Alloskin grafting, including fibroblasts on meshed STSG, may be a useful method to reduce healing time and scar size and may require less autologous STSG in extensive burns where a high percentage of skin is burned and there is a lack of available donor sites.


[Dr. Wang's tube--study of a new technique for eliminating pathogens in sperm preparation for reproductive medicine].
A total of 30 ejaculates has been tested for ejaculate volume, spermatozoal concentration, motility, agglomeration, agglutination and contamination by microorganisms. After that the ejaculate has been washed twice and transferred in the Dr. Wangs Tube which was filled with medium before. After an incubation time of 90 minutes very quickly moving spermatozoan could be observed through the microscope in the upper part (C-D) of the Tube. After the Tube was cut by means of a special glasscutter the motility of the containing spermatozoans of part C-D has been determined again, followed by a second bacteriological examination of the so prepared ejaculate. The results of sperm preparation with the Dr. Wangs Tube regarding the sperm motility has been very good (85.7% of the sperm samples showed a motility rate of 90% or even more after treatment). The bacteriological examination has been satisfactory: in 77.7% of the cases all existing sperm bacterias has been eliminated by preparation in the Dr. Wangs Tube. Among other microorganisms Escherichia coli, Proteus mirabilis and Staphylococcus aureus which are considered to be pathologic could be isolated from five of the ejaculates and have been removed after treatment with the Dr. Wangs Tube. The handling of the Tube was rather demanding and Technically problematic. There has been caused glass-splinters by cutting the Tube which got visible while observing the ejaculate-medium-liquid through the microscope. Their sizes were nearly equivalent to the size of a sperms head or even bigger. If the so prepared ejaculate would be used for insemination and in-vitro fertilisation the very small splinters may destroy the success of the treatment and should be avoided or removed. The Dr. Wangs Tube offers a new modified swim-up technique for sperm preparation and is able to produce a high quality, nearly sterile sperm sample. Because of the technical deficiency of the system, the high purchase and running costs and the very small output (sperm sample volume, number of spermatozoans) the Dr. Wangs Tube is not be recommended for the reproductive medicine in its present shape.


A comparison of the in vitro cyto- and neurotoxicity of brominated and halogen-free flame retardants: prioritization in search for safe(r) alternatives.
Brominated flame retardants (BFRs) are abundant persistent organic pollutants with well-studied toxicity. The toxicological and ecological concerns associated with BFRs argue for replacement by safe(r) alternatives. Though previous research identified the nervous system as a sensitive target organ for BFRs, the (neuro) toxic potential of alternative halogen-free flame retardants (HFFRs) is largely unknown. We therefore investigated the in vitro (neuro) toxicity of 13 HFFRs and three BFRs in dopaminergic pheochromocytoma (PC12) and neuroblastoma (B35) cells by assessing several cytotoxic and neurotoxic endpoints. Effects on cell viability and production of reactive oxygen species (ROS) were measured using a combined Alamar Blue and Neutral Red assay and a H2-DCFDA assay, respectively, whereas effects on calcium homeostasis were measured using single-cell fluorescent Ca(2+)-imaging. The majority of the tested flame retardants induced negligible cytotoxicity, except zinc hydroxystannate (ZHS) and zinc stannate (ZS). A considerable fraction of flame retardants affected ROS production (decabromodiphenyl ether (BDE-209), triphenylphosphate (TPP), aluminium trihydroxide (ATH), ammonium polyphosphate (APP), magnesium hydroxide (MHO), ZHS, ZS and melamine polyphosphate (MPP)). Interestingly, ATH, ZHS, ZS and montmorillonite (MMT) increased the basal intracellular calcium concentration ([Ca(2+)]i), whereas tetrabromobisphenol A (TBBPA), resorcinol bis (diphenylphosphate) (RDP), TPP, 9,10-dihydro-9-oxa-10-phosphaphenanthrene-10-oxide (DOPO), ATH, ZHS, ZS and MMT reduced depolarization-evoked increases in [Ca(2+)]i as a result of inhibition of voltage-gated calcium channels. These combined data on the in vitro (neuro) toxicity of HFFRs in comparison with BFRs are essential for prioritization of safe(r) flame retardants. Though additional data are required for a complete (toxic) risk assessment, our data demonstrate that several HFFRs could be suitable substitutes for BFRs.


Update on neuromyelitis optica: natural history and management.
Neuromyelitis optica or Devic disease is an inflammatory disorder of the central nervous system. It is caused by antibodies that attack aquaporin 4 water channels in the cell membrane of astrocytic foot processes at the blood brain barrier. It can involve the optic nerve, the spinal cord and beyond. Here we review its pathophysiology, clinical features, and therapy.


Brooke-Spiegler syndrome with parotid gland involvement.
Salivary gland involvement in Brooke-Spiegler syndrome (BSS), an autosomal dominantly inherited disease, is known though not frequent. A case of familial cylindromatosis with parotid gland adenoma is herein reported. A 67-year-old lady presented with multiple scalp nodules and papular coalescent lesions over the nasolabial folds and the forehead. The clinical examination also revealed a left preauricular lump. Multiple biopsies of the scalp lesions and the nasolabial papules revealed cylindromas and trichoepitheliomas respectively. CT scan and FNA of the preauricular lump were suggestive of parotid gland adenoma. The patient underwent excision of the scalp cylindromas and total left parotidectomy. There is no evidence of recurrence after 4 years. The association of BSS with salivary gland tumours, emphasizes the necessity of thorough salivary gland examination in all patients with skin lesions. Knowledge of the genetic background of BSS allows for genetic counseling of patients.


The aminergic innervation of the human bronchus: a light and electron microscopic study.
Portions of bronchial wall, free of neoplasm, were obtained from human lung resected for nearby tumour. The bronchial tissue was freeze dried, fixed in formaldehyde vapour and sections examined by fluorescence microscopy. Wispy green fibres with the appearance of amine-containing nerves were seen associated with the gland acini. Fluorescent cells were also found at this location and infrequently in the epithelium. Fluorescence was blocked by pretreatment of the sections with sodium borohydride; it faded in ultraviolet light and was enhanced by incubation of the tissue with L-dopa. All these features are characteristic of specific formaldehyde-induced fluorescence of biogenic amines. Fluorescent fibres were not identified either in the smooth muscle of the airway or associated with the epithelium, although background fluorescence may have masked them at these sites. Observations by electron microscopy of human bronchial wall showed that, close to gland acini, there were nerves containing large dense-cored vesicles suggesting that they were sympathetic. After incubation of the tissue with 5-hydroxydopamine, nerves containing labelled vesicles were seen as close as 1 micron to the gland acini, though none were seen to run between the acinar cells. It is argued that these were sympathetic nerves and were close enough to the gland acini to influence their secretions. Nerves containing vesicles labelled with 5-hydroxydopamine were also seen in close association with smooth muscle cells indicating a direct sympathetic innervation of the bronchial smooth muscle. These observations are contrary to recent suggestions that the sympathetic nervous system acts only indirectly in the lungs.


Investigation of Hexylamine Adsorption on Gold in Perchloric Acid.
The adsorption of hexylamine at the solution-gold interface in 1 M HClO4 in the presence of 0.1 M Fe2+ and 0.1 Fe3+ was studied by potentiodynamic, chronoamperometric and EIS methods. The main kinetic characteristics of the oxidation-reduction reaction iron ions (exchange current density, transfer coefficient, diffusion coefficients of iron ions) were determined. It was shown that the physical adsorption of hexylamine on gold can be described by the Dhar-Flory-Huggins isotherm. The values of the adsorption constant and the Gibbs free adsorption energy were obtained. A comparison of the free adsorption energy at these interfaces with the interaction energies of hexylamine and water molecules, and hexylamine molecules with each other was carried out. It was shown that hexylamine adsorption at all of these interfaces is due mainly to the hydrophobic effect of the interaction of hexylamine and water molecules.


Determinants of performance of supplemental immunization activities for polio eradication in Uttar Pradesh, India: social mobilization activities of the Social mobilization Network (SM Net) and Core Group Polio Project (CGPP).
The primary strategy to interrupt transmission of wild poliovirus in India is to improve supplemental immunization activities (SIAs) and routine immunization coverage in priority districts. The CORE Group, part of the Social Mobilization Network (SM Net), has been successful in improving SIA coverage in high-risk areas of Uttar Pradesh (UP). The SM Net works through community level mobilisers (from the CORE Group and UNICEF) and covers more than 2 million children under the age of five. In this paper, we examine the reasons the CORE Group had been successful through exploration of which social mobilization activities of the CORE Group predicted better performance of SIAs. We carried out a secondary data analysis of routine monitoring information collected by the CORE Group and the Government of India for SIAs. These data included information about vaccination outcomes of SIAs in CORE Group areas and non-CORE Group areas within the districts where the CORE Group operates, along with information about the number of various social mobilization activities carried out for each SIA. We employed Generalized Linear Latent and Mixed Model (GLLAMM) statistical analysis methods to identify which social mobilization activities predicted SIA performance, and to account for the intra-class correlation (ICC) between multiple observations within the same geographic areas over time. The number of mosque announcements carried out was the most consistent determinant of improved SIA performance across various performance measures. The number of Bullawa Tollies carried out also appeared to be an important determinant of improved SIA performance. The number of times other social mobilization activities were carried out did not appear to determine better SIA performance. Social mobilization activities can improve the performance of mass vaccination campaigns. In the CORE Group areas, the number of mosque announcements and Bullawa Tollies carried out were important determinants of desired SIA outcomes. The CORE Group and SM Net should conduct sufficient numbers of these activities in support of each SIA. It is likely, however, that the quality of social mobilization activities (not studied here) is as or more important than the quantity of activities; quality measures of social mobilization activities should be investigated in the future as to how they determine vaccination performance.


Density-dependent shift of transforming growth factor-beta-1 from inhibition to stimulation of vascular smooth muscle cell growth is based on unconventional regulation of proliferation, apoptosis and contact inhibition.
TGF-beta shifts from inhibition to stimulation of vascular smooth muscle cell (vSMC) growth when cell density increases. How proliferation and apoptosis contribute to this shift is still unknown. In sparse and confluent V8 vSMC treated or not with TGF-beta(1) (1 ng/ml) for 3 days, cell number, mitotic activity, cell-cycle-regulatory protein levels, caspase-3 and phosphoinositide 3-kinase (PI3-K) activities were studied. In TGF-beta(1)-treated cells, (i) the growth curve rose constantly compared to controls, reaching post-confluent densities; (ii) mitotic activity, which was constant at all cell densities, was lower than in sparse but higher than in contact-inhibited control cells, and (iii) apoptosis occurred at sparse densities only. The mechanism of proliferation control by TGF-beta(1) was very unconventional in V8 vSMCs: (i) p15(INK4b) and cyclin D levels were similar in cells treated or not with TGF-beta(1), and (ii) p27(Kip1) levels remained very low even at high densities while cyclin E levels were not markedly decreased. TGF-beta(1)-induced apoptosis in sparse cultures and its reversal in dense cultures were inversely correlated to PI3-K activation. TGF-beta(1) slowed sparse V8 vSMC growth by inhibiting proliferation and inducing apoptosis. TGF-beta(1)-treated confluent vSMCs escaped contact inhibition and kept growing through unconventional regulation of p27(Kip1), cyclin E and suppression of apoptosis.


Research on the Integration of Cultural Tourism Industry Driven by Digital Economy in the Context of COVID-19-Based on the Data of 31 Chinese Provinces.
This article examines the impact of digital economy on the integration of China's cultural tourism industry in the context of COVID-19 by measuring the integration degree of cultural tourism industry as a substitute variable of cultural tourism integration. The empirical study found that the development of digital economy during the COVID-19 pandemic did promote the integration of China's cultural tourism industry, and compared with year 2019, the digital economy has strengthened the integration of cultural tourism industry. During the COVID-19 pandemic, the development of digital economy has promoted the integration of China's cultural tourism industry, and the positive effect of digital economy on the integration of China's cultural tourism industry has gradually strengthened compared to previous ones. The digital economy has played a mediating role in the impact of COVID-19 on the integration of China's cultural tourism industry. Therefore, China should formulate macropolicies and digital economy-related policies to strengthen the ability of digital economy to deal with risks and improve the digital system.


PTEN expression as a predictor for the response to trastuzumab-based therapy in Her-2 overexpressing metastatic breast cancer.
Even though trastuzumab is an effective therapy in early stage Her-2+ breast cancer, 40-50% of advanced Her-2+ breast cancer patients develop trastuzumab resistance. A potential resistance mechanism is aberrant downstream signal transmission due to loss of phosphatase and tensin homologue (PTEN). This study investigated the relationship between the expression of PTEN and trastuzumab response in Her-2 overexpressing metastatic breast cancer patients. Between 2000 and 2007, 164 patients with Her-2+ metastatic breast cancer received trastuzumab-based therapy in our institution. We analyzed PTEN status by immunohistochemistry of 115 available tumor tissues and analyzed associations with other histopathological parameters, response rate, progression free survival (PFS) and overall survival (OS) with a median follow-up of 60 months. Eighty patients were PTEN positive (69.6%) and 35 patients PTEN negative (30.4%). We found a significant association of the expression of PTEN and p53 (p = 0.041), while there was no association with grading, hormone receptor status, IGFR or MIB. We found significantly more cases with progressive disease under trastuzumab-based therapy in patients with PTEN positive breast cancers (p = 0.018), while there was no significant correlation with PFS or OS. In Her-2-positive metastatic breast cancers, PTEN positivity was significantly associated with progressive disease, but not with PFS or OS.


Origin of enantioselectivity in benzotetramisole-catalyzed dynamic kinetic resolution of azlactones.
Density functional theory (DFT) calculations were performed to investigate the origins of enantioselectivity in benzotetramisole (BTM)-catalyzed dynamic kinetic resolution of azlactones. The transition states of the fast-reacting enantiomer are stabilized by electrostatic interactions between the amide carbonyl group and the acetate anion bound to the nucleophile. The chiral BTM catalyst confines the conformation of the α-carbon and the facial selectivity of the nucleophilic attack to promote such electrostatic attractions.


Initiators of interruption in workflow: the role of MDs and RNs.
The healthcare environment has been characterized as interrupt-driven with medical doctors (MDs) and registered nurses (RNs) receiving many interruptions during a shift. Previous research studies have focused on the recipient because of the negative impact on task performance. It is equally important to understand the initiator of an interruption to help design strategies to lessen the number of interruptions and the possible negatives consequences. The purpose of this instrumental study was to examine MDs and RNs as initiators of interruptions. Results of this study indicate that MDs and RNs initiate interruptions most often through face-to-face situations and use of the telephone. Strategies to successfully manage interruptions must consider both the role of initiator as well as the recipient in an interruption event.


Dynamic changes in the appropriateness of urinary catheter use among hospitalized older patients in the emergency department.
To investigate incidence, rationales, related factors and outcomes for changing from appropriate catheter placement to inappropriate use among hospitalized older patients in the emergency department. A secondary analysis was adopted from a longitudinal study that was designed to follow the lifecycle of the urinary catheter among hospitalized older patients. Patients aged 65 and older with a urinary catheter that had been placed in the emergency department were included. Demographic factors, present health conditions, conditional factors of catheter placement, and rationales for daily urinary catheter use were collected from the original data. Inappropriate urinary catheter days were evaluated as an outcome. Appropriate urinary catheters were placed in the emergency department in 117 of the 156 patients (75%). Of these patients, 77 patients (65.8%) experienced a change from appropriate placement to inappropriate use, with a mean duration of 2.88±1.56 days. The common rationales were post-operation for hip fracture (36.3%) and no longer needing to monitor urine output (27.2%). A hierarchical regression model shows that a change from appropriate catheter placement to inappropriate use was associated with a diagnosis of urinary tract infection (OR = 0.15; 95% CI = 0.03-0.77; p = 0.02) and no record of the indication for catheter placement (OR = 4.76; 95% CI = 1.20-18.90; p = 0.02), and all variables together explained 35.9% of the variance. In addition, a change from appropriate placement to inappropriate use was further associated with prolonging inappropriate catheter-days (β = 5.34; 95% CI: 3.72-6.97; p <0.001). The study highlights a considerable percentage of change from appropriate placement to inappropriate use. Efforts to construct reminder intervention, to improve the record of catheter placement and continued attention to catheter use are necessary to reduce inappropriate urinary catheter use.


The formation of an anti-restenotic/anti-thrombotic surface by immobilization of nitric oxide synthase on a metallic carrier.
Coronary stenosis due to atherosclerosis, the primary cause of coronary artery disease, is generally treated by balloon dilatation and stent implantation, which can result in damage to the endothelial lining of blood vessels. This leads to the restenosis of the lumen as a consequence of migration and proliferation of smooth muscle cells (SMCs). Nitric oxide (NO), which is produced and secreted by vascular endothelial cells (ECs), is a central anti-inflammatory and anti-atherogenic player in the vasculature. The goal of the present study was to develop an enzymatically active surface capable of converting the prodrug l-arginine, to the active drug, NO, thus providing a targeted drug delivery interface. NO synthase (NOS) was chemically immobilized on the surface of a stainless steel carrier with preservation of its activity. The ability of this functionalized NO-producing surface to prevent or delay processes involved in restenosis and thrombus formation was tested. This surface was found to significantly promote EC adhesion and proliferation while inhibiting that of SMCs. Furthermore, platelet adherence to this surface was markedly inhibited. Beyond the application considered here, this approach can be implemented for the local conversion of any systemically administered prodrug to the active drug, using catalysts attached to the surface of the implant.


Similarity of the Escherichia coli proteome upon completion of different biopharmaceutical fermentation processes.
A comprehensive view of the physiological state of Escherichia coli cells at the completion of fermentation processes for biopharmaceutical production was attained via two-dimensional gel electrophoretic analysis of cellular proteins. For high cell density fermentations in which phosphate is depleted to induce recombinant protein expression from the alkaline phosphatase promoter, proteome analysis confirms that phosphate limitation occurs. Known phosphate starvation inducible proteins are observed at high levels; these include the periplasmic phosphate binding protein and the periplasmic phosphonate binding protein. The phn (EcoK) locus of these E. coli K-12 strains remains cryptic, as demonstrated by failure to grow with phosphonate as the sole phosphorus source. Proteome analysis also provided evidence that cells utilize alternative carbon and energy sources during these fermentation processes. To address regulatory issues in the biopharmaceutical industry, comparative electrophoretic analyses were conducted on a qualitative basis for four different fermentation processes. Using this approach, the protein profiles for these processes were found to be highly similar, with the vast majority (85-90%) of proteins detected in all profiles. The observed similarity in proteomes suggests that multiproduct host cell protein immunoassays are a feasible means of quantifying host-derived polypeptides from a variety of biopharmaceutical fermentation processes.


Alcohol consumption among Mexican American and Anglo women: results of a survey along the U.S.--Mexico border.
A household probability survey of 1233 Mexican American women and 798 Anglo women residing along the U.S.-Mexico border was conducted. A higher proportion of abstainers was found among the Mexican Americans than among the Anglos in almost every social and demographic category examined (age, marital status, education and employment status). Because the level of alcohol consumption increased markedly with the years of education completed, almost all of the overall ethnic differences observed could be accounted for by the generally lower level of education among the Mexican Americans. However, ethnic subgroups of Mexican American women reported different levels of alcohol consumption that could not be accounted for by differences in education, suggesting that additional ethnic factors contribute to drinking patterns.


Room temperature detachment nanolithography using a rigiflex polymeric mold.
We report on detachment nanolithography of an organic thin film at room temperature mediated by ultraviolet (UV) exposure. A nanopatterned, UV curable polyurethane acrylate (PUA) mold was placed on a spin-coated organic film made of 4,4'-bis[N-1-napthyl-N-phenyl-amino]biphenyl (NPB) under a low pressure (1-2 bar). A higher work of adhesion at the organic/mold interface induced detachment of the contacting layer on silicon or gold substrate, resulting in well-defined nanopatterns without a residual layer. The detachment was highly improved by a short-term UV exposure, rendering the film surface free from contaminant hydrocarbons with a lower cohesive force, as confirmed by Fourier transform infrared (FTIR) spectroscopy and measurements of contact angle of water.


Molecular quantum similarity and the fundamentals of QSAR.
A general overview on quantum similarity and applications to QSAR is presented. The concepts regarding quantum similarity from its theoretical foundation and consecutive development, involving mathematical formulation and similarity measures, are presented and complemented with application examples. The practical part, based on the well-known Crammer 31 steroids set, covers approximate quantum similarity calculations, molecular superposition, and statistics. In this way, the reader will find both basic general information and applicability of quantum similarity.


Platelet-derived growth factor stimulates bone resorption via a prostaglandin-mediated mechanism.
Platelet-derived growth factor (PDGF) stimulated up to 15-fold the production of prostaglandin E2 (PGE2) and bone resorption in neonatal mouse calvaria in organ culture. The action of PDGF on bone resorption occurred at low concentrations of the protein (ED50 = 10 ng/ml). All concentrations of PDGF which stimulated resorption also enhanced the production of PGE2 by bone; concentrations of PDGF which did not stimulate resorption did not enhance PGE2 production. PDGF-induced formation of PGE2 and bone resorption were inhibited completely by indomethacin (100 ng/ml) and hydrocortisone (1 microgram/ml). Indomethacin did not inhibit the bone resorption-stimulating activity of exogenous PGE2. In the continued presence of a maximum concentration of PDGF (100 ng/ml), an increase in bone resorption, as measured by an increase in medium calcium, was detected at 16 h (P less than 0.01), but not at 12 h; however, an increase in PGE2 production occurred within the first 2 h of treatment. A similar lag period for the onset of bone resorption was seen after the addition of exogenous PGE2 to the culture medium. On the other hand, exposure of bones to PDGF (50 ng/ml) for as brief a period as 5-15 min, followed by washout of PDGF, triggered bone resorption over the subsequent 48 h. PDGF increased cAMP production by bone within 30 min, and this effect of PDGF was blocked completely by indomethacin while the action of exogenous PGE2 on the production of cAMP was not blocked by indomethacin. The action of a low concentration of PDGF (1 ng/ml), which did not stimulate bone resorption alone, was potentiated by the phosphodiesterase inhibitor isobutylmethylxanthine (4 microM). We conclude that low concentrations of PDGF stimulate bone resorption via the enhanced local production of PGE2.


Novel cancer cell lines derived from primary breast tumors in Chinese patients.
Although many breast cancer cell lines have been used for cancer research during the past several decades, few have originated from primary tumors in Asian patients. Moreover, the incidence of breast cancer has been increasing rapidly in China during this time period. Therefore, it is essential to establish breast cancer cell lines from Chinese patients. Here, we report the establishment of three new breast cancer cell lines, designated BC-023, BC-024, and BC-034, from breast carcinoma tissues of three Chinese patients. These breast cancer cell lines grew as adherent monolayers with characteristic epithelial morphology and were maintained continuously in vitro with stable growth rates for at least 20 passages. No bacterial, fungal, or mycoplasma contamination was detected in any of the three cell lines. Additionally, these cells were human epidermal growth factor receptor 2 (C-erbB-2)-positive. All three cell lines had comparable population doubling times of 33-39 h and reproducibly formed colonies in soft agar. Furthermore, these cells displayed aggressive tumorigenicity. Thus, every characteristic of each of these cell lines meets the quality control standards of the American Type Culture Collection (ATCC). We used drug sensitivity testing with growth-inhibition assays and showed that these three lines expressed a wide range of sensitivities to cisplatin (DDP) and adriamycin (ADR). These studies indicate that these three novel cell lines may provide new models for studying cancer biology and for screening new drugs for breast cancer treatment, especially for the Chinese population.


Transcriptional Response of ATP-Binding Cassette (ABC) Transporters to Insecticide in the Brown Planthopper, Nilaparvata lugens (Stål).
The ATP-binding cassette (ABC) transporter superfamily is one of the largest groups of proteins and plays a non-negligible role in phase III of the detoxification process, which is highly involved in the response of insects to environmental stress (plant secondary metabolites and insecticides). In the present study, in Nilaparvata lugens, we identified 32 ABC transporters, which are grouped into eight subfamilies (ABCA-H) based on phylogenetic analysis. The temporal and spatial expression profiles suggested that the nymphal stages (1st-5th) and adult males showed similarity, which was different from eggs and adult females, and NlABCA1, NlABCA2, NlABCB6, NlABCD2, NlABCG4, NlABCG12, NlABCG15, and NlABCH1 were highly expressed in the midgut and Malpighian tubules. In addition, ABCG12, which belongs to the ABC transporter G subfamily, was significantly upregulated after exposure to sulfoxaflor, nitenpyram, clothianidin, etofenprox, chlorpyrifos, and isoprocarb. Moreover, verapamil significantly increased the sensitivity of N. lugens to nitenpyram, clothianidin, etofenprox, chlorpyrifos, and isoprocarb. These results provide a basis for further research on ABC transporters involved in detoxification in N. lugens, and for a more comprehensive understanding of the response of N. lugens to environmental stress.