B-Myb, a repressed trans-activating protein.
B-Myb belongs to a family of related transcription factors which share a unique DNA binding domain. B-Myb plays an important role in regulation of the cell cycle. Its expression is upregulated by the human papilloma virus HPV16 E7 oncoprotein. Overexpression of B-Myb can bypass p53-mediated cell cycle arrest. The founding member of the myb gene family, c-Myb, and A-Myb are involved in hematopoiesis and neurogenesis, respectively, and are both activators of gene transcription. Whether B-Myb is a transactivator or a repressor, however, has remained a matter of discussion. We reviewed the transactivation potential of B-Myb in yeast, taking advantage of the fact that inducible gene activation is an evolutionarily conserved process. By mutational analysis we localized a conserved activation domain in B-Myb. In vertebrate cells the transactivation potential of B-Myb is concealed by the C-terminal part of the protein. We show that the cell cycle regulators cyclin A and cyclin E activate B-Myb by eradicating the inhibition mediated by its carboxy-terminus. Our data suggest that in vertebrates the trans-activating function of B-Myb is regulated during the cell cycle and link Myb functions to cell cycle progression.


Idarubicin containing regimen in multiple myeloma: preliminary results of a pilot study using a modified "TANDEM" transplant program.
Tandem autologous transplant actually represents a challenge in multiple myeloma treatment, but the best conditioning regimen is still under investigation. With the aim of evaluating the feasibility of a modified tandem transplant strategy, we treated 10 multiple myeloma patients after conventional first line chemotherapy with a two step conditioning regimen consisting of high-dose melphalan (200 mg/m2) followed by high-dose melphalan (180 mg/m2) together with indarubicin (15 mg/sqm2 c.i. x 3 days) both with peripheral stem cell support. At first transplant, the median age wasyears, performance status was good and disease status was CR in 2 patients and PR in the rest. At the end of the first transplant, 70% of patients achieved CR and only mild toxicity was observed. After the second transplant further improvement of the response rate was obtained with 90% CR. However, we observed three toxic early infection-related deaths from CMV and legionella pneumonia at day + 17, +26, +54 after transplantation. Although this schedule seems to be effective in terms of response rate, the 30% TRM imposes an anthracycline dose-reduction with careful patient selection. This approach could reduce the toxic effects and maintain the efficacy of therapy at the same time.


Diving-related injuries in children <20 years old treated in emergency departments in the United States: 1990-2006.
The purpose of this work was to comprehensively examine diving-related injuries in the United States among children and adolescents <20 years of age. We conducted a retrospective analysis of diving-related injury data from the National Electronic Injury Surveillance System, including patients aged <20 years old who were seen in an emergency department for a diving-related injury from 1990 through 2006. An estimated 111341 patients aged <or=19 years were treated in emergency departments for diving-related injuries over the 17-year period of the study. The average annual injury rate was 8.4 injuries per 100000 US residents <20 years old. Patients aged 10 to 14 years composed the largest group (36.3%) of injured divers. Injuries to the head and/or neck (38.2%) and face (21.7%) were the most common, with the most frequent diagnoses being lacerations (33.9%) and soft tissue injuries (24.3%). Collision with a diving board and/or platform was the leading cause of injuries (43.9%). Children <10 years old had increased odds of sustaining a laceration, children <5 years old had increased odds of injury to the face, and 10- to 19-year-olds had increased odds of sustaining a fracture or an injury to the extremities. The odds of injury caused by contact with the diving board dramatically increased if the child was performing a flip and/or handstand or a backward dive. To our knowledge, this is the first study to examine recreational and competitive diving-related injuries among children and adolescents using a nationally representative sample. These results can help inform pediatricians, parents, coaches, and trainers regarding injuries seen during recreational and competitive diving and can help guide future prevention efforts.


Screening and identification of MicroRNAs expressed in perirenal adipose tissue during rabbit growth.
MicroRNAs (miRNAs) regulate adipose tissue development, which are closely related to subcutaneous and intramuscular fat deposition and adipocyte differentiation. As an important economic and agricultural animal, rabbits have low adipose tissue deposition and are an ideal model to study adipose regulation. However, the miRNAs related to fat deposition during the growth and development of rabbits are poorly defined. In this study, miRNA-sequencing and bioinformatics analyses were used to profile the miRNAs in rabbit perirenal adipose tissue at 35, 85 and 120 days post-birth. Differentially expressed (DE) miRNAs between different stages were identified by DEseq in R. Target genes of DE miRNAs were predicted by TargetScan and miRanda. To explore the functions of identified miRNAs, Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. Approximately 1.6 GB of data was obtained by miRNA-seq. A total of 987 miRNAs (780 known and 207 newly predicted) and 174 DE miRNAs were identified. The miRNAs ranged from 18 nt to 26 nt. GO enrichment and KEGG pathway analyses revealed that the target genes of the DE miRNAs were mainly involved in zinc ion binding, regulation of cell growth, MAPK signaling pathway, and other adipose hypertrophy-related pathways. Six DE miRNAs were randomly selected, and their expression profiles were validated by q-PCR. This is the first report of the miRNA profiles of adipose tissue during different growth stages of rabbits. Our data provide a theoretical reference for subsequent studies on rabbit genetics, breeding and the regulatory mechanisms of adipose development.


Nonclassical light revealed by the joint statistics of simultaneous measurements.
Nonclassicality cannot be a single-observable property, since the statistics of any quantum observable is compatible with classical physics. We develop a general procedure to reveal nonclassical behavior of light states from the joint statistics arising in the practical measurement of multiple observables. Beside embracing previous approaches, this protocol can disclose nonclassical features for standard examples of classical-like behavior, such as SU(2) and Glauber coherent states. When combined with other criteria, this would imply that every light state is nonclassical.


Gross anatomy in medical education.
The present state of human gross anatomy in medical education can generally be characterized as the presentation of a large bolus of information that is swallowed and only partly digested during the first year of medical school. The subject is often taught in a depth beyond that which would be relevant to all physicians irrespective of their future professional careers. This condition has resulted from adaptive adjustments to the escalating discrepancy between a rapidly expanding knowledge base in science and technology and the relatively fixed time period for education of a physician. Initially, traditional courses retained their comprehensive character, and new information was simply piled on top of existing departmental offerings. It soon became obvious that there would have to be a reduction in time devoted to established courses and a reciprocal expansion of time to accommodate newly developing sciences. Such adjustments were painful and often led to conflicts about what comprises essential knowledge in medical education. Thus, the curriculum time devoted to human gross anatomy has been significantly reduced to accommodate new knowledge in cellular and sub-cellular structures and other disciplines. That common foundation of knowledge, skills, values, and attitudes essential to all physicians regardless of specialty is ever-changing and often debated by medical school faculty members. However, two facts are generally agreed upon: that today's medical student with a broad but perhaps thin base in science and limited direct clinical experience is not competent upon graduation to assume patient care responsibilities without supervision and that as a result, the formal education of a physician has expanded into the graduate domain.(ABSTRACT TRUNCATED AT 250 WORDS)


Exciton localization and dissociation dynamics in CdS and CdS-Pt quantum confined nanorods: effect of nonuniform rod diameters.
One-dimensional colloidal multicomponent semiconductor nanorods, such as CdSe-CdS dot-in-rod, have been extensively studied as a promising class of new materials for solar energy conversion because of the possibilities of using the band alignment of component materials and the rod-diameter-dependent quantum confinement effect to control the location of electrons and holes and to incorporate catalysts through the growth of Pt tips. Here we used CdS nanorods as an example to study the effect of nonuniform diameters along the rod on the exciton localization and dissociation dynamics in CdS and (platinum tipped) CdS-Pt nanorods. We showed that, in CdS nanorods prepared by seeded growth, the presence of a bulb with a larger diameter around the CdS seed resulted in an additional absorption band lower in energy than the exciton in the CdS rod. As a result, excitons generated in the CdS rod could undergo ultrafast localization to the bulb region in addition to trapping on the CdS rod. We observed that the Pt tip led to fast exciton dissociation by electron transfer. However, excitons localized on the CdS bulb showed slower average ET rates than those localized in the rod region. Our findings suggested that the effect of rod morphology should be carefully considered in designing multicomponent nanorods for solar energy conversion applications.


Overexpression of CaAPX Induces Orchestrated Reactive Oxygen Scavenging and Enhances Cold and Heat Tolerances in Tobacco.
Ascorbate peroxidase (APX) acts indispensably in synthesizing L-ascorbate (AsA) which is pivotal to plant stress tolerance by detoxifying reactive oxygen species (ROS). Enhanced activity of APX has been shown to be a key step for genetic engineering of improving plant tolerance. However it needs a deeper understanding on the maintenance of cellular ROS homeostasis in response to stress. In this study, we identified and characterized an APX (CaAPX) gene from Camellia azalea. Quantitative real-time PCR (qRT-PCR) analysis showed that CaAPX was expressed in all tissues and peaked in immature green fruits; the expression levels were significantly upregulated upon cold and hot stresses. Transgenic plants displayed marked enhancements of tolerance under both cold and heat treatments, and plant growth was correlated with CaAPX expression levels. Furthermore, we monitored the activities of several ROS-scavenging enzymes including Cu/Zn-SOD, CAT, DHAR, and MDHAR, and we showed that stress tolerance was synchronized with elevated activities of ROS-scavenging. Moreover, gene expression analysis of ROS-scavenging enzymes revealed a role of CaAPX to orchestrate ROS signaling in response to temperature stresses. Overall, this study presents a comprehensive characterization of cellular response related to CaAPX expression and provides insights to breed crops with high temperature tolerances.


Purification, crystallization and characterization of N-acetylneuraminate lyase from Escherichia coli.
N-Acetylneuraminate lyase produced by Escherichia coli was purified and crystallized from a genetically engineered strain (E. coli SF8/pNAL1). The enzyme showed apparent molecular masses of 105,000 Da on gel filtration and 35,000 Da on SDS/PAGE, suggesting that the enzyme is a trimer. The apparent optimum pH and temperature were found to be 6.5-7.0 and 80 degrees C respectively. The Km values for N-acetylneuraminate and N-glycollylneuraminate were 3.3 and 3.3 mM respectively. The enzyme was inhibited by reduction with NaBH4 in the presence of the substrate, indicating that the enzyme belongs to the Schiff-base-forming Class I aldolases. The enzyme was strongly inhibited by Cu2+ ions, p-chloromercuribenzoate and N-bromosuccinimide, and also inhibited competitively by the reaction product, pyruvate, and its structurally related compounds, dihydroxyacetone and DL-glyceraldehyde.


A bipartite yeast SSRP1 analog comprised of Pob3 and Nhp6 proteins modulates transcription.
The FACT complex of vertebrate cells, comprising the Cdc68 (Spt16) and SSRP1 proteins, facilitates transcription elongation on a nucleosomal template and modulates the elongation-inhibitory effects of the DSIF complex in vitro. Genetic findings show that the related yeast (Saccharomyces cerevisiae) complex, termed CP, also mediates transcription. The CP components Cdc68 and Pob3 closely resemble the FACT components, except that the C-terminal high-mobility group (HMG) box domain of SSRP1 is not found in the yeast homolog Pob3. We show here that Nhp6a and Nhp6b, small HMG box proteins with overlapping functions in yeast, associate with the CP complex and mediate CP-related genetic effects on transcription. Absence of the Nhp6 proteins causes severe impairment in combination with mutations impairing the Swi-Snf chromatin-remodeling complex and the DSIF (Spt4 plus Spt5) elongation regulator, and sensitizes cells to 6-azauracil, characteristic of elongation effects. An artificial SSRP1-like protein, created by fusing the Pob3 and Nhp6a proteins, provides both Pob3 and Nhp6a functions for transcription, and competition experiments indicate that these functions are exerted in association with Cdc68. This particular Pob3-Nhp6a fusion protein was limited for certain Nhp6 activities, indicating that its Nhp6a function is compromised. These findings suggest that in yeast cells the Cdc68 partners may be both Pob3 and Nhp6, functioning as a bipartite analog of the vertebrate SSRP1 protein.


The spectrum of cytomegalovirus infection following human heart-lung transplantation.
Data were analyzed from 19 long-term survivors of cardiopulmonary transplantation in this institution, including nine patients with normal pulmonary function and 10 recipients with posttransplant obliterative bronchiolitis. In all cases, donor cytomegalovirus titers (IgG), preoperative recipient titers (IgG), and serial postoperative recipient titers (IgM, IgG, and complement fixation) were available. In addition, surveillance cytomegalovirus cultures and pulmonary function tests were obtained prospectively after surgery in all 19 patients. A total of 12 patients developed active cytomegalovirus infection (serologic conversion confirmed by positive cultures) after transplantation, six of whom subsequently developed obliterative bronchiolitis. However, infection was clinically associated with pulmonary deterioration in only four of these patients, three of whom had cytomegalovirus pneumonitis. With the exception of obliterative bronchiolitis, no other permanent sequelae of cytomegalovirus infection were evident in this small group. Progressive obliterative bronchiolitis was also seen in four of the seven recipients who had no evidence of cytomegalovirus infection at any time. Although viral causes have been associated with obliterative bronchiolitis, the current data suggest that cytomegalovirus infection in the absence of pneumonitis does not appear to be a significant risk factor for obliterative bronchiolitis in cardiopulmonary transplant recipients. A larger group of patients will be required to ultimately establish the role of cytomegalovirus infection in this setting.


Pituitary adenomas that produce adrenocorticotropic hormone and alpha-subunit: clinicopathological, immunohistochemical, ultrastructural, and immunoelectron microscopic studies in nine cases.
Eight surgical and one autopsy specimen of pituitary adenomas (six cases of Cushing's disease, two of Nelson's syndrome, and one of hypopituitarism) were studied by histochemical, immunohistocytological, and ultrastructural methods. Eight tumors showed the characteristic histochemical profile of corticotroph adenoma--amphophilic to basophilic, and periodic acid-Schiff-positive to some extent. In all tumors, immunohistochemical studies revealed adrenocorticotropic hormone (ACTH) and alpha-subunit in the cytoplasm of some adenoma cells. By electron microscopy, seven tumors were found to be monomorphous; six were typical corticotroph adenomas and one was a subtype II silent corticotroph adenoma. One unique lesion was bimorphous--i.e., composed of corticotrophs as well as cells resembling glycoprotein cells. Immunoelectron microscopy by the double-labeling immunogold technique, performed on one corticotroph adenoma, demonstrated the presence of ACTH and alpha-subunit not only within the same adenoma cells but also within the same secretory granules. The cytogenesis of ACTH alpha-subunit tumors, a rare form of plurihormonal adenoma, remains to be elucidated. The duration of disease associated with these tumors exceeded the duration in patients with ordinary corticotroph adenomas. Given the low frequency with which increases in serum alpha-subunit are detectable in patients with such tumors--13% in this series--hormone production is not recognized at preoperative evaluation.


Sub-femtosecond time-resolved measurements of electron bunches with a C-band radio-frequency deflector in x-ray free-electron lasers.
Time-resolved diagnostics are fundamental for x-ray free-electron lasers (FELs). Radio-frequency (RF) transverse deflector structures (TDSs) are typically employed to characterize the temporal properties of the electron beams driving FELs. In this article, we present time-resolved measurements with a resolution below one femtosecond using a C-band RF TDS at SwissFEL, the x-ray FEL facility at the Paul Scherrer Institute in Switzerland. The sub-femtosecond resolution is partially achieved due to an optimized optics setup and fits the expected values, showing a good understanding of our models. Measurements with a sub-femtosecond resolution are of crucial importance for ultra-fast x-ray FEL applications.


Open globe injuries with positive intraocular cultures: factors influencing final visual acuity outcomes.
To investigate the clinical features influencing final visual acuity outcomes of eyes with positive intraocular cultures after open globe injuries. Retrospective, consecutive, interventional case series. Thirty-seven patients. The medical records were reviewed of all patients with positive intraocular cultures after open globe injuries treated at Bascom Palmer Eye Institute between January 1, 1995, and December 31, 2001. Final visual acuity. Clinical features investigated included the following: (1). presence or absence of clinical endophthalmitis; (2). virulence of the cultured organism (coagulase-negative Staphylococci, Corynebacterium, and Propionibacterium acnes were classified as nonvirulent organisms, whereas all other organisms were classified as virulent organisms); (3). presence of intraocular foreign body (IOFB); (4). presence of retinal detachment; (5). interval between ocular injury and surgical repair; (6). severity of vision loss at presentation; (7). zone of injury; (8). wound length; and (9). presence of vitreous hemorrhage. The study included 37 eyes of 37 patients with a mean age of 30 years (range, 18 months-85 years) and a median follow-up of 13 months (range, 1-71 months). Study eyes were stratified into two groups: group 1 eyes (n = 16) were those in which clinical endophthalmitis did not develop, whereas group 2 eyes (n = 21) were those in which clinically diagnosed endophthalmitis developed at some point during their clinical course. Presenting visual acuity was similar in the two groups (mean logarithm of the minimum angle of resolution [logMAR] acuity, 1.91 and 2.22 [Snellen equivalents, 2/162 and 2/331] respectively; P = 0.33). Final acuities in the two groups were different, but not to a statistically significant level (mean logMAR acuity, 1.14 and 2.05 [Snellen equivalents, 20/276 and 2/224], respectively; P = 0.069). In group 1, final visual acuity ranged from 20/20 to no light perception (median acuity, 20/186); 12 eyes (75.0%) achieved a final visual acuity of 20/400 or better. In group 2, final visual acuity ranged from 20/25 to no light perception (median acuity, 7/200); of 20 eyes with known final visual acuity, 10 (50.0%) retained 20/400 or better vision. In group 1, three eyes (19%) eyes had virulent organisms, and 13 eyes (81%) had nonvirulent organisms. In group 2, 12 eyes (57%) had virulent organisms, and nine eyes (43%) had nonvirulent organisms. A final acuity of 20/60 or better was achieved in 14 eyes (41%), and a final acuity of 20/400 or better was achieved in 22 eyes (59%). Better presenting visual acuity (P = 0.038), culture of a nonvirulent organism (P = 0.011), lack of a retinal detachment (P = 0.002), absence of clinical endophthalmitis (P = 0.069), and shorter wound length (P = 0.024) were associated with better visual acuity outcome. In four of six eyes (67%) with both an IOFB and clinical endophthalmitis (group 2), the final visual acuity was no light perception (IOFB was not itself significantly associated with final visual acuity; P = 0.11). Among eyes with positive intraocular cultures after open globe injury, the visual prognosis is guarded. Clinical features associated with better visual acuity outcomes include better presenting visual acuity, culture of a nonvirulent organism, lack of a retinal detachment, absence of clinical endophthalmitis, and shorter wound length.


Childhood optic neuritis clinical features and outcome.
To describe clinical features and outcome of a series of children with first-episode optic neuritis investigated in three paediatric neurology centres. Databases were searched to identify children (<16 years) with optic neuritis and life table analysis was used. 44 children (female/male ratio 1.8) median age 10.9 years were followed up for median 1 year. Optic neuritis was unilateral in 43%. Maximal visual deficit was severe (<6/60) in 77%, with full recovery in 70%. Cumulative probability of developing MS (11/44) or NMO (3/44) at 2 years was 0.45. Relapsing optic neuritis was a strong predictor for development of MS or NMO. A positive MRI (>1 brain T2 hyperintense lesion) was a strong predictor for development of MS. Childhood optic neuritis is associated with severe visual deficit with good recovery. An initial abnormal MRI brain scan or relapsing optic neuritis should alert the clinician to MS or NMO diagnosis.


Quantitative PCR assays to detect whales, rockfish, and common murre environmental DNA in marine water samples of the Northeastern Pacific.
Monitoring aquatic species by identification of environmental DNA (eDNA) is becoming more common. To obtain quantitative eDNA datasets for individual species, organism-specific quantitative PCR (qPCR) assays are required. Here, we present detailed methodology of qPCR assay design and testing, including in silico, in vitro, and in vivo testing, and comment on the challenges associated with assay design and performance. We use the presented methodology to design assays for three important marine organisms common in the California Current Ecosystem (CCE): humpback whale (Megaptera novaeangliae), shortbelly rockfish (Sebastes jordani), and common murre (Uria aalge). All three assays have excellent sensitivity and high efficiencies ranging from 92% to 99%. However, specificities of the assays varied from species-specific in the case of common murre, genus-specific for the shortbelly rockfish assay, and broadly whale-specific for the humpback whale assay, which cross-amplified with other two other whale species, including one in a different family. All assays detected their associated targets in complex environmental water samples.


Herbal medicine AnoSpray suppresses proinflammatory cytokines COX-2 and RANTES in the management of hemorrhoids, acute anal fissures and perineal wounds.
Hemorrhoids, anal fistula and fissure are common anorectal complications. Anorectal diseases are associated with severe pain, inflammation, swelling, itching and bleeding. These diseases may be managed with different medical treatments or surgical procedures, depending on their severity. Surgical procedures, however, are highly invasive and are associated with higher costs and the possibility of recurrence. In addition, surgical removal of fistula-in-ano leads to the formation of perineal wounds. Therefore, developing therapeutic interventions that are effective in alleviating inflammation and pain are desirable for the effective management of anorectal diseases. Herbal compounds have previously been indicated to suppress inflammation and pain in different pathological conditions. The aim of the present study was to examine the effects elicited by a polyherbal formulation, AnoSpray®, on the migration of inflammatory cells and on the expression of inflammatory cytokines in anorectal diseases. The effect of AnoSpray on cell viability and migration was studied using MTT and wound-migration assays, respectively. Furthermore, the effects of AnoSpray on the expression of the inflammatory cytokines regulated upon activation, normal T cell expressed and presumably secreted (RANTES) and VEGF, as well as on cyclooxygenase-2 (COX)-2, were investigated using western blot analysis. The expression of RANTES and COX-2 in human hemorrhoid specimens was also analyzed to corroborate the in vitro findings. The results obtained revealed that AnoSpray did not exhibit any cytotoxic effects; however, it did lead to a significant suppression in the migration of RAW 264.7 and BJ cells. Furthermore, the results suggested that AnoSpray suppressed the expression of the inflammatory cytokines RANTES and VEGF, and also the expression of COX-2. In addition, RANTES and COX-2 were significantly downregulated in the clinical specimens of AnoSpray-treated hemorrhoids compared with the controls. Taken together, the results of the present study suggested that AnoSpray may be a potential therapeutic agent in the treatment of bleeding hemorrhoids, anal fissures and perineal wounds.


High Levels of Psychological Resilience Are Associated With Decreased Anxiety in Inflammatory Bowel Disease.
Anxiety and depression are comorbid disorders with IBD and are associated with poor outcomes. Resilience is an innate but modifiable trait that may improve the symptoms of psychological disorders. Increasing resilience may decrease the severity of these comorbid disorders, which may improve IBD outcomes. The aim of this study was to describe the association between resilience, anxiety, and depression in IBD patients. We performed a cross-sectional study of IBD patients. Patients completed a questionnaire consisting of the Connor-Davidson Resilience Scale (CD-RISC), a measure of resilience, the Generalized Anxiety Disorder 7 (GAD-7), and the Patient Health Questionnaire-9. Primary outcome was severity of anxiety and depression in patients with high resilience. Multivariable linear regression analysis evaluated the association between severity of anxiety and depression and level of resilience. A sample of 288 patients was analyzed. Bivariable linear regression analysis showed a negative association between resilience and anxiety (Pearson rho = -0.47; P < .0001) and between resilience and depression (Pearson rho = -0.53; P < .0001). Multivariable linear regression indicated that high resilience is independently associated with lower anxiety and that for every 1-unit increase in CD-RISC, the GAD-7 score decreased by 0.04 units (P = .0003). Unlike anxiety, the association between resilience and depression did not remain statistically significant on multivariable analysis. High resilience is independently associated with lower anxiety in IBD patients, and we report a quantifiable decrease in anxiety score severity for every point of increase in resilience score. These findings suggest that IBD patients with higher resilience may have better coping mechanisms that buffer against the development of anxiety.


Micro-spatial variation of soil metal pollution and plant recruitment near a copper smelter in Central Chile.
Soil chemical changes produced by metal smelters have mainly been studied on a large scale. In terms of plant survival, determination of small scale variability may be more important because less toxic microhabitats may represent safe sites for successful recruitment and thus for plant survival. Three dominant microhabitats (open spaces and areas below the canopy of Sphaeralcea obtusiloba and Baccharis linearis shrubs) were defined in a heavily polluted area near a copper smelter and characterised in terms of microclimate, general soil chemistry, total and extractable metal concentrations in the soil profile (A0 horizon, 0-5 and 15-20 cm depth), and seedling densities. Results indicated a strong variability in microclimate and soil chemistry not only in the soil profile but also among microhabitats. Air/soil temperatures, radiation and wind speed were much lower under the canopy of shrubs, particularly during the plant growth season. Soil acidification was detected on top layers (0-5 cm depth) of all microhabitats while higher concentrations of N, Cu and Cd were detected on litter and top soil layers below shrubs when compared to open spaces; however, high organic matter content below shrubs decreased bioavailability of metals. Plant recruitment was concentrated under shrub canopies; this may be explained as a result of the nursery effect exerted by shrubs in terms of providing a more favourable microclimate, along with better soil conditions in terms of macronutrients and metal bioavailability.


Prevalence of rectal, urethral, and pharyngeal chlamydia and gonorrhea detected in 2 clinical settings among men who have sex with men: San Francisco, California, 2003.
The Centers for Disease Control and Prevention developed screening and diagnostic testing guidelines for chlamydia and gonorrhea at urethral, rectal, and pharyngeal sites for men who have sex with men (MSM). However, in most clinical settings, rectal chlamydial testing is not performed for MSM, and primarily sexually transmitted disease (STD) clinics alone perform routine rectal and pharyngeal gonorrhea screening for asymptomatic men. We evaluated the prevalence of rectal, urethral, and pharyngeal chlamydial and gonococcal infections among MSM seen at the municipal STD clinic and the gay men's community health center. We also determined the proportion of asymptomatic rectal infections, described the patterns of single and multiple anatomic sites of infection, and evaluated the proportion of chlamydial infections that would be missed and not treated if MSM were not routinely tested for chlamydia. We tested specimens using previously validated nucleic acid amplification tests (NAATs). The prevalence of infection varied by anatomic site (chlamydia: rectal, 7.9%; urethral, 5.2%; and pharyngeal, 1.4%; for gonorrhea, rectal, 6.9%; urethral, 6.0%; and pharyngeal, 9.2%). Approximately 85% of rectal infections were asymptomatic supporting the need for routine screening. Because 53% of chlamydial infections and 64% of gonococcal infections were at nonurethral sites, these infections would be missed and not treated if only urethral screening was performed. In addition, >70% of chlamydial infections would be missed and not treated if MSM were tested only for gonorrhea. Because these infections enhance both HIV transmission and susceptibility, clinical settings serving MSM should evaluate the prevalence of chlamydial and gonococcal infections by anatomic site using validated NAATs.


Oral administration of a curcumin-phospholipid formulation (Meriva®) for treatment of chronic diabetic macular edema: a pilot study.
The purpose of this open-label study was to investigate the effect of a curcumin-phospholipid lecithin formulation (Meriva®) on visual acuity and optical coherence tomography (OCT) retinal thickness in patients with chronic diabetic macular edema. Curcumin-phospholipid lecithin formulation (Meriva®, Indena S.p.A, Milan, Italy) was administered as tablets (Norflo®, Eye Pharma, Genoa, Italy) twice a day. Visual acuity and macular edema as measured by OCT before and after curcumin-phospholipid formulation treatment were assessed. The study included 12 eyes from 11 patients who completed at least a 3-month follow-up period. After 3 months of therapy, no eyes showed reduction in visual acuity, 16% showed stabilization, and 84% showed improvement. The improvement was statistically significant (p = 0.0072). After 3 months of therapy, 92% of eyes showed reduction of macula edema, 8% showed stabilization, and 0% showed an increase (p = 0.009). Our results, albeit preliminary, suggest that a curcumin-phospholipid formulation (Meriva®), administered as Norflo® tablets, may be feasible in the improvement of visual acuity and reduction of macular edema in patients with diabetic retinopathy.


Solid-Phase Extraction Approaches for Improving Oligosaccharide and Small Peptide Identification with Liquid Chromatography-High-Resolution Mass Spectrometry: A Case Study on Proteolyzed Almond Extract.
Reverse-phase solid-phase extraction (SPE) is regularly used for separating and purifying food-derived oligosaccharides and peptides prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. However, the diversity in physicochemical properties of peptides may prevent the complete separation of the two types of analytes. Peptides present in the oligosaccharide fraction not only interfere with glycomics analysis but also escape peptidomics analysis. This work evaluated different SPE approaches for improving LC-MS/MS analysis of both oligosaccharides and peptides through testing on peptide standards and a food sample of commercial interest (proteolyzed almond extract). Compared with conventional reverse-phase SPE, mixed-mode SPE (reverse-phase/strong cation exchange) was more effective in retaining small/hydrophilic peptides and capturing them in the high-organic fraction and thus allowed the identification of more oligosaccharides and dipeptides in the proteolyzed almond extract, with satisfactory MS/MS confirmation. Overall, mixed-mode SPE emerged as the ideal method for simultaneously improving the identification of food-derived oligosaccharides and small peptides using LC-MS/MS analysis.


Risk factors for community-acquired Escherichia coli bacteraemia: a systematic review protocol.
Introduction: Rates of community-acquired Escherichia coli bacteraemia (ECB) have been consistently rising. As rates of antimicrobial resistance (AMR), particularly in Gram-negative bacteria, are also increasing, this is of concern both for management of individual patients and healthcare systems. There is currently little data on the risk factors for development of community-acquired ECB: this review aims to identify these risk factors in order to inform community interventions to reduce ECB as well as antibiotic prescribing policy. Methods and analysis: We will search Medline (Ovid), Embase (Ovid), Web of Science/Scopus and the Cochrane Central Register of Controlled Trials for published reports on observational and experimental primary research studies involving patients admitted to hospital with community-acquired ECB. Two reviewers will independently screen the studies for eligibility, perform data collection and assess study quality and risk of bias. Random effects meta-analyses will be performed if appropriate. Ethics and dissemination: No primary data will be collected for this study and so formal ethical approval is not required. We will publish the results of our review in relevant peer-reviewed medical journals, and will also seek to present them at relevant medical conferences. PROSPERO registration number: CRD42018104402.


A divide-and-conquer approach to determine the Pareto frontier for optimization of protein engineering experiments.
In developing improved protein variants by site-directed mutagenesis or recombination, there are often competing objectives that must be considered in designing an experiment (selecting mutations or breakpoints): stability versus novelty, affinity versus specificity, activity versus immunogenicity, and so forth. Pareto optimal experimental designs make the best trade-offs between competing objectives. Such designs are not "dominated"; that is, no other design is better than a Pareto optimal design for one objective without being worse for another objective. Our goal is to produce all the Pareto optimal designs (the Pareto frontier), to characterize the trade-offs and suggest designs most worth considering, but to avoid explicitly considering the large number of dominated designs. To do so, we develop a divide-and-conquer algorithm, Protein Engineering Pareto FRontier (PEPFR), that hierarchically subdivides the objective space, using appropriate dynamic programming or integer programming methods to optimize designs in different regions. This divide-and-conquer approach is efficient in that the number of divisions (and thus calls to the optimizer) is directly proportional to the number of Pareto optimal designs. We demonstrate PEPFR with three protein engineering case studies: site-directed recombination for stability and diversity via dynamic programming, site-directed mutagenesis of interacting proteins for affinity and specificity via integer programming, and site-directed mutagenesis of a therapeutic protein for activity and immunogenicity via integer programming. We show that PEPFR is able to effectively produce all the Pareto optimal designs, discovering many more designs than previous methods. The characterization of the Pareto frontier provides additional insights into the local stability of design choices as well as global trends leading to trade-offs between competing criteria.


[Methods of laboratory studies in the etiology of acute epididymitis].
A laboratory study in order to establish etiology of acute epididymitis (AE) was made in 243 patients. The complex of etiological diagnosis included: microscopic investigation of urethral discharge or urine, bacteriological examination of an initial urine portion and wound discharge in the operated on patients. In addition, urethral discharge/urine was studied with polymerase chain reaction (PCR) in 96 patients; in 46 men PCR examined operative material. It was found that in sexually active young patients AE was caused primarily by infectious agents of sexually transmitted diseases. Targeted etiotropic therapy results in clinical cure in the majority of the cases. In older patients the disease was of bacterial origin (gram-negative bacteria, as a rule, are causative agents) and in some cases is associated with development of pyodestructive epidydimitis.


Effects of noise and proficiency on intelligibility of Chinese-accented English.
This study compared the intelligibility of native and foreign-accented bilingualism English speech presented in quiet and mixed with three different levels of background noise. Two native American English speakers and four native Mandarin Chinese speakers for whom English is a second language each read a list of 50 phonetically balanced sentences (Egan, 1948). The authors speech intelligibility identified two of the Mandarin-accented English speakers as high-proficiency speakers and two as lower proficiency speakers, based on their speech intelligibility in quiet (about 95% and 80%, respectively). Original record-perception ings and noise-masked versions of 48 utterances were presented to monolingual American English speakers. Listeners were asked to write down the words they heard the speakers say, and intelligibility was measured as content words correctly identified. While there was a modest difference between native and high-proficiency speech in quiet (about 7%), it was found that adding noise to the signal reduced the intelligibility of high-proficiency accented speech significantly more than it reduced the intelligibility of native speech. Differences between the two groups in the three added noise conditions ranged from about 12% to 33%. This result suggests that even high-proficiency non-native speech is less robust than native speech when it is presented to listeners under suboptimal conditions.


Photoinduced superwetting single-walled carbon nanotube/TiO(2) ultrathin network films for ultrafast separation of oil-in-water emulsions.
A SWCNT/TiO2 nanocomposite ultrathin film that has superhydrophilic and underwater superoleophobic properties after UV-light irradiation is successfully prepared by coating TiO2 via the sol-gel process onto an SWCNT ultrathin network film. The robust and flexible SWCNT/TiO2 nanocomposite films with a thickness and pore size of tens of nanometers can separate both surfactant-free and surfactant-stabilized oil-in-water emulsions in an ultrafast manner with fluxes up to 30 000 L m(-2) h(-1) bar(-1), which is 2 orders of magnitude higher than commercial filtration membranes with similar separation performance, and with high separation efficiency. Most importantly, the films exhibit excellent antifouling and self-cleaning performance during multiple cycles with the aid of the photocatalytic property of TiO2 nanoparticles. This work provides a route for designing ultrathin and superwetting films for effective separation of oil-in-water emulsions. The SWCNT/TiO2 ultrathin film is potentially useful in treating emulsified wastewater produced in industry and daily life and for purification of crude oil and fuel.


Perceived dental treatment need among older Tanzanian adults - a cross-sectional study.
Need perceptions for dental care play a key role as to whether people in general will seek dental care. The aim was to assess the prevalence of perceived need of problem based dental care, dental check-ups and any type of dental care. Guided by the conceptual model of Wilson and Cleary, the relationship of perceived need for dental care with socio-demographic characteristics, clinically defined dental problems and self-reported oral health outcomes was investigated. Partial prosthetic treatment need was estimated using a socio-dental approach. A cross-sectional survey was conducted in Pwani region and in Dar es Salaam in 2004/2005. Information from interviews and clinical examination became available for 511 urban and 520 rural adults (mean age 62.9 yr). 51.7% (95% CI 46.2, 57.0) urban and 62.5 % (95% CI 53.1, 70.9) rural inhabitants confirmed need for dental check-up, 42.9% (95% CI 36.9, 48.9) urban and 52.7% (95% CI 44.5, 60.6) rural subjects confirmed need for problem oriented care and 38.4% (95% CI 32.4, 44.6) urban versus 49.6% (95% CI 41.8, 57.4) rural residents reported need for any type of dental care. Binary and ordinal multiple logistic regression analyses revealed that adults who reported bad oral health and broken teeth were more likely to perceive need for dental care across the three outcome measures than their counterparts. Socio-demographic factors and clinically defined problems had less impact. Based on a normative and an integrated socio-dental approach respectively 39.5% and 4.7% were in need for partial dentures. About half of the participants confirmed need for problem oriented care, dental check-ups and any type of dental care. Need perceptions were influenced by perceived oral health, clinically assessed oral problems and socio-demographic characteristics. Need estimates for partial denture was higher when based on clinical examination alone compared to an integrative socio-dental approach.


Loss of coproporphyrin by centrifuging urine samples before liquid-chromatography.
We have previously reported a "high-performance" liquid-chromatographic procedure for urinary porphyrins. While attempting to automate this procedure, we discovered that coproporphyrin could be partly or completely lost from urine samples if they were centrifuged or filtered before analysis. The loss is related to the amount of precipitate in each sample, acidic urines showing the greatest losses. If samples are alkalinized before centrifugation, the loss is prevented.


[Insufficient justification for granting a wrongful-life claim].
In 2005 the Dutch Supreme Court granted a so-called 'wrongful-life' claim. Such a claim can only be sustained by establishing that it would have been better for the child not to have been born at all. Because the Court refrained from establishing this, similar claims involving less severe handicaps may unfortunately be filed.


Evaluating the Far-Infrared Radiation Bioeffects on Micro Vascular Dysfunction, Nervous System, and Plantar Pressure in Diabetes Mellitus.
The most frequent clinical complication is diabetes. Diabetes is characterized by elevated blood glucose levels resulting in sensory nerve damage or lesions. Diabetic foot wounds are often slow to heal and require medical attention and monitoring. This study evaluates the effect of far-infrared radiation on the microcirculation and plantar pressure in the diabetic foot. Ten diabetics and 4 nondiabetics were recruited in this study. The diabetic group was examined before and after the intervention in each month for 3 consecutive months. Four nondiabetic groups were also measured before and after the intervention for 2 weeks in each month. The surface temperature and blood flow in the diabetic foot was significantly improved (temperature: 32.1 ± 2.3°C vs 33.5 ± 2.2°C, P < .05; blood flow image: 118.3 ± 58.1 PU [perfusion unit] vs 50.4 ± 4.3 PU, P < .05). The sympathetic nerve activity index LF also increased from 40.8 ± 18.6% to 61.8 ± 13.5% (P = .07) in the second month. Plantar pressure tended to increase in the third month. This might indicate that far-infrared radiation could affect the mechanical properties of the plantar foot soft tissue. These results indicated that the effects of far-infrared radiation would improve blood circulation and change the soft tissue properties in the diabetic foot.


Two years into COVID-19 - Lessons in SARS-CoV-2 and a perspective from papers in FEBS Letters.
The 2019 outbreak of coronavirus disease (COVID-19) in Wuhan (Hubei province of China) has given rise to a pandemic spread of virus, more than 240 million incidences and a death toll larger than 5 million people. COVID-19 has set off large efforts in research, therapy and patient care, as well as public and private debates in every imaginable form. A number of scientists used the publication platforms provided by the Federation of the European Biochemical Societies (FEBS) to present their research data, reviews, opinions and other contributions relating to COVID-19 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, I highlight the recent COVID-19 papers which have been published and collected in a Virtual Issue in FEBS Letters, and discuss their implications towards understanding the molecular, biochemical and cellular mechanisms of SARS-CoV-2 infections, vaccine development and antiviral discovery strategies.


Relationships between Cerebral Vasculopathies and Microinfarcts in a Community-Based Cohort of Older Adults.
Cerebral microinfarcts are associated with cognitive impairment and dementia. Small vessel diseases such as cerebral arteriolosclerosis and cerebral amyloid angiography (CAA) have been found to be associated with microinfarcts. Less is known about the associations of these vasculopathies with the presence, numbers, and location of microinfarcts. These associations were examined in the clinical and autopsy data of 842 participants in the Adult Changes in Thought (ACT) study. Both vasculopathies were categorized by severity (none, mild, moderate, and severe) and region (cortical and subcortical). Odds ratios (OR) and 95% CIs for microinfarcts associated with arteriolosclerosis and CAA adjusted for possible modifying covariates such as age at death, sex, blood pressure, APOE genotype, Braak, and CERAD were estimated. 417 (49.5%) had microinfarcts (cortical, 301; subcortical, 249), 708 (84.1%) had cerebral arteriolosclerosis, 320 (38%) had CAA, and 284 (34%) had both. Ors (95% CI) for any microinfarct were 2.16 (1.46-3.18) and 4.63 (2.90-7.40) for those with moderate (n = 183) and severe (n = 124) arteriolosclerosis, respectively. Respective Ors (95% CI) for the number of microinfarcts were 2.25 (1.54-3.30) and 4.91 (3.18-7.60). Similar associations were observed for cortical and subcortical microinfarcts. Ors (95% Cis) for the number of microinfarcts associated with mild (n = 75), moderate (n = 73), and severe (n = 15) amyloid angiopathy were 0.95 (0.66-1.35), 1.04 (0.71-1.52), and 2.05 (0.94-4.45), respectively. Respective Ors (95% Cis) for cortical microinfarcts were 1.05 (0.71-1.56), 1.50 (0.99-2.27), and 1.69 (0.73-3.91). Respective Ors (95% Cis) for subcortical microinfarcts were 0.84 (0.55-1.28), 0.72 (0.46-1.14), and 0.92 (0.37-2.28). These findings suggest a significant association of cerebral arteriolosclerosis with the presence, number, and location (cortical and subcortical) of microinfarcts, and a weak and non-significant association of CAA with each microinfarct, highlighting the need for future research to better understand the role of small vessel diseases in the pathogenesis of cerebral microinfarcts.


Agents in development for the management of cocaine abuse.
Cocaine abuse is a serious health problem in many areas of the world, yet there are no proven effective medications for the treatment of cocaine dependence. Preclinical studies suggest that the reinforcing effect of cocaine that promotes its abuse is mediated by blockade of the presynaptic dopamine transporter. This results in increased dopamine activity in the mesolimbic or meso-accumbens dopamine reward system of brain. Development of new medications to treat cocaine dependence has focused on manipulation of this dopamine system, either by direct action on dopamine binding sites (transporter or receptors) or indirectly by affecting other neurotransmitter systems that modulate the dopamine system. In principle, a medication could act via one of three mechanisms: (i) as a substitute for cocaine by producing similar dopamine effects; (ii) as a cocaine antagonist by blocking the binding of cocaine to the dopamine transporter; or (iii) as a modulator of cocaine effects by acting at other than the cocaine binding site. The US National Institute on Drug Abuse has a Clinical Research Efficacy Screening Trial (CREST) programme to rapidly screen existing medications. CREST identified four medications warranting phase II controlled clinical trials: cabergoline, reserpine, sertraline and tiagabine. In addition, disulfiram and selegiline (deprenyl) have been effective and well tolerated in phase II trials. However, selegiline was found ineffective in a recent phase III trial. Promising existing medications probably act via the first or third aforementioned mechanisms. Sustained-release formulations of stimulants such as methylphenidate and amfetamine (amphetamine) have shown promise in a stimulant substitution approach. Disulfiram and selegiline increase brain dopamine concentrations by inhibition of dopamine-catabolising enzymes (dopamine-beta-hydroxylase and monoamine oxidase B, respectively). Cabergoline is a direct dopamine receptor agonist, while reserpine depletes presynaptic stores of dopamine (as well as norepinephrine and serotonin). Sertraline, baclofen and vigabatrin indirectly reduce dopamine activity by increasing activity of neurotransmitters (serotonin and GABA) that inhibit dopamine activity. Promising new medications act via the second or third aforementioned mechanisms. Vanoxerine is a long-acting inhibitor of the dopamine transporter which blocks cocaine binding and reduces cocaine self-administration in animals. Two dopamine receptor ligands that reduce cocaine self-administration in animals are also undergoing phase I human safety trials. Adrogolide is a selective dopamine D(1) receptor agonist; BP 897 is a D(3) receptor partial agonist.A pharmacokinetic approach to treatment would block the entry of cocaine into the brain or enhance its catabolism so that less cocaine reached its site of action. This is being explored in animals using the natural cocaine-metabolising enzyme butyrylcholinesterase (or recombinant versions with enhanced capabilities), catalytic antibodies, and passive or active immunisation to produce anti-cocaine binding antibodies. A recent phase I trial of a "cocaine vaccine" found it to be well tolerated and producing detectable levels of anti-cocaine antibodies for up to 9 months after immunisation.


The usefulness of immunohistochemistry in the diagnosis of follicular lymphoma in bone marrow biopsy specimens.
We used a panel of paraffin antibodies to determine whether neoplastic and nonneoplastic lymphoid aggregates in the bone marrow can be distinguished reliably. Formalin-fixed, paraffin-embedded bone marrow core biopsy specimens with lymphoid aggregates were stained using primary antibodies directed against bcl-2, bcl-6, CD5, CD10, CD20, and CD23. We studied 61 cases (26 follicular lymphoma and 35 benign or atypical aggregates). We found that no single stain is sufficient for identification of neoplastic lymphoid aggregates. However, this distinction was made possible by using a panel of antibodies. Under the conditions we tested, the most useful antibodies were CD10, bcl-2, CD5, and CD20. Most benign or atypical aggregates do not express CD10 and CD23. In addition, nonneoplastic aggregates had a large population of T cells. bcl-2 was useful in an architectural context for distinguishing neoplastic aggregates. bcl-6 often was expressed in both neoplastic and nonneoplastic aggregates and, thus, poorly discriminated between these processes. We studied the expression of CD10 and bcl-6 in selected lymph nodes in some cases.


Dystrophin Deficiency Leads to Genomic Instability in Human Pluripotent Stem Cells via NO Synthase-Induced Oxidative Stress.
Recent data on Duchenne muscular dystrophy (DMD) show myocyte progenitor's involvement in the disease pathology often leading to the DMD patient's death. The molecular mechanism underlying stem cell impairment in DMD has not been described. We created dystrophin-deficient human pluripotent stem cell (hPSC) lines by reprogramming cells from two DMD patients, and also by introducing dystrophin mutation into human embryonic stem cells via CRISPR/Cas9. While dystrophin is expressed in healthy hPSC, its deficiency in DMD hPSC lines induces the release of reactive oxygen species (ROS) through dysregulated activity of all three isoforms of nitric oxide synthase (further abrev. as, NOS). NOS-induced ROS release leads to DNA damage and genomic instability in DMD hPSC. We were able to reduce both the ROS release as well as DNA damage to the level of wild-type hPSC by inhibiting NOS activity.


CO electroxidation on gold in alkaline media: a combined electrochemical, spectroscopic, and DFT study.
The aim of the present work is to provide a deeper understanding of gold catalysis for CO electrooxidation in alkaline media, through a combined electrochemical, spectroscopic, and DFT study. Voltammetric and spectroscopic measurements evidence that the amount of CO irreversibly adsorbed on gold increases as the adsorption potential becomes more negative (vs SHE). This explains why higher CO coverages can be achieved in more alkaline solutions, since the value of adsorption potential vs RHE becomes more negative vs SHE with increasing pH. On the other hand, the combination of FTIRRAS experiments and DFT calculations shows that the adsorption site of irreversibly adsorbed CO on Au(111) depends on the value of the adsorption potential. It is concluded that CO adsorption on top sites takes place at all studied potentials, and hollow and bridge sites also become occupied for adsorption potentials lower and higher than 0 V vs RHE, respectively. However, it should be noted that our DFT calculations give values of the CO binding energies that are not strong enough to explain CO irreversible adsorption. This may be partly attributed to the fact that OH coadsorption is not included in the calculations. Indeed, this work presents two experimental facts that suggest that CO adsorption on gold promotes the coadsorption of OH species: (i) CO irreversibly adsorbed on Au(111) and Au(100) leads to an unusual voltammetric feature, whose charge indicates the stabilization of one OH species per adsorbed CO species; (ii) the apparent transfer coefficient of this unusual state is close to unity, suggesting that it is due to a presumed structural transformation coupled to OH adsorption. Finally, the effect of the adsorption potential on the bulk CO electrooxidation is also studied. It is found that, on Au(111), an increased occupation of CO on multifold (hollow) sites seems to result in a less efficient catalysis. However, on Au(110), an increased coverage of CO on top sites does not produce any significant change in catalysis.


Effect of fatty acids on action of polyene antibiotics.
Fatty acids cause a decrease in the absorption spectra of the antifungal polyene macrolide antibiotics nystatin, filipin, candicidin, and amphotericin B. For nystatin, filipin, and candicidin, this decrease in absorption could be correlated with the activity of the fatty acids in protecting the yeast Saccharomyces cerevisiae against the action of these antibiotics. With amphotericin B a correlation was observed between the decrease in absorption caused by certain derivatives of fatty acids and the protective action of these derivatives against the activity of amphotericin B on yeast. It is concluded that, like the sterols, fatty acids also interact with the polyene antibiotics and thereby reduce their effective concentrations.


Structural, functional, and metabolic correlates in ischemic hearts: effects of substrates.
The ultrastructure, function, and metabolism of isolated rat hearts perfused under control or ischemic conditions were investigated. Either both glucose (11 mM) or glucose and palmitic acid (1.5 mM) were used as metabolic substrates. A 60-min period of whole-heart ischemia, i.e., a 60% initial reduction in coronary flow, resulted in a more dramatic morphological alteration in those hearts receiving palmitate compared to those receiving glucose as the only substrate. In ischemic hearts receiving palmitate, intramitochondrial osmiophilic amorphous densities of both rounded and elongated types were observed. These amorphous densities did not develop in ischemic hearts receiving glucose alone over the same period of ischemia. Such morphological alterations were associated with a more severe deterioration of mechanical function in the presence of palmitate. Both ischemic conditions resulted in increased tissue levels of acyl esters of CoA and carnitine, but the rise in levels of long-chain acyl carnitine was about two times greater in those ischemic hearts receiving palmitate.


A novel species-specific PCR assay for identifying Lactobacillus fermentum.
Lactobacillus fermentum is a Gram-positive bacterium that is associated with active caries lesions. Methods for identifying Lactobacillus species traditionally have been based upon culture methods coupled with biochemical tests, which are generally unreliable. The aim of this study was to develop a species-specific PCR assay for the direct detection of L. fermentum in oral clinical samples. PCR primers specific for L. fermentum were identified by alignment of bacterial 16S rRNA genes and selection of sequences specific for L. fermentum at their 3' ends. PCR positivity for L. fermentum DNA was indicated by amplification of a 337 bp product. The primers were shown to be specific for L. fermentum DNA, since no PCR product was obtained when genomic DNA from a wide range of other oral bacteria, including closely related Lactobacillus species, were used as test species. The PCR assay was then used in an attempt to identify L. fermentum DNA in supragingival plaque samples and in pus aspirates from subjects with acute dento-alveolar abscesses. Four out of 70 (5.7 %) supragingival plaque samples analysed were positive for the presence of L. fermentum DNA while none of the 19 pus samples analysed was positive for L. fermentum DNA. This PCR assay provides a more rapid, specific and sensitive alternative to conventional culture methods for the identification of L. fermentum in clinical specimens.


The Role of Natural Killer Cells in Autoimmune Diseases.
Natural killer (NK) cells, the large granular lymphocytes differentiated from the common lymphoid progenitors, were discovered in early 1970's. They are members of innate immunity and were initially defined by their strong cytotoxicity against virus-infected cells and by their important effector functions in anti-tumoral immune responses. Nowadays, NK cells are classified among the recently discovered innate lymphoid cell subsets and have capacity to influence both innate and adaptive immune responses. Therefore, they can be considered as innate immune cells that stands between the innate and adaptive arms of immunity. NK cells don't express T or B cell receptors and are recognized by absence of CD3. There are two major subgroups of NK cells according to their differential expression of CD16 and CD56. While CD16+CD56dim subset is best-known by their cytotoxic functions, CD16-CD56bright NK cell subset produces a bunch of cytokines comparable to CD4+ T helper cell subsets. Another subset of NK cells with production of interleukin (IL)-10 was named as NK regulatory cells, which has suppressive properties and could take part in immune-regulatory responses. Activation of NK cells is determined by a delicate balance of cell-surface receptors that have either activating or inhibitory properties. On the other hand, a variety of cytokines including IL-2, IL-12, IL-15, and IL-18 influence NK cell activity. NK-derived cytokines and their cytotoxic functions through induction of apoptosis take part in regulation of the immune responses and could contribute to the pathogenesis of many immune mediated diseases including ankylosing spondylitis, Behçet's disease, multiple sclerosis, rheumatoid arthritis, psoriasis, systemic lupus erythematosus and type-1 diabetes. Dysregulation of NK cells in autoimmune disorders may occur through multiple mechanisms. Thanks to the rapid developments in biotechnology, progressive research in immunology enables better characterization of cells and their delicate roles in the complex network of immunity. As NK cells stand in between innate and adaptive arms of immunity and "bridge" them, their contribution in inflammation and immune regulation deserves intense investigations. Better understanding of NK-cell biology and their contribution in both exacerbation and regulation of inflammatory disorders is a requisite for possible utilization of these multi-faceted cells in novel therapeutic interventions.


CD44/chondroitin sulfate proteoglycan and alpha 2 beta 1 integrin mediate human melanoma cell migration on type IV collagen and invasion of basement membranes.
Tumor cell invasion of basement membranes (BM) represents one of the critical steps in the metastatic process. Tumor cell recognition of individual BM matrix components may involve individual cell adhesion receptors, such as integrins or cell surface proteoglycans, or may involve a coordinate action of both types of receptors. In this study, we have focused on the identification of a cell surface CD44/chondroitin sulfate proteoglycan (CSPG) and alpha 2 beta 1 integrin on human melanoma cells that are both directly involved in the in vitro invasion of reconstituted BM via a type IV collagen-dependent mechanism. Interfering with cell surface expression of human melanoma CSPG with either p-nitro-phenyl-beta-D-xylopyranoside treatment or anti-CD44 monoclonal antibody (mAb) preincubation (mAb) preincubation inhibits melanoma cell invasion through reconstituted BM. These treatments also strongly inhibit melanoma cell migration on type IV collagen, however, they are ineffective at inhibiting cell adhesion to type IV collagen. Purified melanoma cell surface CD44/CSPG, or purified chondroitin sulfate, bind to type IV collagen affinity columns, consistent with a role for CD44/CSPG-type IV collagen interactions in mediating tumor cell invasion. In contrast, melanoma cell migration on laminin (LM) does not involve CD44/CSPG, nor does CD44/CSPG bind to LM, suggesting that CD44/CSPG-type IV collagen interactions are specific in nature. Additionally, anti-alpha 2 and anti-beta 1 integrin mAbs are capable of blocking melanoma cell invasion of reconstituted BM. Both of these anti-integrin mAbs inhibit melanoma cell adhesion and migration on type IV collagen, whereas only anti-beta 1 mAb inhibits cell adhesion to LM. Collectively, these results indicate that melanoma cell adhesion to type IV collagen is an important consideration in invasion of reconstituted BM in vitro, and suggest that CD44/CSPG and alpha 2 beta 1 integrin may collaborate to promote human melanoma cell adhesion, migration, and invasion in vivo.


Prognosis of Synchronous Colorectal Liver Metastases After Simultaneous Curative-Intent Surgery According to Primary Tumor Location and KRAS Mutational Status.
Tumor location and KRAS mutational status have emerged as prognostic factors of colorectal cancer. We aimed to define the prognostic impact of primary tumor location and KRAS mutational status among synchronous colorectal liver metastases (CRLM) patients who underwent simultaneous curative-intent surgery (SCIS). We compared the clinicopathologic characteristics and long-term outcomes of 227 patients who underwent SCIS for synchronous CRLM, according to tumor location and KRAS mutational status. We cross-classified tumor location and KRAS mutational status and compared survival outcomes between the four resulting patient groups. Forty-one patients (18.1%) had right-sided (RS) tumors and 186 (81.9%) had left-sided (LS) tumors. One-third of tumors (78/227) harbored KRAS mutations. The KRAS mutant-type (KRAS-mt) was more commonly observed among RS tumors than among LS tumors [21/41 (51.2%) vs. 57/186 (30.6%), p = 0.012]. Median follow-up time was 43.4 months. Patients with RS tumors had shorter survival times than those with LS tumors [median disease-free survival (DFS): RS, 9.9 months vs. LS, 12.1 months, p = 0.003; median overall survival (OS): RS, 49.7 months vs. LS, 88.8 months, p = 0.039]. RS tumors were a negative prognostic factor for DFS [hazard ratio (HR) 1.878, p = 0.001] and OS (HR 1.660, p = 0.060). RS KRAS-mt and LS KRAS wild-type (KRAS-wt) tumors had the worst and best oncological outcomes, respectively. Tumor location has a prognostic impact in patients who underwent SCIS for CRLM, and RS KRAS-mt tumors yielded the worst oncological outcome. These results may allow for more tailored multimodality treatments.


Rivaroxaban Plus Aspirin Versus Aspirin Alone After Endovascular Revascularization for Symptomatic PAD: Insights From VOYAGER PAD.
Rivaroxaban plus aspirin compared with aspirin alone reduced major cardiac and ischemic limb events after lower extremity revascularization (LER) in the VOYAGER PAD (Vascular Outcomes Study of ASA Along With Rivaroxaban in Endovascular or Surgical Limb Revascularization for Peripheral Artery Disease) trial. The effect has not been described in patients undergoing endovascular LER. The VOYAGER PAD trial randomized 6564 patients with symptomatic peripheral artery disease to a double-blinded treatment with 2.5 mg of rivaroxaban BID or matching placebo and 100 mg of aspirin daily. The primary efficacy outcome was a composite of acute limb ischemia, major amputation of a vascular pathogenesis, myocardial infarction, ischemic stroke, or cardiovascular death. The principal safety end point was Thrombolysis in Myocardial Infarction major bleeding. A prespecified subgroup of patients who underwent endovascular revascularization was included. Endovascular LER occurred in 4379 (66.7%) patients and surgical LER in 2185 (33.3%). Over a 3-year follow-up, rivaroxaban reduced the risk of the primary outcome by 15% (hazard ratio [HR], 0.85 [95% CI, 0.76-0.96]) with an absolute risk reduction of 0.92% at 6 months and 1.04% at 3 years and a consistent benefit in those receiving endovascular (HR, 0.89 [95% CI, 0.76-1.03]) or surgical LER (HR, 0.81 [95% CI, 0.67-0.98]; P interaction=0.43). For endovascular-treated patients, rivaroxaban reduced the risk of acute limb ischemia or major amputation of a vascular pathogenesis by 30% (HR, 0.70 [95% CI, 0.54-0.90]; P=0.005) with an absolute risk reduction of 1.0% at 6 months and 2.0% at 3 years compared with aspirin alone. Among endovascular-treated patients, the median duration of concomitant dual antiplatelet therapy with clopidogrel treatment was 31 days (interquartile range, 30-58). There was a consistent benefit for rivaroxaban regardless of background clopidogrel. Thrombolysis in Myocardial Infarction major bleeding was significantly higher for the rivaroxaban and aspirin group for the endovascular cohort (HR, 1.66 [95% CI, 1.06-2.59]) with an absolute risk increase of 0.9% at 3 years with no increase in intracranial or fatal bleeding observed (HR, 0.86 [95% CI, 0.40-1.87]; P=0.71). Mortality with rivaroxaban was higher in the endovascular-treated patients (HR, 1.24 [95% CI, 1.02-1.52]), although this finding was isolated to specific regions. Rivaroxaban added to aspirin or dual antiplatelet therapy after LER for peripheral artery disease reduces ischemic risk and increases major bleeding without an increased risk of intracranial or fatal bleeding. These benefits are consistent in those treated with endovascular and surgical approaches with significant benefits for major adverse limb events. These data support the use of rivaroxaban in addition to aspirin or dual antiplatelet therapy after endovascular intervention for symptomatic peripheral artery disease.


Antibiotic associated diarrhoea: infectious causes.
Nearly 25% of antibiotic associated diarrhoeas (AAD) is caused by Clostridium difficile, making it the commonest identified and treatable pathogen. Other pathogens implicated infrequently include Clostridium perfringens, Staphylococcus aureus, Klebsiella oxytoca, Candida spp. and Salmonella spp. Most mild cases of AAD are due to non-infectious causes which include reduced break down of primary bile acids and decrease metabolism of carbohydrates, allergic or toxic effects of antibiotic on intestinal mucosa and pharmacological effect on gut motility. The antibiotics most frequently associated with C. difficile associated diarrhoea are clindamycin, cephalosporin, ampicillin and amoxicillin. Clinical presentation may vary from mild diarrhoea to severe colitis and pseudomembranous colitis associated with high morbidity and mortality. The most sensitive and specific diagnostic test for C. difficile infection is tissue culture assay for cytotoxicity of toxin B. Commercial ELISA kits are available. Though less sensitive, they are easy to perform and are rapid. Withdrawal of precipitating antibiotic is all that is needed for control of mild to moderate cases. For severe cases of AAD, oral metronidazole is the first line of treatment, and oral vancomycin is the second choice. Probiotics have been used for recurrent cases.


A review of continuing professional development for dentists in Europe.
To summarise findings from a literature review of dentists' engagement in continuing professional development (CPD) and its effects on improving oral health care for patients. The search strategy used key terms in a range of databases and an academic literature search engine, complemented by hand searching and citation follow-up. One hundred and fourteen papers were reviewed. The majority of dentists engaged in CPD. Factors affecting participation included time since graduation, costs, work and home commitments, postgraduate qualification, interest and convenience. Learning needs identification and reflection on practice were rarely evidenced. Common modes of CPD were courses and journal reading; no one delivery method proved more effective. Few papers directly explored recommendations for topics although suggestions related to common areas of error and gaps in knowledge or skill. Studies of CPD effectiveness and impact-on-practice suggested that courses can result in widespread new learning and considerable self-reported change in practice. However, significant barriers to implementing change in workplace practice were noted and included availability of materials, resources and support from colleagues. To ensure high standards of care, alongside recommending core or mandatory topics, more attention should be given to reflection on learning needs, the learner's readiness to engage with education and training and the influence of the workplace environment.


Refined avian risk assessment for aldicarb in the United States.
Aldicarb was recently reviewed by the US Environmental Protection Agency (USEPA) for re-registration eligibility. In this paper, we describe a refined avian risk assessment for aldicarb that was conducted to build upon the screening-level methods used by USEPA. The goal of the refined ERA was to characterize and understand better the risks posed by aldicarb to birds in areas where the pesticide is applied. Aldicarb is a systemic insecticide sold in granular form under the trade name Temik. It is applied directly to soil and is used to control mites, nematodes, and aphids on a variety of crops (e.g., cotton, potatoes, peanuts). Consumption of grit is necessary for proper digestion in many bird species, particularly for granivores and insectivores. Thus, aldicarb granules may be mistaken for grit by birds. The Granular Pesticide Avian Risk Assessment Model (GranPARAM) is described in a companion paper and was used to estimate the probability and magnitude of effects to flocks of birds that frequent aldicarb-treated fields. One hundred thirty-five exposure scenarios were modeled that together include a range of bird species, crops, application methods and rates, and regions in the United States. The results indicated that, even for the most sensitive bird species, the risks associated with the agricultural use of granular aldicarb are negligible to low. There are several reasons for the limited risk: 1) the Temik formulation includes a gypsum core and a graphite coating and is black in color, all of which have been shown to be unattractive to birds, and 2) the pesticide is applied subsurface and rapidly dissolves following contact with water. The fact that no bird kill incidents involving appropriate label uses of aldicarb have been conclusively documented in the United States over its 38 years of use supports the results of this refined risk assessment.


Relationship between Dislocation Density and Antibacterial Activity of Cryo-Rolled and Cold-Rolled Copper.
In the present work, cold rolling and cryo-rolling were performed on 99% commercially pure copper substrates. Both cold and cryo-rolling processes caused severe plastic deformation that led to an increase in dislocation density by 14× and 28× respectively, as compared to the pristine material. Increases in average tensile strengths, by 75% (488 MPa) and 150% (698 MPa), were observed in the two rolled materials as the result of the enhancement in dislocation density. In addition to strength, enhanced antibacterial property of cryo-rolled copper was observed in comparison to cold rolled and pristine copper. Initial adhesion and subsequent proliferation of bio-film forming Gram-positive bacteria Staphylococcus aureus was reduced by 66% and 100% respectively for cryo-rolled copper. Approximately 55% protein leakage, as well as ethidium bromide (EtBr) uptake, were observed confirming rupture of cell membrane of S. aureus. Inductively coupled plasma-mass spectroscopy reveals higher leaching of elemental copper in nutrient broth media from the cryo-rolled copper. Detailed investigations showed that increased dislocation led to leaching of copper ions that caused damage to the bacterial cell wall and consequently killing of bacterial cells. Cryo-rolling enhanced both strength, as well as antibacterial activity, due to the presence of dislocations.


[A Successful Case of Conversion Therapy after Chemotherapy for Advanced Rectal Cancer with Inguinal Lymph Node Metastasis].
A70-year-old man was diagnosed with lower rectal cancer with right inguinal lymph node metastasis. Surgical resection was not suitable for this patient. Chemotherapy(IRIS plus bevacizumab followed by XELOX plus bevacizumab)was administered for 16 months. After the chemotherapy, the rectal tumor and lymph node swelling were significantly reduced and distant metastasis was not observed on CT scans. Therefore, the tumor was considered to be resectable and abdominoperineal resection of the rectum with lymph node dissection was performed. On histopathological examination, cancer cells partially remained in the rectal tumor and lateral lymph nodes, although there were no cancer cells in the inguinal lymph nodes. The patient is alive without recurrence a year later. In cases with a good response to chemotherapy, conversion therapy may become an important therapeutic option.


Development of a stroke and neurological rehabilitation unit in a district general hospital.
We describe the experience of setting up a 16-bed unit for the rehabilitation of non-progressive and progressive neurological disorders. Its aim was the active involvement of patients and their families in the rehabilitation process. Various innovations were instigated by the multidisciplinary teams, including early visits home and a leisure service. Over the first two years, 318 patients were admitted to the unit, 51% attributable to stroke.


The effect of chemotherapy on the morphology of human breast carcinoma.
Between biopsy diagnosis and mastectomy 16 patients with carcinoma of the breast received at least three courses of chemotherapy. When the biopsies and the tumour in the mastectomy specimens were compared, an increase in nuclear size, pleomorphism, vacuolation/chromatin clumping and nucleolar prominence were seen in 14 cases. Cytoplasmic changes included accumulation of lipid. There was no correlation between these morphological changes and the disease-free survival period and they could not be used to predict response to chemotherapy.


Lt Paul R. Wolff, MC, USNR, saves crewmen during ordeal within collapsed aircraft fuselage after crash.
Lt Paul R. Wolff, MC, USNR, is the first to receive Pride and Professionalism recognition for his heroic and professional action while responding to the crash of a Japanese Maritime Self Defense Force PS-1 aircraft. At the time of the crash, Dr. Wolff was assigned as the senior flight surgeon representing the First Marine Aircraft Wing and Naval Regional Medical Center Branch Clinic, Marine Corps Air Station, Iwakuni, Japan. On 26 April, 1983, the aircraft crashed just outside the perimeter of the Marine Corps Air Station. Lt Wolff was the first medical person to arrive at the crash site and immediately took charge of the medical aspects of the rescue. Upon learning that several air-crew members were still trapped inside the smoldering wreckage, he unhesitatingly and with complete disregard for his personal safety while fully aware of the personal dangers involved, entered the wreckage to provide medical care to the injured during a 4-hour period. Lt Wolff's bravery, superlative knowledge, exceptional skills, and outstanding leadership abilities under extremely dangerous circumstances led to saving the lives of three crewmen who were trapped. Below is a narration of the events that took place after the crash and Lt Wolff's medical response in the ensuing hours. It is given by Lt Wolff as he recounts the events.


[Melanotic neuroectodermal tumor of infancy].
The melanotic neuroectodermal tumor of infancy is a rare and so far as being classified neoplasm with a high rate of recurrence for one year after diagnosis. Since Krompecher described 1918 the tumor at first, only about 200 cases are reported until today, mostly with manifestation in the maxillary region. The authors present two infants at the age of six and eight weeks with first clinical manifestation of the tumor in the maxillary region. Although there were no other common signs, the tumor destroyed wide areas of the mid-face. In spite of a treatment with radical surgery, recurrences occur rapidly in the first living year. Our clinical and histological findings show characteristics of local malignant growth. For these facts the radical resections of the primary tumor and its recurrences are individually the therapeutical consequences. A follow up of seven years of one infant shows a hypoplasm of the mid-face as a result of the inhibition of further growth by the loss of germs after maxillary hemisection.


Theoretical study on the gas phase reaction of CH2O + NH3: the formation of CH2ONH3, NH2CH2OH, or CH2NH + H2O.
The gas phase reaction between CH2O and NH3 is an important reaction in cold interstellar clouds, combustion chemistry and organic chemistry. In this study, the stationary point on the potential energy surfaces (PESs) for the CH2O + NH3 reaction was computed at the CCSD(T)/6-311++G(3df,3pd)//M06-2X/6-311++G(3df,3pd) level. The temperature- and pressure-dependent rate constants were computed using advanced kinetic models, including microcanonical variational transition state theory and Rice-Ramsperger-Kassel-Marcus (RRKM)/master equation (ME) techniques. Our result predicts that the CH2O + NH3 reaction forms a collisionally thermalized CH2ONH3 complex with respect to thermal unimolecular dissociation and the other products, i.e., NH2CH2OH and CH2NH + H2O, are negligible under atmospheric conditions. The calculated atmospheric lifetime of the CH2ONH3 complex is ∼17 min, which suggests that the CH2ONH3 complex can react with other atmospheric species. The results also suggest that the formation of CH2NH and H2O from the Strecker's process is negligibly small under all the conditions studied here. The decay rate of CH2O + NH3 (5.1 × 10-4 s-1 at 1500 K) suggests that aminomethanol (NH2CH2OH) is likely to occur in the high-temperature combustion of biomass burning, but the rate of formation of NH2CH2OH is negligible under atmospheric conditions. The predicted atmospheric lifetime (∼4 days) of NH2CH2OH in the presence of the OH radical suggests that further reactions with other atmospheric species are possible. The formation of the NH2CHOH radical from the reaction OH + NH2CH2OH can lead to carcinogenic products, such as nitrosamines, acetamide, hydrocycnic acid, NH2 and CO2.


Dental and oral care for chronically ill children and adolescents.
Dental care is the most unmet health care need of children and adolescents who have chronic conditions and diseases. Because advances in therapy and medical technology have extended the lives of this population, it is very likely that these children will seek care in general dental practices. It is extremely important for dental practitioners to be knowledgeable about their patients' history and medical treatment to deliver care safely. This article reviews four chronic diseases of childhood (asthma, cystic fibrosis, cancer, and sickle cell disease) and discusses recommendations for oral and dental treatment.


Studying cytoskeletal dynamics in living cells using green fluorescent protein.
Microfilaments, intermediate filaments, and microtubules are three major cytoskeletal systems providing cells with stability to maintain proper shape. Although the word "cytoskeleton" implicates rigidity, it is quite dynamic exhibiting constant changes within cells. In addition to providing cell stability, it participates in a variety of essential and dynamic cellular processes including cell migration, cell division, intracellular transport, vesicular trafficking, and organelle morphogenesis. During the past eight years since the green fluorescent protein (GFP) was first used as a marker for the exogenous gene expression, it has been an especially booming era for live cell observations of intracellular movement of many proteins. Because of the dynamic behavior of the cytoskeleton in the cell, GFP has naturally been a vital part of the studies of the cytoskeleton and its associated proteins. In this article, we will describe the advantage of using GFP and how it has been used to study cytoskeletal proteins.


Glioma-associated antigens defined by monoclonal antibodies against an avian sarcoma virus-induced rat astrocytoma.
S69-c15 is a highly immunogenic cell line derived from an avian sarcoma virus (ASV)-induced astrocytoma in F-344 rats. Monoclonal antibody (Mab) production was attempted by fusing F-344 rat splenocytes and mouse P3 X 63/Ag8.653 myeloma cells after a syngeneic immunization protocol. 336 fusion clones were screened by cell surface radioimmunoassay (CS-RIA) against the immunizing line S69-c15, rat kidney fibroblast line S203-c11 and Walker rat carcinoma line. Mabs 7G4, 9F1, 10E3 and 10E7 which reacted only with S69-c15 were chosen. Further analysis demonstrated that these Mabs reacted only with rat (13/23 astrocytomas, 2/4 gliomas, 1/11 neurinomas) or mouse (2/10 astrocytomas) neurogenic tumor cells induced by both viral and chemical agents. Reciprocal competition assays suggested that 7G4, 9F1 and 10E3 recognized the same epitope and that 10E7 reacted with a spatially close determinant. Antigen activity could not be found in adult rat tissues (brain, heart, lung, liver, kidney, spleen, thymus, intestine, muscle and peripheral nerve) and fetal brain (8, 12, 20 days gestation) by either absorption analysis or tissue staining. Preliminary characterization indicated that the epitope may be polypeptide-associated. Further antigen purification and tumor localization can be attempted with these Mabs.


Hollow AgI:Ag nanoframes as solar photocatalysts for hydrogen generation from water reduction.
A facile strategy based on the principle of the Kirkendall effect has been developed to synthesize hollow nanoframes and nanoshells of AgI:Ag composites through the controlled anion-exchange reaction between I(-) ions and solid AgBr:Ag (or AgCl:Ag) nanoparticles that serve as templates. Regardless of the morphologies of the template nanoparticles, they can be chemically transformed to hollow AgI:Ag structures with morphologies similar to those of the templates. The synthesized hollow AgI:Ag nanostructures can be used as efficient photocatalysts for H2 generation from water reduction and the decomposition of organic pollutants owing to the enhanced absorption of visible light by the Ag components in the hybrid nanostructures. The hollow nanostructures exhibit a higher photocatalytic performance than the corresponding solid nanoparticles possibly because of the large surface area and unique AgI/Ag interfaces associated with the hollow nanostructures.


Three-dimensional shape analysis using local shape descriptors.
The three-dimensional shape analysis problem is a very demanding test of shape analysis algorithms. Previous approaches to the problem have employed global features such as moments and Fourier descriptors. Global features lack the capacity for solving the partial shape recognition problem, in which only part of the unknown shape is available. Previous approaches to local shape analysis have employed structural (syntactic) methods, but these methods have so far failed to solve the three-dimensional problem. This paper describes a hybrid structural/statistical local shape analysis algorithm which is applied to the three-dimensional problem.


The epidemiological transition in maternal and child health in a Brazilian city, 1982-93: a comparison of two population-based cohorts.
Two studies carried out in 1982 and 1993 in the city of Pelotas, Southern Brazil, provide a unique opportunity for assessing the impact on maternal and child health of the economic and health care changes, which took place in Brazil in this period. The cohorts of mothers and infants of 1982 and 1993 were studied from the time of delivery. In both years, all mothers identified in the city's maternity hospitals answered a standardised questionnaire and their infants were examined. Over 99% of all children born in the city in each of the 2 years were included in the cohorts. Deaths occurring among these children were monitored prospectively, as well as all hospital admissions in the 1993 cohort. In the 1982 study, attempts were made to locate a 25% sample of the children at the mean age of 12 months using the addresses collected at the hospital (82% of the children were located), and all of the cohort children at the mean age of 20 months and 42 months, through a city census (87% were located in both follow-ups). In the 1993 study, 20% of all children plus all low birthweight infants were sought at 12 months of age, using the addresses collected at the hospital, and 95% were successfully traced. There was a 12% fall in the number of births occurring in 1993 (5,304 births), in comparison with 1982 (6,011 births), in spite of the increase in the population of reproductive age in the city during the decade. There was a marked difference in maternal height and weight at the beginning of pregnancy, with women giving birth in 1993 being, on average, 3.4 cm taller and 2.5 kg heavier than those who gave birth in 1982. The proportion of preterm babies (<37 weeks), measured by the date of last menstrual period, increased from 5.6% in 1982 to 7.5% in 1993. The median duration of breast feeding increased from 3.1 months in 1982 to 4.0 months in 1993. At 12 months of age, the prevalence of deficit of weight for age decreased from 5.4% in 1982 to 3.7% in 1993. The prevalence of deficit of height for age, however, increased from 5.3% to 6.1%. The perinatal mortality rate dropped 31%, from 32.2 per 1,000 births in 1982 to 22.1 deaths per 1,000 births in 1993. There was also a marked reduction in the infant mortality rate, from 36.4 per 1,000 livebirths in 1982 to 21.1 per 1,000 livebirths in 1993. The findings of the study indicate that there were improvements in the decade for most of the indicators evaluated, with the exception of birthweight and gestational age. It appears that improvements in perinatal and infant mortality rates are largely due to improvements in the health care sector.


Flavone acetic acid directly induces expression of cytokine genes in mouse splenic leukocytes but not in human peripheral blood leukocytes.
Flavone-8-acetic acid (FAA) is a flavonoid drug that augments mouse natural killer activity, induces cytokine gene expression, and synergizes with recombinant interleukin 2 for the treatment of murine renal cancer. However, FAA has been largely inactive in human clinical trials. In the present study we investigated the ability of FAA treatment to directly induce cytokine mRNA expression in total mouse splenic leukocytes and selected leukocyte subsets, as well as in total human peripheral blood leukocytes. Analysis of RNA isolated from FAA-treated mouse splenic leukocytes demonstrated that treatment with greater than or equal to 100 micrograms/ml of FAA induced expression of tumor necrosis factor alpha (TNF-alpha) mRNA by 1 h and induced maximal expression of TNF-alpha, alpha-interferon, and gamma-interferon mRNA within 3 h. The expression of all cytokine genes was diminished by 6 h. Interferon biological activity was detected in the supernatants of mouse splenic or peripheral blood leukocytes after treatment with FAA. These results correlate well with the previously reported induction of cytokine mRNA genes and biological activity by FAA in vivo. In contrast, FAA did not induce detectable mRNA expression or cytokine protein secretion by human peripheral blood leukocytes under similar conditions. These results demonstrate that FAA can directly stimulate cytokine gene expression in mouse but not in human leukocytes. Further studies performed with highly purified positively selected mouse CD4+ or CD8+ splenic T-lymphocytes, as well as purified B-cells, demonstrated that the FAA-induced expression of gamma-interferon mRNA was mainly induced in the CD8+ lymphocyte subset. alpha-Interferon mRNA was expressed largely in the B-cell population, while TNF-alpha mRNA was induced in all leukocyte subsets tested. Therefore, these results suggest that the immunomodulatory effects of FAA in mice are direct, but different cytokines are induced from different leukocyte subsets. Further, the data suggest that flavonoid compounds or analogues that stimulate cytokine gene expression in human cells might be therapeutically active in cancer patients.


High prevalence of rheumatic heart disease in schoolchildren detected by echocardiography screening in New Caledonia.
Despite the well-documented burden of rheumatic heart disease (RHD) in several Pacific countries, the disease is poorly understood in New Caledonia. The aim of this study was to assess the prevalence of RHD detected by echocardiographic screening in school children. An annual RHD screening programme is conducted by the Health and Social Agency of New Caledonia for school-aged children in their fourth year of primary school. For the purpose of this study, we used data collected during this echocardiographic screening between 2008 and 2010. Of 12,728 children screened, 50.2% were male and the mean age was 9.6 ± 0.6 years. Between 2008 and 2010, 114 children had RHD, corresponding to a prevalence of 8.9 cases per 1000 (95% confidence interval (CI) (7.3-10.6)). Prevalence of RHD was higher on the main island outside Greater Noumea (13.7 per 1000; 95% CI (9.8-17.5)) and in the outlying island groups (14.6 per 1000; 95% CI (8.4-20.9)) than in Greater Noumea (5.8 per 1000; 95% CI (4.1-7.5)). RHD was more prevalent in Melanesian children (13.5 per 1000; 95% CI (10.9-16.1)) than in European (1.8 per 1000; 95% CI (0.4-3.1)). This study documented a high prevalence of RHD in New Caledonia, particularly in districts located outside Noumea and in children of Melanesian heritage. These results uncover a hitherto unknown burden of disease in New Caledonia and underline the importance of delivering secondary prophylaxis to reduce the prevalence of RHD.


Solution structure of the pro-hormone convertase 1 pro-domain from Mus musculus.
The solution structure of the mouse pro-hormone convertase (PC) 1 pro-domain was determined using heteronuclear NMR spectroscopy and is the first structure to be obtained for any of the domains in the convertase family. The ensemble of NMR-derived structures shows a well-ordered core consisting of a four-stranded antiparallel beta-sheet with two alpha-helices packed against one side of this sheet. Sequence homology suggests that the other eukaryotic PC pro-domains will have the same overall fold and most of the residues forming the hydrophobic core of PC1 are highly conserved within the PC family. However, some of the core residues are predicted by homology to be replaced by polar amino acid residues in other PC pro-domains and this may help to explain their marginal stability. Interestingly, the folding topology observed here is also seen for the pro-domain of bacterial subtilisin despite little or no sequence homology. Both the prokaryotic and eukaryotic structures have hydrophobic residues clustered on the solvent-accessible surface of their beta-sheets although the individual residue types differ. In the bacterial case this region is buried at the binding interface with the catalytic domain and, in the eukaryotic PC family, these surface residues are conserved. We therefore propose that the hydrophobic patch in the PC1 pro-domain is involved in the binding interface with its cognate catalytic domain in a similar manner to that seen for the bacterial system. The PC1 pro-domain structure also reveals potential mechanisms for the acid-induced dissociation of the complex between pro- and catalytic domains.


Energy metabolism and protein balance in growing rats fed different levels of dietary fibre and protein.
A study was performed to investigate the effect of different levels of dietary fibre (DF) and dietary protein on visceral organ size, digestibility, nitrogen balance and energy metabolism in rats. Thirty-six male Wistar rats, initial body weight about 76 g were used in a factorial design consisting of three levels of DF (low, 100 g/kg DM: medium, 250 g/kg DM and high, 290 g/kg DM) and two levels of dietary protein (low, 120 g/kg DM and high, 223 g/kg DM). The added fibre source was soybean hulls and Danish fish meal was used as sole source of dietary protein. Measurements of gas-exchange were done on six rats (one group) while urine and faeces were collected individually. The ratio of food/empty body gain increased (P<0.05) with increasing DF and decreasing levels of dietary protein. The weight of the digestive tract was larger (P<0.05) in rats fed the high fibre diet than in those fed the low fibre diet. The digestibility of nutrients and energy decreased linearly with increasing level of soybean fibre (P<0.05). An increased intake of DF was associated with a concomitant loss of protein and energy to faeces. The microbial degradation of NSP and other unabsorbed carbohydrates caused considerably changes in N metabolism of the colon. In rats fed the low protein diets increased levels of DF decreased N excretion in urine and increased N excretion in faeces, while the ratio of retained/digested protein remained constant. When rats were fed the high protein diet protein retention dropped in response to DF both absolute and relative to digested amount, indicating that energy intake could be a limiting factor. Heat production as a percentage of metabolizable energy (HP/ME) was higher (P<0.05) in rats fed the low protein diet than in rats fed the high protein diet, but no significant difference was found among DF levels.