Discrete annular regions of texture contribute independently to the analysis of shape from texture.
Radial frequency (RF) textures (created by applying a sinusoidal modulation of orientation to an otherwise circular texture) have been shown to be globally processed. RF textures differ from RF patterns (paths deformed from circular by a sinusoidal modulation in radius) in that the elements need not be constrained to a specific path. In the natural environment, objects differ from their background in texture, and a bounding contour can mark this textural change. This study examines the extent to which modulation of texture sums across space and whether the inclusion of a boundary between two areas provides a segmentation cue that limits the area over which summation occurs. RF textures were split into two annular regions and signal introduced to inner, outer, or both annuli Thresholds for the detection of RF modulation of orientation were not affected by the presence of a boundary. Further, it was found that the thresholds matched predictions for the independent contribution of the inner and outer areas to performance and that changing the relative phase of the modulation in the inner and outer annuli had no impact on performance, implying independent integration within the two annuli. Finally, integration of modulation information within the annuli was confirmed to ensure these results do apply to textures that are globally processed.


Late kidney allograft loss: what we know about it, and what we can do about it.
Despite dramatic improvements in immunosuppression, late graft loss after kidney transplantation remains a common and difficult problem. Histologic evaluation may reveal changes related to BK polyomavirus infection, hypertension, or calcineurin inhibitor toxicity, which can help to guide therapy. The designation chronic allograft nephropathy should thus be reserved for biopsies with tubular atrophy and interstitial fibrosis without an apparent cause. Although the cause clearly includes both antigen-dependent and antigen-independent events, the approach remains largely to exclude immune mechanisms. Although this review discusses the potential contribution of antibody to chronic injury, it focuses on the basic elements of kidney injury, the role of parenchymal cells in promoting injury, and the proliferative and inflammatory responses that accompanying injury. Strategies to manage these recipients include close attention to accompanying hypertension, diabetes, and hyperlipidemia, as well as consideration for altering immunosuppression; however, therapies that limit epithelial-to-mesenchymal transition or directly block fibrosis pathways may reduce chronic allograft fibrosis and may prove to be useful. Understanding the basic pathogenesis sufficiently to allow early intervention may finally benefit patients who are at high risk for tubular atrophy and interstitial fibrosis and promote their long-term graft function.


Epithelial-derived IL-33 and its receptor ST2 are dysregulated in ulcerative colitis and in experimental Th1/Th2 driven enteritis.
IL-33 is a novel member of the IL-1 family and ligand for the IL-1 receptor-related protein, ST2. Recent evidence suggests that the IL-33/ST2 axis plays a critical role in several autoimmune and inflammatory disorders; however, its role in inflammatory bowel disease (IBD) has not been clearly defined. We characterized IL-33 and ST2 expression and modulation after conventional anti-TNF therapy in Crohn's disease and ulcerative colitis (UC) patients and investigated the role of IL-33 in SAMP1/YitFc (SAMP) mice, a mixed Th1/Th2 model of IBD. Our results showed a specific increase of mucosal IL-33 in active UC, localized primarily to intestinal epithelial cells (IEC) and colonic inflammatory infiltrates. Importantly, increased expression of full-length IL-33, representing the most bioactive form, was detected in UC epithelium, whereas elevated levels of cleaved IL-33 were present in IBD serum. ST2 isoforms were differentially modulated in UC epithelium, and sST2, a soluble decoy receptor with anti-inflammatory properties, was also elevated in IBD serum. Infliximab (anti-TNF) treatment of UC decreased circulating IL-33 and increased sST2, whereas stimulation of HT-29 IEC confirmed IL-33 and sST2 regulation by TNF. Similarly, IL-33 significantly increased and correlated with disease severity, and potently induced IL-5, IL-6, and IL-17 from mucosal immune cells in SAMP mice. Taken together, the IL-33/ST2 system plays an important role in IBD and experimental colitis, is modulated by anti-TNF therapy, and may represent a specific biomarker for active UC.


Lack of evidence of HTLV-I/II infection in T CD8 malignant or reactive lymphoproliferative disorders in France: a serological and/or molecular study of 169 cases.
Human T lymphotropic virus type II (HTLV-II), originally isolated in 1982 from a patient with a "T hairy cell leukemia", has not yet been proven to be the causative agent of any specific hematological disease. In order to screen for such an event, and because HTLV-II has a preferential tropism for OKT8 (CD8) T cells (both in vivo and in vitro), we searched for the presence of HTLV-II in lymphoproliferative diseases (LP) of CD8+ T cells. We report a serological and/or molecular study of 169 patients with a T CD8 LP, including 76 patients with malignant or reactive T CD8 LP (34 lymphomas, 27 large granular leukemias, three prolymphocytic leukemias, one hairy cell leukemia, 11 reactive T CD8 LP) and 93 HIV-1+ patients with a T CD8 peripheral lymphocytosis ( > 1500/mm3) from a prospective HIV cohort involving 1264 individuals. In the first series, the 40 sera available were all HTLV-I/II negative, except a 67-year-old French Guyanan man, with a cutaneous large T CD8 cell lymphoma, HTLV-I+. Furthermore, the molecular analysis of the 69 available DNA samples by PCR failed to detect any proviral HTLV-I/II sequences, except for the HTLV-I+ patient. The serological study of the 93 HIV-1+ individuals with CD8 lymphocytosis, showed that three patients were HTLV-I+, but none was HTLV-II+. Thus, in contrast to HTLV-I, whose etiological role in adult T cell leukemia is now well established, there is neither epidemiological nor molecular evidence that prototypic HTLV-II may be etiologically associated specifically with any of the CD8+ T cell LP investigated in this report.


Anticancer Effects of Wild Mountain Mentha longifolia Extract in Adrenocortical Tumor Cell Models.
Mint [Mentha longifolia (L.) Hudson] is an aromatic plant that belongs to Lamiaceae family. It is traditionally used as herbal tea in Europe, Australia and North Africa and shows numerous pharmacological effects, such as spasmolytic, antioxidant, antimicrobial and anti-hemolytic. Recently, its antiproliferative role has been suggested in a small number of tumor cell models, but no data are available on adrenocortical carcinoma, a malignancy with a survival rate at 5 years of 20%-30% which frequently metastasize. This work aimed to study the effects of Mentha longifolia L. crude extract (ME) on two adrenocortical tumor cell models (H295R and SW13 cells). Chemical composition of ME was assessed by gas-chromatography/mass spectrometry and NMR spectroscopy analysis. Brine shrimp lethality assay showed ME effects at >0.5 µg/µl (p < 0.05). Cell viability and vitality were determined by MTT, SRB, and trypan blue assays in H295R and SW13 cells. The anti-proliferative effects of ME were more evident in SW13 cells at 72 h (ME > 0.5 µg/µl, p < 0.05). Combination of ME with mitotane (approved drug for adrenocortical carcinoma) seemed not to reinforce the efficacy of the herb. As control, human fibroblasts were treated with ME with no effect on cell viability. Clonogenic assay was concordant with previous cell viability tests (ME > 0.5 µg/µl, p < 0.05), while Wright staining demonstrated the presence of both necrotic and apoptotic cells. Cell cycle analysis showed a strong increase in subG0/G1 phase, related to cell death. Furthermore, MAPK and PI3k/Akt pathways were modulated by Western blot analysis when treating cells with ME alone or combined with mitotane. The crude methanolic extract of wild mountain mint can decrease cell viability, vitality and survival of adrenocortical tumor cell models, in particular of SW13 cells. These data show the potential anticancer effects of ME, still more work is needed to corroborate these findings.


High rates of clinical and subclinical tuberculosis among HIV-infected ambulatory subjects in Tanzania.
We sought to determine the prevalence of active tuberculosis among ambulatory HIV-infected persons in Tanzania with CD4 cell counts of > or =200 cells/mm3 and a bacille Calmette-Guerin vaccination scar. Subjects who volunteered for a tuberculosis booster vaccine trial were screened for active tuberculosis by obtainment of a history, physical examination, chest radiography, sputum culture and acid fast bacillus (AFB) stain, and blood culture. All subjects underwent a tuberculin skin test (TST) and lymphocyte proliferation assays (LPAs) for detection of responses to mycobacterial antigens. Active tuberculosis was identified at baseline in 14 (15%) of the first 93 subjects who were enrolled: 10 (71%) had clinical tuberculosis (symptoms or chest radiograph findings), and 4 (29%) had subclinical tuberculosis (positive sputum AFB stain or culture results but no symptoms or chest radiograph findings). An additional 6 subjects with subclinical tuberculosis were identified subsequently. The 10 subjects with subclinical tuberculosis included 3 with positive sputum AFB stains results and 7 who were only identified by a positive sputum culture result. Compared with subjects who did not have tuberculosis, the 10 subjects with subclinical tuberculosis were more likely to have peripheral lymphadenopathy, positive TST results, and elevated LPA responses to early secreted antigenic target-6 (ESAT). Eight of 10 patients had received isoniazid because of a positive TST result before active tuberculosis was recognized. Clinical and subclinical tuberculosis are common among ambulatory HIV-infected persons, and some cases can only be identified by sputum culture. World Health Organization guidelines for screening for latent tuberculosis before treatment do not recommend sputum culture and, therefore, may fail to identify a substantial number of HIV-infected persons with subclinical, active tuberculosis.


Fatal hypermagnesemia caused by an Epsom salt enema: a case illustration.
The authors describe a case of fatal hypermagnesemia caused by an Epsom salt enema. A 7-year-old male presented with cardiac arrest and was found to have a serum magnesium level of 41.2 mg/dL (33.9 mEq/L) after having received an Epsom salt enema earlier that day. The medical history of Epsom salt, the common causes and symptoms of hypermagnesemia, and the treatment of hypermagnesemia are reviewed. The easy availability of magnesium, the subtle initial symptoms of hypermagnesemia, and the need for education about the toxicity of magnesium should be of interest to physicians.


Changes in luminal pH caused by calcium release in sarcoplasmic reticulum vesicles.
Fast (milliseconds) Ca2+ release from sarcoplasmic reticulum is an essential step in muscle contraction. To electrically compensate the charge deficit generated by calcium release, concomitant fluxes of other ions are required. In this study we investigated the possible participation of protons as counterions during calcium release. Triad-enriched sarcoplasmic reticulum vesicles, isolated from rabbit fast skeletal muscle, were passively loaded with 1 mM CaCl2 and release was induced at pCa = 5.0 and pH = 7.0 in a stopped-flow fluorimeter. Accompanying changes in vesicular lumen pH were measured with a trapped fluorescent pH indicator (pyranin). Significant acidification (approximately 0.2 pH units) of the lumen occurred within the same time scale (t(1/2) = 0.75 s) as calcium release. Enhancing calcium release with ATP or the ATP analog 5'-adenylylimidodiphosphate (AMPPNP) produced >20-fold faster acidification rates. In contrast, when calcium release induced with calcium with or without AMPPNP was blocked by Mg2+, no acidification of the lumen was observed. In all cases, rate constants of luminal acidification corresponded with reported values of calcium release rate constants. We conclude that proton fluxes account for part (5-10%) of the necessary charge compensation during calcium release. The possible relevance of these findings to the physiology of muscle cells is discussed.


Accuracy of aortic annular measurements obtained from three-dimensional echocardiography, CT and MRI: human in vitro and in vivo studies.
To determine the accuracy of calcium-containing rings measurements imaged by three-dimensional echocardiography (3DE), multi-slice CT (MSCT) and cardiac magnetic resonance (CMR) under ideal conditions against the true ring dimensions. To compare the accuracy of aortic annulus (AoA) measurements in ex vivo human hearts using 3DE, MSCT and CMR. To determine the accuracy of AoA measurements in an in vivo human model. 3DE, MSCT and CMR imaging were performed on 30 calcium-containing rings and 28 explanted human hearts. Additionally, 15 human subjects with clinical indication for MSCT underwent 3DE. Two experts in each modality measured the images. Bias and intraclass correlation coefficient for accuracy of imaging measurements when compared with actual ring dimensions. Bias, intraclass correlation coefficient and variability were obtained: (1) when comparing explanted human heart AoA measurements from the two remaining imaging modalities with the most accurate one as determined from the ring measurements and (2) in in vivo human AoA measurements. Analysis was repeated on explanted heart subgroups divided by aortic valve Agatston score. Against the known ring dimensions, CMR had the highest accuracy and the lowest variability. MSCT measurements had high accuracy but wider variability and 3DE had the lowest accuracy with the largest variability. When 3DE and MSCT were compared with CMR, 3DE underestimated and MSCT overestimated AoA dimensions, but inter-measurement variability of 3DE and MSCT were similar. When divided by Agatston score, both 3DE and MSCT measurements were larger and showed greater variability with increasing calcium burden. The in vivo study showed that the correlation between 3DE and MSCT measurements was high; however, 3DE measurements were smaller than those measured with MSCT. In the in vitro model, CMR measurements were the most accurate for assessing the actual dimensions suggesting that further investigations on its role in AoA measurement in TAVR are needed. However from the in vivo model, MSCT and 3DE are reasonable alternatives with the understanding that they can slightly overestimate and underestimate annular dimensions, respectively.


Age at onset and pattern of neuropsychological impairment in mild early-stage Alzheimer disease. A study of a community-based population.
To examine the effects of age at onset on neuropsychological functioning in a group of patients with probable Alzheimer disease (AD) and, within this group, to scrutinize further those patients with mild early-onset disease as it was hypothesized that within this group specific patterns of cognitive impairment could be identified that correlated with neuropathological staging of the disease. Each patient underwent an extensive neuropsychological test battery to examine a wide range of cognitive processes to provide information to identify subtypes of dementia. The Memory Clinic in the Department of Geriatric Medicine, Concord Hospital, Concord, New South Wales, Australia. One hundred forty-five community-residing case patients with probable AD were studied; within this group, 51 case patients with mild AD and a Mini-Mental State Examination score greater than 19 were further examined; 36 similarly aged control patients who were part of a larger case-control study of AD in an urban population were also examined. A diagnosis of probable and possible AD was made if the case patient had evidence of memory impairment and met criteria according to the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association. Individual neuropsychological test scores were compared. The tests were then grouped into 7 cognitive domains. Patterns of early cognitive impairment were derived from these comparisons. With an earlier age at onset, significantly more impairment on tests of digit span and praxis was seen, while the duration of disease had no independent effect once the age at onset was fixed. Patients with mild early-onset dementia and a Mini-Mental State Examination score greater than 19 showed significant impairment in tests of attention, memory, frontal/executive functions, visuospatial ability, praxis, and visual agnosia compared with that shown by control patients. In this group, further analyses revealed that impairment in memory and frontal/ executive functions were the earliest signs of cognitive impairment. These data showed that when the duration of disease was adjusted for, case patients with an earlier age at onset of AD demonstrated significantly more impairment on tests of attention span and working memory (digit span), graphomotor function (copy loops), and apraxia than those with an older age at onset. Our findings support the view that the hippocampus and its connections are affected in the early stages of AD. The deficits in the frontal/executive functions also suggest that a disruption of cortical pathways to the frontal lobes and the pathological changes in this region occur early in the disease.


Autoreactive T Cells and Chronic Fungal Infection Drive Esophageal Carcinogenesis.
Humans with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), a T cell-driven autoimmune disease caused by impaired central tolerance, are susceptible to chronic fungal infection and esophageal squamous cell carcinoma (ESCC). However, the relationship between autoreactive T cells and chronic fungal infection in ESCC development remains unclear. We find that kinase-dead Ikkα knockin mice develop APECED-like phenotypes, including impaired central tolerance, autoreactive T cells, chronic fungal infection, and ESCCs expressing specific human ESCC markers. Using this model, we investigated the link between ESCC and fungal infection. Autoreactive CD4 T cells permit fungal infection and incite tissue injury and inflammation. Antifungal treatment or autoreactive CD4 T cell depletion rescues, whereas oral fungal administration promotes, ESCC development. Inhibition of inflammation or epidermal growth factor receptor (EGFR) activity decreases fungal burden. Fungal infection is highly associated with ESCCs in non-autoimmune human patients. Therefore, autoreactive T cells and chronic fungal infection, fostered by inflammation and epithelial injury, promote ESCC development.


CK5/6 in effusions: no difference between mesothelioma and pulmonary and nonpulmonary adenocarcinoma.
To test the performance of CK5/6 for the differentiation between mesothelioma, adenocarcinoma and benign mesothelia/proliferations in effusion cytology. CKS/6 immunocytochemistry was applied to ethanol-fixed cytospin preparations from 74 benign and malignant effusions. Reactivity was seen in 7 of 8 mesotheliomas and in 9 of 11 benign mesothelial proliferations but also in 11 of l7 pulmonary adenocarcinomas and in 12 of 31 adenocarcinomas of nonpulmonary origin. Reactivity was also found in 3 of 5 non-small cell lung carcinomas and 1 of 1 squamous carcinoma. CK5/6 reactivity was found in a considerable proportion of metastatic adenocarcinomas of pulmonary and nonpulmonary origin. The high reactivity rate in pulmonary adenocarcinomas disagrees with the results obtained with histologic sections from solid tumor tissue, and CK5/6 seems to be of very limited value as an additional marker in effusion cytology.


Haploid genetic screens in human cells identify host factors used by pathogens.
Loss-of-function genetic screens in model organisms have elucidated numerous biological processes, but the diploid genome of mammalian cells has precluded large-scale gene disruption. We used insertional mutagenesis to develop a screening method to generate null alleles in a human cell line haploid for all chromosomes except chromosome 8. Using this approach, we identified host factors essential for infection with influenza and genes encoding important elements of the biosynthetic pathway of diphthamide, which are required for the cytotoxic effects of diphtheria toxin and exotoxin A. We also identified genes needed for the action of cytolethal distending toxin, including a cell-surface protein that interacts with the toxin. This approach has both conceptual and practical parallels with genetic approaches in haploid yeast.


[Prognosis of complex ventricular arrhythmias at different periods in myocardial infarct].
The incidence and pattern of ventricular arrhythmias in acute, subacute, late hospital and posthospital periods of myocardial infarction were investigated, using Holter monitoring, in 121 patients with large-focal and transmural myocardial infarction. A high incidence of complex ventricular arrhythmias was noted after the transfer from intensive care unit (38.8%). The relationship of complex ventricular arrhythmias to ventricular arrhythmias at previous stages was reviewed. Factors contributing to arrhythmias at different stages of myocardial infarction were analysed; the informative value of clinical signs and electric myocardial instability parameters was assessed. Signs having the greatest predictive power were identified, and risk indices for the development of arrhythmias during the subacute and late hospital stages of myocardial infarction were established using linear discriminant analysis, to enable identification of patients at risk of developing ventricular arrhythmias.


Nutrient and dissolved organic carbon removal from water using mining and metallurgical by-products.
Excess nutrient input to water bodies frequently results in algal blooms and development of oxygen deficient conditions. Mining or metallurgical by-products can potentially be utilised as filtration media within water treatment systems such as constructed wetlands, permeable reactive barriers, or drain liners. These materials may offer a cost-effective solution for the removal of nutrients and dissolved organic carbon (DOC) from natural waters. This study investigated steel-making, alumina refining (red mud and red sand) and heavy mineral processing by-products, as well as the low-cost mineral-based material calcined magnesia, in laboratory column trials. Influent water and column effluents were analysed for pH and flow rate, alkalinity, nutrient species and DOC, and a range of major cations and anions. In general, by-products with high Ca or Mg, and to a lesser extent those with high Fe content, were well-suited to nutrient and DOC removal from water. Of the individual materials examined, the heavy mineral processing residue neutralised used acid (NUA) exhibited the highest sorption capacity for P, and removed the greatest proportions of all N species and DOC from influent water. In general, NUA and mixtures containing NUA, particularly those with calcined magnesia or red mud/red sand were the most effective in removing nutrients and DOC from influent water. Post-treatment effluents from columns containing NUA and NUA/steel-making by-product, NUA/red sand and NUA/calcined magnesia mixtures exhibited large reductions in DOC, P and N concentrations and exhibited a shift in nutrient ratios away from potential N- and Si-limitation and towards potential P-limitation. If employed as part of a large-scale water treatment scheme, use of these mining and metallurgical by-products for nutrient removal could result in reduced algal biomass and improved water quality. Identification and effective implementation of mining by-products or blends thereof in constructed wetlands or other intervention structures to augment nutrient and DOC retention has considerable potential as an aquatic ecosystem management tool.


Three-dimensional transesophageal echocardiography in degenerative mitral regurgitation.
The morphology of mitral valve (MV) prolapse and flail may be extremely variable, with dominant and secondary dynamic lesions. Any pathologic valve appears unique and different from any other. Three-dimensional (3D) transesophageal echocardiography is a powerful tool to evaluate the geometry, dynamics, and function of the MV apparatus and may be of enormous value in helping surgeons perform valve repair procedures. Indeed, in contrast to the surgical view, 3D transesophageal echocardiography can visualize MV prolapse and flail in motion and from different perspectives. The purpose of this special article is not to provide a comprehensive review of degenerative MV disease but rather to illustrate different types of mitral prolapse and flail as they appear from multiple 3D transesophageal echocardiographic perspectives using a series of clinical scenarios. Because in everyday practice, 3D transesophageal echocardiographic images of MV prolapse and flail are usually observed in motion, each scenario is accompanied by several videos. Finally, the authors provide for each scenario a brief description of the surgical techniques that are usually performed at their institution.


Consumer education needed on norovirus prevention and control: findings from a nationally representative survey of U.S. adults.
Noroviruses (NoVs) are the leading cause of foodborne disease in the United States; however, little is known about consumers' knowledge of NoV infection and their understanding of how to prevent and control associated illness. A nationally representative Web-enabled panel survey of U.S. adults (n = 1,051) was conducted to collect information on consumers' awareness and knowledge of NoVs. Respondents who had heard of NoVs were asked 22 true-and-false questions on the transmission, prevention, and control of NoVs. Forty-seven percent of respondents reported awareness of NoVs, and 85% of respondents had heard of the terms "cruise ship virus," "the stomach bug," or "the stomach flu," which are commonly used to describe NoVs. Of those respondents who had previously heard of NoV or other terms used by consumers to describe NoV (n = 948), 36% correctly answered 11 or more of the 22 true-and-false questions, suggesting that consumers have limited knowledge on how to prevent and control NoV infection. Most consumers do not understand that the primary mode of transmission for NoV infection is fecal to oral, and many have the misperception that meat and poultry are sources of NoV infection. There is the need to educate consumers about how to prevent and control NoV infection. Although there is a proliferation of food safety education materials available, most focus on foodborne bacteria rather than viruses. The survey results will be used to revise existing consumer food safety educational materials to include information on NoV prevention and control.


Patient, physician & society: Northwestern University Medical School.
Northwestern University Medical School's Patient, Physician & Society (PPS) course was introduced in 1993 as part of a complete restructuring of the first- and second-year curriculum. The PPS course meets two afternoons per week throughout the first two years, with one afternoon focusing on the relationship between patients and physicians and the other on that between physicians and society. The course is designed to provide a comprehensive, integrated introduction to professional skills and perspectives. Fourteen distinct curricular units address personal and professional ethics, medical humanities, behavioral sciences, physician-patient communication, physical diagnosis and clinical reasoning, health services organization and financing, preventive medicine, and the health of vulnerable groups. Health promotion as a primary goal of medicine is an underlying theme throughout the course. Active and interactive learning formats afford many opportunities for personal reflection and discussion. The overall response to the course has been positive, and survey data indicate that students completing PPS report more progress toward the school's fundamental educational goals than do students who had progressed through the first two years before the new curriculum was introduced. Still, a number of students are clearly uncomfortable with educational strategies that give them responsibility for finding answers on their own. Contrasts between PPS and the basic science courses--in content, presentation, and evaluation--highlight the importance of coordinating and integrating the overall medical school curriculum. Plans for enhancing the course include focusing on faculty development and student evaluation, as well as explicitly extending PPS material into the clerkship years.


Assessment of prokaryote to eukaryote ratios in environmental samples by SSU rDNA length polymorphism.
Microbial communities are important regulators of many processes in all ecosystems. Understanding of ecosystem processes requires at least an overview of the involved microorganisms. While in-depth identification of microbial species in environmental samples can be achieved by next generation sequencing, profiling of whole microbial communities can be accomplished via less labour-intensive approaches. Especially automated ribosomal intergenic spacer analysis (ARISA) are of interest as they are highly specific even at fine scales and widely applicable for environmental samples. Yet, established protocols lack the possibility to compare prokaryotic and eukaryotic communities as different primer sets are necessary. However, shifts in the eukaryote to prokaryote ratio can be a useful indicator for ecosystem processes like decomposition or nutrient cycling. We propose a protocol to analyse prokaryotic and eukaryotic communities using a single primer pair based reaction based on a region with variable length (V4, which is about 180 bp shorter in prokaryotes compared to eukaryotes) in the small ribosomal subunit flanked by two highly conservative regions. Shifts in the prokaryotic and eukaryotic ratio between samples can be reliably detected by fragment length polymorphism analysis as well as sequencing of this region. Together with established approaches such as ARISA or 16S and ITS rDNA sequencing, this can provide a more complex insight into microbial community shifts and ecosystem processes.


Modeling the Ecological Niche of Bacillus anthracis to Map Anthrax Risk in Kyrgyzstan.
Anthrax, caused by the environmental bacterium Bacillus anthracis, is an important zoonosis nearly worldwide. In Central Asia, anthrax represents a major veterinary and public health concern. In the Republic of Kyrgyzstan, ongoing anthrax outbreaks have been reported in humans associated with handling infected livestock and contaminated animal by-products such as meat or hides. The current anthrax situation has prompted calls for improved insights into the epidemiology, ecology, and spatial distribution of the disease in Kyrgyzstan to better inform control and surveillance. Disease control for both humans and livestock relies on annual livestock vaccination ahead of outbreaks. Toward this, we used a historic database of livestock anthrax reported from 1932 to 2006 mapped at high resolution to develop an ecological niche model-based prediction of B. anthracis across Kyrgyzstan and identified spatial clusters of livestock anthrax using a cluster morphology statistic. We also defined the seasonality of outbreaks in livestock. Cattle were the most frequently reported across the time period, with the greatest number of cases in late summer months. Our niche models defined four areas as suitable to support pathogen persistence, the plateaus near Talas and Bishkek, the valleys of western Kyrgyzstan along the Fergana Valley, and the low-lying areas along the shore of Lake Isyk-Kul. These areas should be considered "at risk" for livestock anthrax and subsequent human cases. Areas defined by the niche models can be used to prioritize anthrax surveillance and inform efforts to target livestock vaccination campaigns.


Increased Neuroprotective Microglia and Photoreceptor Survival in the Retina from a Peptide Inhibitor of Myeloid Differentiation Factor 88 (MyD88).
Myeloid differentiation factor 88 (MyD88) is an adaptor protein for the Toll-like receptor (TLR) and interleukin 1 receptor (IL-1R) families of innate immunity receptors that mediate inflammatory responses to cellular injury. TLR/IL1R/MyD88 signaling is known to contribute to retinal degeneration, although how MyD88 regulates neuronal survival, and the effect of MyD88 on the inflammatory environment in the retina, is mostly unknown. In this study, we tested the hypothesis that blocking MyD88-mediated signaling early in retinal degeneration promotes transition of microglia towards a neuroprotective anti-inflammatory phenotype, resulting in enhanced photoreceptor survival. We also tested whether systemic delivery of a pharmacologic MyD88 inhibitor has therapeutic potential. The rd10 mouse model of retinal degeneration was injected intraperitoneally with increasing doses of a MyD88 blocking peptide or control peptide early in degeneration, and inflammatory responses and photoreceptor survival were measured at specific time points using flow cytometry, cytokine profiling, and electroretinograms. Our results demonstrated that rd10 mice injected with a low dose of MyD88 inhibitor peptide showed increased rod photoreceptor function and reduced apoptosis compared with control peptide and uninjected mice. MyD88 inhibition also resulted in fewer microglia/macrophage cells in the photoreceptor layer whereas total peripheral and retinal macrophage were not changed. Furthermore, increased number of cells expressing the Arg1 marker of neuroprotective microglia in the photoreceptor layer and higher MCP-1 and anti-inflammatory cytokine IL-27 were associated with photoreceptor survival. Therefore, these data suggest that the MyD88 inhibitor modified the retina environment to become less inflammatory, leading to improved photoreceptor function and survival.


[Posterior lumbar interbody fusion].
This paper reports the posterior lumbar interbody fusion (PLIF) that has been performed on fourteen patients. It includes 6 cases of spondylolytic spondylolisthesis, 3 cases of degenerative spondylolisthesis, 2 cases of postoperative recurrence of lumbar disc protrusion, 2 cases of unstable prolapse of intervertebral disc, 1 cases of consequent spinal canal stenosis after lumbar lamina fusion. As a result 92 per cent of the operations are successful. The method of operation and it's modification are reported in detail. The indication of operation and the evaluative criteria of interbody bone union discussed. The intact of lumbar posterior structures, the condition of bone grating bed, the quality and disperse of bone graft are main factors that influencing bone union. Bleeding from venous plexus of spinal canal and from vertebral cancellous bone, injury of lumbosacral nerve root and cauda equina are the main surgical complications that should be stressed.


Physiology and pathophysiology of eryptosis.
Suicidal erythrocyte death (eryptosis) is characterized by cell shrinkage, cell membrane blebbing, and cell membrane phospholipid scrambling with phosphatidylserine exposure at the cell surface. Eryptotic cells adhere to the vascular wall and are rapidly cleared from circulating blood. Eryptosis is stimulated by an increase in cytosolic Ca(2)+ activity, ceramide, hyperosmotic shock, oxidative stress, energy depletion, hyperthermia, and a wide variety of xenobiotics and endogenous substances. Inhibitors of eryptosis include erythropoietin and nitric oxide. Enhanced eryptosis is observed in diabetes, renal insufficiency, hemolytic uremic syndrome, sepsis, mycoplasma infection, malaria, iron deficiency, sickle cell anemia, beta-thalassemia, glucose-6-phosphate dehydrogenase-(G6PD) deficiency, hereditary spherocytosis, paroxysmal nocturnal hemoglobinuria, Wilson's disease, myelodysplastic syndrome, and phosphate depletion. Eryptosis is further enhanced in gene-targeted mice with deficient annexin 7, cGMP-dependent protein kinase type I (cGKI), AMP-activated protein kinase (AMPK), anion exchanger 1 (AE1), adenomatous polyposis coli (APC), and Klotho, as well as in mouse models of sickle cell anemia and thalassemia. Decreased eryptosis is observed in mice with deficient phosphoinositide-dependent kinase 1 (PDK1), platelet activating factor (PAF) receptor, transient receptor potential channel 6 (TRPC6), janus kinase 3 (JAK3), and taurine transporter (TAUT). Eryptosis may be a useful mechanism to remove defective erythrocytes prior to hemolysis. Excessive eryptosis may, however, compromise microcirculation and lead to anemia.


[Piracetam in anesthesia for prevention of postoperative delirium].
Delirium is a serious postoperative complication after general anaesthesia. The incidence is estimated to be about 10-15%. In the case of additional risk factors, such as old age, previously existing cerebral-vasculous and psychic deficiencies, and anaesthesia of long duration the incidence is much higher. Delirium impairs postoperative mobilisation and convalescence of the patient, can lead to a longer hospital stay and is associated with higher mortality rates. In a series of clinical studies it could be shown that the perioperative administration of the nootropic piracetam led to a shorter recovery period after anaesthesia and had a favourable influence on the delirious symptoms. Especially patients with risk factors for postoperative delirium profited from prophylactic application of piracetam. The success of this medication can be explained by a protective influence of the substance on central neurons against hypoxia, ischemia and intoxication, all of which are discussed as possible causes for postoperative delirium. Next to the pathogenetic mechanisms of postoperative delirium, the mode of action of piracetam is shown in a review and a summary of references on the clinical use in anaesthesia is given.


Histones in Xenopus laevis' early development: the race against time.
Using cloned histone gene variants, the expression of three types of histone gene clusters was studied at the RNA level during early development in Xenopus laevis. For each histone class the number of mRNA molecules per embryo strongly decreased upon ovulation and steadily increased during early development, with a slight decrease at the neurula stage. Variation of the stringency of hybridization revealed that none of the histone genes probed is specifically and uniquely expressed at any time point in embryogenesis. The observed variation of histone mRNA content with time after fertilization is consistent with what is known about rate constants for RNA synthesis and degradation and about histone mRNA storage in the oocyte, provided that approximately 11 h after fertilization a regulatory transition is proposed (KOSTER, DESTREE and WESTERHOFF (1988) J. Theor. Biol. 135, 139-167). Similarly, the observed amounts of histone mRNA could well be sufficient to direct the synthesis of the required amount of histone protein. Control Analysis reveals that late in development, the histone to DNA ratio is controlled virtually equally strongly by the rate constants of DNA replication, transcription and translation. The control coefficient for RNA degradation is only a little smaller, whereas that for proteolysis is negligible. Indeed, the deceleration of DNA synthesis around the Mid Blastula Transition (some 8 h after fertilization in our studies) is a regulatory step that is essential in order to allow Xenopus to synthesize sufficient histones to structure its DNA; otherwise the embryo would run out of histone protein and histone mRNA soon thereafter. A model that assumes that the lengthening of the cell cycle around the Mid Blastula Transition it itself a response to the decrease in the concentration of histone protein not complexed to DNA, is shown to account for the dynamics of histone and DNA synthesis during the first 50 h after fertilization.


Low birth weight is associated with increased sympathetic activity: dependence on genetic factors.
Low birth weight may be associated with high blood pressure in later life through genetic factors, an association that may be explained by alterations in sympathetic and parasympathetic activity. We examined the association of birth weight with cardiac pre-ejection period and respiratory sinus arrhythmia (indicators of cardiac sympathetic and parasympathetic activity, respectively) and with blood pressure in 53 dizygotic and 61 monozygotic adolescent twin pairs. Birth weight of the twins was obtained from the mothers. Pre-ejection period and respiratory sinus arrhythmia were measured with electrocardiography and impedance cardiography at rest, during a reaction time task, and during a mental arithmetic task. In the overall sample, lower birth weight was significantly associated with shorter pre-ejection period at rest, during the reaction time task, and during the mental arithmetic task (P=0.0001, P<0.0001, and P=0.0001, respectively) and with larger pre-ejection period reactivity to the stress tasks (P=0.02 and P=0.06, respectively). In within-pair analyses, differences in birth weight were associated with differences in pre-ejection period at rest and during both stress tasks in dizygotic twin pairs (P=0.01, P=0.06, and P=0.2, respectively) but not in monozygotic twin pairs (P=0.9, P=1.0, and P=0.5, respectively). Shorter pre-ejection period explained approximately 63% to 84% of the birth weight and blood pressure relation. Low birth weight is associated with increased sympathetic activity, and this explains a large part of the association between birth weight and blood pressure. In addition, our findings suggest that the association between birth weight and sympathetic activity depends on genetic factors.


The effects of prostaglandin E-1 and F-2 alpha on maturation of mouse oocytes in vitro.
The direct effects of prostaglandin E-1 (PGE-1) and F-2 alpha (PGF-2alpha) on isolated mouse oocytes were examined in a microtube culture method. In oocytes exposed to PGE-1, meiotic resumption was suppressed at the anaphase I stage, presenting an irregular arrangement of the chromosomes along with the spindle fibres which were characterized by nondisjunction of the tetrads. PGF-2alpha was less potent in suppressing oocyte maturation than PGE-1. The critical time of the action of PGE-1 was between 7-5 and 12 hr of incubation.


Transfecting protoplasts by electroporation to study viroid replication.
Protoplasts have been extensively used to study replication of plant viruses at the cellular level. However, their use in studying viroid replication has been very limited because of various technical barriers. As a result, mechanisms of viroid replication remain poorly understood. The electroporation protocol presented in this unit offers a simple, fast and reproducible method to inoculate protoplasts derived from cultured cells of tobacco (Nicotiana tabacum) and the related species Nicotiana benthamiana to study viroid replication. The protocol has proven to be useful in characterizing sequence/structural features of potato spindle tuber viroid (PSTVd) that are important for replication at the cellular level, and may be adapted to study the replication of other viroids.


Temperamental emotionality in preschool-aged children and depressive disorders in parents: associations in a large community sample.
Researchers and clinicians have long hypothesized that there are temperamental vulnerabilities to depressive disorders. Despite the fact that individual differences in temperament should be evident in early childhood, most studies have focused on older youth and adults. We hypothesized that if early childhood temperament is a risk factor for depressive disorders, it should be associated with better established risk markers, such parental depression. Hence, we examined the associations of laboratory-assessed positive emotionality (PE), negative emotionality (NE), and behavioral inhibition (BI) with semistructured interview-based diagnoses of parental depressive disorders in a community sample of 536 3-year old children. Children with higher levels of NE and BI had higher probabilities of having a depressed parent. However, both main effects were qualified by interactions with child PE. At high and moderate (but not low) levels of child PE, greater NE and BI were associated with higher rates of parental depression. Conversely, at low (but not high and moderate) levels of child NE, low PE was associated with higher rates of parental depression. Child temperament was not associated with parental anxiety and substance use disorders. These findings indicate that laboratory-assessed temperament in young children is associated with parental depressive disorders; however, the relations are complex, and it is important to consider interactions between temperament dimensions rather than focusing exclusively on main effects.


Sleep loss and structural plasticity.
Wakefulness and sleep are dynamic states during which brain functioning is modified and shaped. Sleep loss is detrimental to many brain functions and results in structural changes localized at synapses in the nervous system. In this review, we present and discuss some of the latest observations of structural changes following sleep loss in some vertebrates and insects. We also emphasize that these changes are region-specific and cell type-specific and that, most importantly, these structural modifications have functional roles in sleep regulation and brain functions. Selected mechanisms driving structural modifications occurring with sleep loss are also discussed. Overall, recent research highlights that extending wakefulness impacts synapse number and shape, which in turn regulate sleep need and sleep-dependent learning/memory.


Impact of body mass index on growth hormone stimulation tests in children and adolescents: a systematic review and meta-analysis.
Peak stimulated growth hormone (GH) levels are known to decrease with increasing body mass index (BMI), possibly leading to overdiagnosis of GH deficiency (GHD) in children with overweight and obesity. However, current guidelines do not guide how to interpret the peak GH values of these children. This systematic review and meta-analysis aimed to study the effect of the BMI standard deviation score (SDS) on stimulated peak GH values in children, to identify potential moderators of this association, and to quantify the extent to which peak GH values in children with obesity are decreased. This systematic review was performed by the PRISMA guidelines. Medline, Embase, Cochrane, Web of Science, and Google Scholar databases were searched for studies reporting the impact of weight status on peak GH in children. Where possible, individual participant data was extracted and/or obtained from authors. Quality and risk of bias were evaluated using the Scottish Intercollegiate Guidelines Network (SIGN) checklists. The primary outcome was the association between peak GH values and BMI SDS. The pooled correlation coefficient r, 95% confidence interval (CI), and heterogeneity statistic I2 were calculated under a multilevel, random-effects model. In addition, exploratory moderator analyses and meta-regressions were performed to investigate the effects of sex, pubertal status, presence of syndromic obesity, mean age and mean BMI SDS on the study level. For the individual participant dataset, linear mixed-models regression analysis was performed with BMI SDS as the predictor and ln(peak GH) as the outcome, accounting for the different studies and GH stimulation agents used. In total, 58 studies were included, providing data on n = 5135 children (576 with individual participant data). Thirty-six (62%) studies had high, 19 (33%) medium, and 3 (5%) low risks of bias. Across all studies, a pooled r of -0.32 (95% CI -0.41 to -0.23, n = 2434 patients from k = 29 subcohorts, I2 = 75.2%) was found. In meta-regressions, larger proportions of males included were associated with weaker negative correlations (p = 0.04). Pubertal status, presence of syndromic obesity, mean age, and mean BMI SDS did not moderate the pooled r (all p > 0.05). Individual participant data analysis revealed a beta of -0.123 (95% CI -0.160 to -0.086, p < 0.0001), i.e. per one-point increase in BMI SDS, peak GH decreases by 11.6% (95% CI 8.3-14.8%). To our knowledge, this is the first systematic review and meta-analysis to investigate the impact of BMI SDS on peak GH values in children. It showed a significant negative relationship. Importantly, this relationship was already present in the normal range of BMI SDS and could lead to overdiagnosis of GHD in children with overweight and obesity. With the ever-rising prevalence of pediatric obesity, there is a need for BMI (SDS)-specific cutoff values for GH stimulation tests in children. Based on the evidence from this meta-analysis, we suggest the following weight status-adjusted cutoffs for GH stimulation tests that have cutoffs for children with normal weight of 5, 7, 10, and 20 µg/L: for overweight children: 4.6, 6.5, 9.3, and 18.6 µg/L; and for children with obesity: 4.3, 6.0, 8.6, and 17.3 µg/L.


The NCGENES project: exploring the new world of genome sequencing.
Massively parallel sequencing (MPS) is now a clinical reality, promising improved diagnosis, targeted therapies, and population-based screening. To realize the potential of genomics, we must learn how to apply this technology optimally. The NCGENES project is designed to address several challenges that must be overcome in order to integrate MPS into clinical care.


Label Adherence of Direct Oral Anticoagulants Dosing and Clinical Outcomes in Patients With Atrial Fibrillation.
Background Dose adjustment of non-vitamin K antagonist oral anticoagulants (NOACs) is indicated in some patients with atrial fibrillation (AF), based on selected patient factors or concomitant medications. We assessed the frequency of label adherence of NOAC dosing among AF patients and the associations between off-label NOAC dosing and clinical outcomes. Methods and Results We evaluated 53 649 AF patients treated with an NOAC using Korean National Health Insurance Service database during the period from 2013 to 2016. NOAC doses were classified as either underdosed or overdosed, consistent with Korea Food and Drug Administration labeling. Cox proportional hazards regression was performed to investigate the effectiveness and safety outcomes including stroke or systemic embolism, major bleeding, and all-cause mortality. Overall, 16 757 NOAC-treated patients (31.2%) were underdosed, 4492 were overdosed (8.4%), and 32 400 (60.4%) were dosed appropriately according to drug labeling. Compared with patients with label adherence, those who were underdosed or overdosed were older (aged 71±8 and 75±7 years versus 70±9 years, respectively; P<0.001) and had higher CHA2DS2-VASc scores (4.6±1.7 and 5.3±1.7 versus 4.5±1.8, respectively; P<0.001). NOAC overdosing was associated with increased risk for stroke or systemic embolism (5.76 versus 4.03 events/100 patient-years, P<0.001), major bleeding (4.77 versus 2.94 events/100 patient-years, P<0.001), and all-cause mortality (5.43 versus 3.05 events/100 patient-years, P<0.001) compared with label-adherent use. Conclusions In real-world practice, a significant proportion (almost 2 in 5) of AF patients received NOAC doses inconsistent with drug labeling. NOAC overdosing is associated with worse clinical outcomes in Asian AF patients.


Amphetamine actions at the serotonin transporter rely on the availability of phosphatidylinositol-4,5-bisphosphate.
Nerve functions require phosphatidylinositol-4,5-bisphosphate (PIP2) that binds to ion channels, thereby controlling their gating. Channel properties are also attributed to serotonin transporters (SERTs); however, SERT regulation by PIP2 has not been reported. SERTs control neurotransmission by removing serotonin from the extracellular space. An increase in extracellular serotonin results from transporter-mediated efflux triggered by amphetamine-like psychostimulants. Herein, we altered the abundance of PIP2 by activating phospholipase-C (PLC), using a scavenging peptide, and inhibiting PIP2-synthesis. We tested the effects of the verified scarcity of PIP2 on amphetamine-triggered SERT functions in human cells. We observed an interaction between SERT and PIP2 in pull-down assays. On decreased PIP2 availability, amphetamine-evoked currents were markedly reduced compared with controls, as was amphetamine-induced efflux. Signaling downstream of PLC was excluded as a cause for these effects. A reduction of substrate efflux due to PLC activation was also found with recombinant noradrenaline transporters and in rat hippocampal slices. Transmitter uptake was not affected by PIP2 reduction. Moreover, SERT was revealed to have a positively charged binding site for PIP2. Mutation of the latter resulted in a loss of amphetamine-induced SERT-mediated efflux and currents, as well as a lack of PIP2-dependent effects. Substrate uptake and surface expression were comparable between mutant and WT SERTs. These findings demonstrate that PIP2 binding to monoamine transporters is a prerequisite for amphetamine actions without being a requirement for neurotransmitter uptake. These results open the way to target amphetamine-induced SERT-dependent actions independently of normal SERT function and thus to treat psychostimulant addiction.


Effect of proliferative state on glutathione S-transferase isoenzyme expression in cultured rat liver epithelial cells.
A specific biochemical phenotype is expressed during chemical hepatocarcinogenesis, which includes increased activity of the various isoenzyme forms of glutathione S-transferase (GST) composed of class alpha (Ya/Yc), class mu (Yb) and class pi (Yp) gene products. In vitro cell lines of normal and chemically transformed rat liver epithelial cells provide an opportunity to examine the regulation of expression of GST isoenzymes. We have studied the effect of the state of proliferation in culture on both the enzymic activity and the isoenzyme-specific mRNA expression. In normal rat liver epithelial cells (WB-F344), basal expression of the Yp subunit decreases, and of the Yb subunit increases, in cells at confluence compared with those in logarithmic-phase growth. In a subline of WB-F344 cells that has been chemically treated in vitro (GN6), there was greater Yp expression; however, the effect of growth status on both subunits was the same as in the nontransformed WB-344 cells, and the Ya subunit was not expressed. Inhibition of RNA synthesis with actinomycin D was limited, demonstrating pronged half-lives of the GST mRNAs, and shows a slightly greater decrease in GST-Yp specific mRNA levels in the confluent cells. Also nuclear run-off experiments demonstrate identical transcription rate in confluent and pre-confluent cells. These data suggest that the increase in Yp steady-state RNA in preconfluent cells is due to increased stability of the GST-Yp mRNA. In tumor cells derived from GN6 cells, the regulatory effects of growth status on the Yb and Yp expressions were absent. However, in these cells Ya subunit was expressed and this was subject to the effects of cell proliferation. We conclude that the growth status of cells in culture exerts a significant regulatory control on the GST isoenzyme gene expression. The effects are probably mediated at independent regulatory sites in each gene. The state of transformation of rat liver epithelial cells may determine responsiveness to this effect.


The effect of nonsteroidal antiinflammatory drugs ibuprofen, flurbiprofen, and diclofenac on in vitro and in vivo growth of mouse fibrosarcoma.
For suppression of primary G:5:113 fibrosarcoma growth, three structurally different cyclooxygenase (COX) inhibitors (ibuprofen, flurbiprofen, and diclofenac) were administered intraperitoneally (i.p.) in two regimens starting on day 5 after tumor-cell inoculation. Repeated application of 0.15 mg/mouse/day during 14 consecutive days significantly suppressed the tumor growth and increased the percentage of surviving mice. Similar tendency, however without significant differences, was observed when animals were given 0.5 mg/day for five consecutive days. These results suggest that a time schedule of drug application is important for the therapeutic effect. Suppressive effect of diclofenac and flurbiprofen on tumor growth was also observed under in vitro conditions. We conclude that suppressive effect of these drugs on tumor growth in vivo comprises both direct effects of COX inhibitors on fibrosarcoma cells and indirect effects that are presumably mediated by extratumoral sources. Our findings encourage the use of COX inhibitors in the therapy of fibrosarcoma.


In situ identification of activated Ta1+ T lymphocytes in human leprosy skin lesions.
Using a monoclonal antibody, anti-Ta1, that identifies antigen-activated T lymphocytes in vitro, we sought to identify activated T lymphocytes in leprosy skin lesions. Greater numbers of Ta1 positive T lymphocytes were observed in tuberculoid leprosy, lepromin skin tests, and reversal reactions as compared with lepromatous leprosy or erythema nodosum leprosum (ENL) (p less than 0.001). With a double-staining technique, we found that the majority of these activated T lymphocytes were of the helper/inducer phenotype. No differences of Ta1 positive lymphocytes were observed in the peripheral blood. The defective cell-mediated immune response in lepromatous and ENL patients correlates with, and may be related to the failure of T-helper/inducer activation or proliferation in the presence of Mycobacterium leprae.


Fluorescence lifetime spectroscopic imaging with measurements of photon migration.
Frequency-domain measurements of photon migration are coupled with a model of fluorescence generation and propagation in order to develop a method for reconstructing maps of fluorescent properties within interior tissue volumes from exterior measurements at the air-tissue interface. Simulation results confirm the feasibility of optical imaging through the use of exogenously administered contrast agents on the basis of fluorophore decay kinetics and yield. Experimental measurements using single-pixel and multipixel devices illustrate that the contrast owing to exogenous fluorescence exceeds that owing to absorption or scattering caused by endogenous chromophores or tissue structure and owing to absorption caused by exogenous contrast agents.


Rapid and Selective Biomarker Detection with Conductometric Sensors.
The biomarker detection in human body fluids is crucial as biomarkers are important in diagnosing diseases. Conventional invasive techniques for biomarker detection are associated with infection, tissue damage, and discomfort. Non-invasive devices are an attractive alternative. Here, metal oxide (oxygen-deficient zinc oxide, ZnO) based conductometric sensors with two-terminal electrodes for rapid detection of biomarkers in real-time, are presented. This platform can be engineered for non-invasive, sensitive, and on-demand selective detection of biomarkers based on surface functionalization. The three novelties in this biosensing technique include an on-demand target selection device platform, short (<10 min) incubation times, and real-time monitoring of the biomarker of interest by electrical (resistance change) measurements. Cardiac inflammatory biomarkers interleukin 6 (IL-6) and C-reactive protein (CRP) are used as the model antigens. The devices can detect 100× lower concentration of IL-6 than healthy levels in human saliva and sweat and 1000× and ≈50× lower CRP concentrations than healthy levels in human saliva and sweat, respectively. The devices show high selectivity for IL-6 and CRP antigens when tested with a mixture of biomarkers. This sensor platform can be extended to selective measurements for viruses or DNA screening, which enables a new category of compact and rapid point-of-care medical devices.


To Eat and to Be Eaten: Mutual Metabolic Adaptations of Immune Cells and Intracellular Bacterial Pathogens upon Infection.
Intracellular bacterial pathogens (IBPs) invade and replicate in different cell types including immune cells, in particular of the innate immune system (IIS) during infection in the acute phase. However, immune cells primarily function as essential players in the highly effective and integrated host defense systems comprising the IIS and the adaptive immune system (AIS), which cooperatively protect the host against invading microbes including IBPs. As countermeasures, the bacterial pathogens (and in particular the IBPs) have developed strategies to evade or reprogram the IIS at various steps. The intracellular replication capacity and the anti-immune defense responses of the IBP's as well as the specific antimicrobial responses of the immune cells of the innate and the AIS depend on specific metabolic programs of the IBPs and their host cells. The metabolic programs of the immune cells supporting or counteracting replication of the IBPs appear to be mutually exclusive. Indeed, recent studies show that upon interaction of naïve, metabolically quiescent immune cells with IBPs, different metabolic activation processes occur which may result in the provision of a survival and replication niche for the pathogen or its eradication. It is therefore likely that within a possible host cell population subsets exist that are metabolically programmed for pro- or anti-microbial conditions. These metabolic programs may be triggered by the interactions between different bacterial agonistic components and host cell receptors. In this review, we summarize the current status in the field and discuss metabolic adaptation processes within immune cells of the IIS and the IBPs that support or restrict the intracellular replication of the pathogens.


Does child verbal ability mediate the relationship between maternal sensitivity and later self-regulation? A longitudinal study from infancy to 4 years.
There is a need to further examine the mechanisms by which maternal sensitivity influences the development of child self-regulation. This study investigated the role of maternal sensitivity when infants were 10 months old and child verbal ability at 18 months, in relation to various aspects of self-regulation at 48 months, in a sample of 95 typically developing children (46.3% girls). In particular, the study examined, from a Vygotskian perspective, whether child verbal ability, as measured by receptive and expressive language, mediated the relationship between maternal sensitivity and hot and cool aspects of self-regulation in the child. As hypothesized, maternal sensitivity predicted child verbal ability, as well as working memory, set shifting, and delay of gratification. Child receptive language predicted set shifting, inhibition, and delay of gratification. In addition, receptive language mediated the relationship between maternal sensitivity and inhibition only. Additive effects of maternal sensitivity and child receptive language in relation to set shifting were found, and a main effect of maternal sensitivity on child delay of gratification. The results add to the body of research suggesting that responsive parenting and child verbal ability are important for the development of self-regulation, and suggest that different mechanisms may be at work for different aspects of self-regulation.


Noninvasive assessment of human jawbone using ultrasonic guided waves.
The problem of detecting defects in jawbones is an important problem. Existing methods based on Xrays are invasive and constrain the achievable image quality. They also may carry known risks of cancer generation or may be limited in accurate diagnosis scope. This work is motivated by the lack of current imaging modalities to accurately predict the mechanical properties and defects in jawbone. Ultrasonic guided waves are sensitive to changes in microstructural properties and thus have been widely used for noninvasive material characterization. Using these waves may provide means for early diagnosis of marrow ischemic disorders via detecting focal osteoporotic marrow defect, chronic nonsuppurative osteomyelitis, and cavitations in the mandible (jawbone). Guided waves propagating along the mandibles may exhibit dispersion behavior that depends on material properties, geometry, and embedded cavities. In this work, we present the first study in the theoretical and experimental analysis of guided wave propagation in jawbone. Semianalytical, finite-element (SAFE) method is used to analyze dispersion behavior of guided waves propagating in human mandibles. The geometry of the cross section is obtained by segmenting the computed tomography (CT) images of the jawbone. The cross section of the mandible is divided in two regions representing the cortical and trabecular bones. Each region is modeled as a linear Hookean material. The material properties for both regions are adopted from the literature. The experimental setup for the guided waves experiment is described. The results from both numerical analysis and guided waves experiment exhibit variations in the group velocity of the first arrival signal and in the dispersion behavior of healthy and defected mandibles. These results shall provide a means to noninvasively characterize the jawbone and accurately assess the bone mechanical properties. Our study is not aimed at characterizing the bone density in human mandibles. Rather, it is aimed to assess bone mechanical properties and defects that cannot be diagnosed by X-ray or other imaging modalities. This work may pave the way to the development of inexpensive noninvasive devices to detect small defects in human mandibles.


The effects of wastewater effluent on multiple behaviours in the amphipod, Gammarus pulex.
The prevalence of pharmaceuticals and personal care products (PPCPs) in lotic habitats is increasing, with the main source of these contaminants being effluent from waste water treatment works (WwTW). There is still much uncertainty about the impacts of these PPCPs at environmentally relevant concentrations and their potential effects on aquatic ecology. Behaviour is a sensitive endpoint which can help evaluate possible population level effects from changes in physiology. This paper evaluates the effects of WwTW effluent on a range of behaviours in the freshwater invertebrate, Gammarus pulex. Effluent taken from the outflow of two WwTW in southern England was used in the study. Behavioural analyses, namely feeding rate, phototaxis, activity, velocity and precopula pairing, were measured in G. pulex following a period of one and three weeks after exposure to a 50% or 100% effluent and a control. Mortality remained very low throughout the 3 week experiment (0-10%, n = 20) and no significant changes in moulting frequency were observed (p > 0.05). No significant effects on feeding or velocity or phototaxis following 3 weeks of effluent exposures were observed (p > 0.05). However, significant reductions were observed in the overall activity over 3 weeks across which appeared to be exacerbated by exposure to effluents. Interestingly, males exposed for 3 weeks to WwTW effluent re-paired with unexposed females significantly faster (4-6x) than control animals. This result was consistent between the effluents taken from the two WwTW. The implications of these behavioural changes are currently unknown but highlight the need for a varied set of tools to study the behavioural changes in wildlife.


Identifying pathogenicity of human variants via paralog-based yeast complementation.
To better understand the health implications of personal genomes, we now face a largely unmet challenge to identify functional variants within disease-associated genes. Functional variants can be identified by trans-species complementation, e.g., by failure to rescue a yeast strain bearing a mutation in an orthologous human gene. Although orthologous complementation assays are powerful predictors of pathogenic variation, they are available for only a few percent of human disease genes. Here we systematically examine the question of whether complementation assays based on paralogy relationships can expand the number of human disease genes with functional variant detection assays. We tested over 1,000 paralogous human-yeast gene pairs for complementation, yielding 34 complementation relationships, of which 33 (97%) were novel. We found that paralog-based assays identified disease variants with success on par with that of orthology-based assays. Combining all homology-based assay results, we found that complementation can often identify pathogenic variants outside the homologous sequence region, presumably because of global effects on protein folding or stability. Within our search space, paralogy-based complementation more than doubled the number of human disease genes with a yeast-based complementation assay for disease variation.


Resuscitation with Pooled and Pathogen-Reduced Plasma Attenuates the Increase in Brain Water Content following Traumatic Brain Injury and Hemorrhagic Shock in Rats.
Traumatic brain injury and hemorrhagic shock is associated with blood-brain barrier (BBB) breakdown and edema formation. Recent animal studies have shown that fresh frozen plasma (FFP) resuscitation reduces brain swelling and improves endothelial function compared to isotonic NaCl (NS). The aim of this study was to investigate whether pooled and pathogen-reduced plasma (OctaplasLG® [OCTA]; Octapharma, Stockholm, Sweden) was comparable to FFP with regard to effects on brain water content, BBB permeability, and plasma biomarkers of endothelial glycocalyx shedding and cell damage. After fluid percussion brain injury, hemorrhage (20 mL/kg), and 90-min shock, 48 male Sprague-Dawley rats were randomized to resuscitation with OCTA, FFP, or NS (n = 16/group). Brain water content (wet/dry weight) and BBB permeability (transfer constant for 51Cr-EDTA) were measured at 24 h. Plasma osmolality, oncotic pressure, and biomarkers of systemic glycocalyx shedding (syndecan-1) and cell damage (histone-complexed DNA) were measured at 0 and 23 h. At 24 h, brain water content was 80.44 ± 0.39%, 80.82 ± 0.82%, and 81.15 ± 0.86% in the OCTA, FFP, and NS groups (lower in OCTA vs. NS; p = 0.026), with no difference in BBB permeability. Plasma osmolality and oncotic pressures were highest in FFP and OCTA resuscitated, and osmolality was further highest in OCTA versus FFP (p = 0.027). In addition, syndecan-1 was highest in FFP and OCTA resuscitated (p = 0.010). These results suggest that pooled solvent-detergent (SD)-treated plasma attenuates the post-traumatic increase in brain water content, and that this effect may, in part, be explained by a high crystalloid and colloid osmotic pressure in SD-treated plasma.


Fucoxanthin inhibits tumour-related lymphangiogenesis and growth of breast cancer.
Tumour lymphangiogenesis plays an important role in promoting the growth and lymphatic metastasis of tumours. The process is associated with cell proliferation, migration and tube-like structure formation in lymphatic endothelial cells (LEC), but no antilymphangiogenic agent is currently used in clinical practice. Fucoxanthin is a material found in brown algae that holds promise in the context of drug development. Fucoxanthin is a carotenoid with variety of pharmacological functions, including antitumour and anti-inflammatory effects. The ability of fucoxanthin to inhibit lymphangiogenesis remains unclear. The results of experiments performed as part of this study show that fucoxanthin, extracted from Undaria pinnatifida (Wakame), inhibits proliferation, migration and formation of tube-like structures in human LEC (HLEC). In this study, fucoxanthin also suppressed the malignant phenotype in human breast cancer MDA-MB-231 cells and decreased tumour-induced lymphangiogenesis when used in combination with a conditional medium culture system. Fucoxanthin significantly decreased levels of vascular endothelial growth factor (VEGF)-C, VEGF receptor-3, nuclear factor kappa B, phospho-Akt and phospho-PI3K in HLEC. Fucoxanthin also decreased micro-lymphatic vascular density (micro-LVD) in a MDA-MB-231 nude mouse model of breast cancer. These findings suggest that fucoxanthin inhibits tumour-induced lymphangiogenesis in vitro and in vivo, highlighting its potential use as an antilymphangiogenic agent for antitumour metastatic comprehensive therapy in patients with breast cancer.


Embryology of myelomeningocele and anencephaly.
The authors review current views on of the embryogenesis of the neural tube defects (NTDs) myelomeningocele and anencephaly. In this context, the following four approaches to the study of NTDs are discussed: normal morphogenesis and timing of early human neural development from conception to the ascent of the conus medullaris; mechanical and molecular biology of neural tube closure derived from experimental and animal models; morphological and biomechanical features of the NTDs myelomeningocele and anencephaly; and the experimental evidence for the importance of both genetic and environmental influences on human NTDs. Although considerable insight into both normal neural tube closure and the factor(s) by which this process may be disrupted has been reported in recent years, the exact mechanism(s) by which human myelomeningoceles and anencephaly arise remain elusive.


Exact solutions for evolutionary strategies on harmonic landscapes.
In this paper two different evolutionary strategies are tested by means of harmonic landscapes. Both strategies are based on ensembles of searchers, spreading over the search space according to laws inspired by nature. The main difference between the two prototypes is given by the underlying selection mechanism, governing the increase or decrease of the local population of searchers in certain regions of the search space. More precisely, we compare a thermodynamic strategy, which is based on a physically motivated local selection criterion, with a biologically motivated strategy, which features a global selection scheme (i.e., global coupling of the searchers). Confining ourselves to a special class of initial conditions, we show that, in the simple case of harmonic test potentials, both strategies possess particular analytical solutions of the same type. By means of these special solutions, the velocities of the two strategies can be compared exactly. In the last part of the paper, we extend the scope of our discussion to a mixed strategy, combining local and global selection.


Impact of Centralisation on Abdominal Aortic Aneurysm Repair Outcomes: Early Experience in Catalonia.
Several studies have revealed high volume centres have better outcomes in the treatment of abdominal aortic aneurysms (AAAs), thus supporting centralisation of this procedure into selected centres based on volume. To date however, the real benefit of centralisation of this pathology has not been well demonstrated. The aim of this study was to analyse the impact of centralisation in to high volume centres (defined as those performing more than 30 cases per year) on AAA treatment outcomes carried out in Catalonia (Spain). Data were collected from official national registries (HDMBD) for AAA treated by endovascular aneurysm repair (EVAR) or open repair (OR) over a nine year period. Two time periods were selected for comparison: before centralisation (2009-2014) and after complete centralisation (2015-2017). The primary objective was to determine short term mortality (in hospital and 30 day mortality) and length of stay (LOS) after intact AAA (iAAA) and ruptured AAA (rAAA) repair, before and after centralisation. Uni- and multivariable analyses were performed in order to identify independent outcomes predictors. A total of 3 501 iAAAs, including 1 124 (32.1%) OR and 2377 (67.9%) EVAR, and 409 rAAAs, including 218 (53.3%) OR and 191 (46.7%) EVAR, were identified. After centralisation, there was a significant decrease in overall mortality in iAAA repair (4.7% vs. 2.0%, p < .001) and rAAA repair (53.1% vs. 41.9%, p = .028). Mortality reduction in iAAAs was significant for OR (8.7% vs. 3.6%, p = .005), but not for EVAR (2.2% vs. 1.5%, p = .25). Overall LOS decreased as well, mainly in iAAAs (9.49 ± 10.84 vs. 7.44 ± 12.23 days, p < .001), and in particular in elective EVAR (7.32 ± 7.73 vs. 6.00 ± 8.97 days, p < .001). Multivariable analysis was identified before the centralisation period as an independent predictor for both mortality (odds ratio 1.484, 95% CI 1.098-2.005, p = .010) and LOS (B coefficient 1.146, 95% CI 0.218-2.073, p = .016). The implementation of a country based centralisation programme for AAA treatment led to a significant reduction in short term mortality, for both iAAA and rAAA, and mainly for elective OR. LOS also significantly decreased, mainly for elective EVAR. These results support the benefit of centralisation of AAA repair procedures.


Nrf2-dependent antioxidant response mediated the protective effect of tanshinone IIA on doxorubicin-induced cardiotoxicity.
Doxorubicin (DOX), a potent and widely used anticancer agent, can give rise to severe cardiotoxicity that limits its clinical use by inducing oxidative stress. Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is the central regulator of cellular responses to electrophilic/oxidative stress, which serves a critical role in maintenance of normal cardiac function. Tanshinone IIA (Tan IIA) has previously been reported to protect against DOX-induced cardiotoxicity. The aim of the present study was to elucidate whether Nrf2 signaling serves a role in the underlying mechanism. In the animal model, DOX induced acute cardiotoxicity, whereas Tan IIA pretreatment reduced the activity of myocardial enzymes, and increased activity of the antioxidant enzymes superoxide dismutase, catalase and glutathione (GSH). Furthermore, Tan IIA pretreatment (3-10 µM) significantly increased the cell viability and markedly restored morphological changes in DOX-injured H9c2 cells, decreased the generation of reactive oxygen species, and increased the level of intracellular GSH. Additionally, Tan IIA pretreatment also induced the nuclear accumulation of Nrf2 and its downstream genes heme oxygenase-1, NAD(P)H dehydrogenase (quinone) 1, and glutamate-cysteine ligase catalytic subunit in both the mice cardiac tissues and H9c2 cells. Nrf2 knockdown by small interfering RNA downregulated Tan IIA-induced Nrf2 activation and reversed the effect of Tan IIA on the DOX-induced inhibition of cell viability. These results suggest that the Nrf2-dependent antioxidant response mediates the protective effect of Tan IIA on DOX-induced cardiotoxicity.


Structure of a psoralen cross-linked DNA in solution by nuclear magnetic resonance.
One- and two-dimensional nuclear magnetic resonance (NMR) methods were used to determine a three-dimensional model of an eight-base-pair DNA fragment (d-GGGTACCC) cross-linked with psoralen in solution. Two-dimensional nuclear Overhauser effect experiments were used to assign the spectrum and estimate distances for 171 proton pairs in the cross-linked DNA. The NMR-derived model shows a 53 degree bend into the major groove that occurs primarily at the site of drug addition and a 56 degree unwinding that spans the eight-base-pair duplex.


Contactin 2 homophilic adhesion structure and conformational plasticity.
The cell-surface attached glycoprotein contactin 2 is ubiquitously expressed in the nervous system and mediates homotypic cell-cell interactions to organize cell guidance, differentiation, and adhesion. Contactin 2 consists of six Ig and four fibronectin type III domains (FnIII) of which the first four Ig domains form a horseshoe structure important for homodimerization and oligomerization. Here we report the crystal structure of the six-domain contactin 2Ig1-6 and show that the Ig5-Ig6 combination is oriented away from the horseshoe with flexion in interdomain connections. Two distinct dimer states, through Ig1-Ig2 and Ig3-Ig6 interactions, together allow formation of larger oligomers. Combined size exclusion chromatography with multiangle light scattering (SEC-MALS), small-angle X-ray scattering (SAXS) and native MS analysis indicates contactin 2Ig1-6 oligomerizes in a glycan dependent manner. SAXS and negative-stain electron microscopy reveals inherent plasticity of the contactin 2 full-ectodomain. The combination of intermolecular binding sites and ectodomain plasticity explains how contactin 2 can function as a homotypic adhesion molecule in diverse intercellular environments.


Electromyographic Analysis of Paraspinal Muscles of Scoliosis Patients Using Machine Learning Approaches.
A large number of studies have used electromyography (EMG) to measure the paraspinal muscle activity of adolescents with idiopathic scoliosis. However, investigations on the features of these muscles are very limited even though the information is useful for evaluating the effectiveness of various types of interventions, such as scoliosis-specific exercises. The aim of this cross-sectional study is to investigate the characteristics of participants with imbalanced muscle activity and the relationships among 13 features (physical features and EMG signal value). A total of 106 participants (69% with scoliosis; 78% female; 9-30 years old) are involved in this study. Their basic profile information is obtained, and the surface EMG signals of the upper trapezius, latissimus dorsi, and erector spinae (thoracic and erector spinae) lumbar muscles are tested in the static (sitting) and dynamic (prone extension position) conditions. Then, two machine learning approaches and an importance analysis are used. About 30% of the participants in this study find that balancing their paraspinal muscle activity during sitting is challenging. The most interesting finding is that the dynamic asymmetry of the erector spinae (lumbar) group of muscles is an important (third in importance) predictor of scoliosis aside from the angle of trunk rotation and height of the subject.


Cephalometric predictors for optimal soft tissue profile outcome in adult Asian class I subjects treated via extraction and non-extraction. A retrospective study.
The aim of this retrospective study was to identify cephalometric predictors associated with favourable soft tissue profile outcomes after premolars extraction and non-extraction in class I malocclusion subjects. A total of 80 subjects, treated with non-extraction and premolars extraction (40 subjects each), were equally divided into favourable (FG) and unfavourable (UFG) groups using subjective and objective soft tissue profile outcome assessment methods. An independent t-test was utilized for the comparison of cephalometric measurements between the non-extraction (NE) and premolars extraction (PME) treatment modalities. Cox proportional hazard algorithm regression analysis was performed to identify cephalometric factors associated with favourable soft tissue outcomes. The pre-treatment mean age of the NE group was 20.2±2.3 and PME group was 20.2±2.5 years. After dividing the sample of the NE and PME groups according to subjective and objective soft-tissue outcome assessment criteria, FG and UFG consisted of 20 subjects each. Cox proportional hazard algorithm regression analysis found upper incisor to NA angle (95% CI: 1.033, 1.196) to be associated with FG in NE and upper incisor to SN (95% CI: 1.018, 1.206) and ANB angle (95% CI:1.165, 3.608) in PME. There was a statistically significant strong correlation between subjective and objective evaluation methods (P≤0.001). Cephalometric analysis is a valuable tool to predict soft-tissue outcomes after NE and PME. Increased upper incisors inclinations at the start of NE treatment result in favourable soft tissue profile outcomes. Slightly convex profile and proclined maxillary incisors are the predictors of favourable soft tissue profile outcome after PME. There was a statistically significant association between subjective and objective evaluation criteria of soft tissue outcomes.


Exploration of the role of oxidative stress-related genes in LPS-induced acute lung injury via bioinformatics and experimental studies.
During the progression of acute lung injury (ALI), oxidative stress and inflammatory responses always promote each other. The datasets analyzed in this research were acquired from the Gene Expression Omnibus (GEO) database. The Weighted Gene Co-expression Network Analysis (WGCNA) and limma package were used to obtain the ALI-related genes (ALIRGs) and differentially expressed genes (DEGs), respectively. In total, two biological markers (Gch1 and Tnfaip3) related to oxidative stress were identified by machine learning algorithms, Receiver Operator Characteristic (ROC), and differential expression analyses. The area under the curve (AUC) value of biological markers was greater than 0.9, indicating an excellent power to distinguish between ALI and control groups. Moreover, 15 differential immune cells were selected between the ALI and control samples, and they were correlated to biological markers. The transcription factor (TF)-microRNA (miRNA)-Target network was constructed to explore the potential regulatory mechanisms. Finally, based on the quantitative reverse transcription polymerase chain reaction (qRT-PCR), the expression of Gch1 and Tnfaip3 was significantly higher in ALI lung tissue than in healthy controls. In conclusion, the differences in expression profiles between ALI and normal controls were found, and two biological markers were identified, providing a research basis for further understanding the pathogenesis of ALI.


A rare case of bleeding in a cerebellopontine angle epidermoid cyst.
Epidermoid cysts of the central nervous system are described as rare, benign, slow growing lesions with a history of high rate of recurrence even after surgical removal. This lesion is rarely located at the cerebellopontine angle and is found to be composed of solid and cystic components with close adherence to vital neurovascular structures that might complicate its removal. We present a rare case of a twenty-five year old housewife with signs and symptoms of increased intracranial pressure due to the above pathology after multiple episodes of intra-tumoral bleeding. Microneurosurgical techniques were used for tumour dissection and excision. The patient recovered well after a three-year follow-up. This is the fourth example in the index medicus of bleeding seen in a histopathologically proven cerebellopontine angle intraepidermoid cyst.


B/N-Enriched Semi-Conductive Polymer Film for Micro-Supercapacitors with AC Line-Filtering Performance.
Microsupercapacitors (MSCs) have drawn great attention for use as miniaturized electrochemical energy storage devices in portable, wearable, as well as implantable electronics. Many materials have been developed as electrodes for MSCs. However, the thin-film fabrication for most of these materials involves multistep operations, including filtration, spray coating, and sputtering. Most importantly, these methods present challenges for the preparation of thin films at the atomic or molecular scale. Therefore, the understanding of performance of ultrathin-film-based MSCs remains challenge. Herein, a B/N-enriched polymer film is successfully prepared using the photoassisted interfacial approach. The as-synthesized polymer film exhibits typical semiconductive characteristics and can be easily scaled up to a large area of up to tens of square centimeters. This ultrathin polymer film can be directly transferred to silicon wafers to fabricate MSC through laser scribing. The prepared MSC exhibits specific volumetric capacitance as high as 20.9 F cm-3, corresponding to volumetric energy density of 2.9 mWh cm-3 (at 0.1 V s-1). Moreover, the volumetric power density can reach 1461 W cm-3, surpassing most existing semiconductive polymer film-based MSC devices. In addition, the prepared MSC exhibits typical AC line-filtering ability (-67° at 120 Hz). This study offers a facile interfacial approach to preparing semiconductive polymer films with aromatic moieties for microsized energy storage devices.


The Vanadium Iodoperoxidase from the marine flavobacteriaceae species Zobellia galactanivorans reveals novel molecular and evolutionary features of halide specificity in the vanadium haloperoxidase enzyme family.
Vanadium haloperoxidases (VHPO) are key enzymes that oxidize halides and are involved in the biosynthesis of organo-halogens. Until now, only chloroperoxidases (VCPO) and bromoperoxidases (VBPO) have been characterized structurally, mainly from eukaryotic species. Three putative VHPO genes were predicted in the genome of the flavobacterium Zobellia galactanivorans, a marine bacterium associated with macroalgae. In a phylogenetic analysis, these putative bacterial VHPO were closely related to other VHPO from diverse bacterial phyla but clustered independently from eukaryotic algal VBPO and fungal VCPO. Two of these bacterial VHPO, heterogeneously produced in Escherichia coli, were found to be strictly specific for iodide oxidation. The crystal structure of one of these vanadium-dependent iodoperoxidases, Zg-VIPO1, was solved by multiwavelength anomalous diffraction at 1.8 Å, revealing a monomeric structure mainly folded into α-helices. This three-dimensional structure is relatively similar to those of VCPO of the fungus Curvularia inaequalis and of Streptomyces sp. and is superimposable onto the dimeric structure of algal VBPO. Surprisingly, the vanadate binding site of Zg-VIPO1 is strictly conserved with the fungal VCPO active site. Using site-directed mutagenesis, we showed that specific amino acids and the associated hydrogen bonding network around the vanadate center are essential for the catalytic properties and also the iodide specificity of Zg-VIPO1. Altogether, phylogeny and structure-function data support the finding that iodoperoxidase activities evolved independently in bacterial and algal lineages, and this sheds light on the evolution of the VHPO enzyme family.


Bilophila wadsworthia: a unique Gram-negative anaerobic rod.
Although comprising less than 0.01% of the normal human gastrointestinal microbiota, Bilophila wadsworthia is the third most common anaerobe recovered from clinical material obtained from patients with perforated and gangrenous appendicitis. Since its discovery in 1988, B. wadsworthia has been recovered from clinical specimens associated with a variety of infections, including sepsis, liver abscesses, cholecystitis, Fournier's gangrene, soft tissue abscesses, empyema, osteomyelitis, Bartholinitis, and hidradenitis suppurativa. In addition, it has been found in the saliva and vaginal fluids of asymptomatic adults and even in the periodontal pockets of dogs. The organism is a saccharolytic, fastidious, and is easily recognized by its strong catalase reaction with 15% H2O2, production of hydrogen sulfide, and growth stimulation by bile (oxgall) and pyruvate. Approximately 75% of strains are urease positive. When grown on pyruvate-containing media, > 85% of strains demonstrate beta-lactamase production. Ribosomal RNA-based phylogenetic studies show Bilophila to be a homogeneous species, most closely related to Desulfovibrio species. Both adherence to human cells and endotoxin have been observed, and preliminary work suggests that environmental iron has a role in expression of outer membrane proteins. Penicillin-binding proteins appear to mediate the organism's susceptibility to at least some beta-lactam agents, which induce spheroplast formation that results in a haze of growth on agar dilution susceptibility test plates which is difficult to interpret. Bilophilastrains are inhibited in vitro by most antibiotics.


A pilot study to assess a risk of a high-risk group of low back pain membership in workers who perform the manual material handling tasks.
We conducted this experimental study to estimate a risk of a high-risk group of low back pain (LBP) membership in workers who perform the manual material handling (MMH) tasks in an actual workplace setting. The subjects include healthy workers who were engaged in 12 MMH tasks at 6 manufacturing companies. We assessed the dynamic motion of trunk or lumbar spine using an industrial lumbar motion monitor (BioDynamics Laboratory of Ohio State University). The subjects were evaluated for the age, gender, years of working and anthropometric measurements (e.g., height, weight, shoulder height, elbow height, iliac height, leg length, trunk length, trunk circumference, iliac width, iliac depth, xiphoid width and xiphoid depth). Moreover, they were also evaluated for a risk of a high-risk group of LBP membership based on lift frequency, average twisting velocity, maximum moment, maximum sagittal flexion and maximum lateral velocity. The subjects who were engaged in a packaging at a detergent manufacturing company are at the greatest risk of LBP (63.76%). This was followed by packaging at a leather product manufacturing company (57.06%), packaging at a non-metallic casting material manufacturing company (57.03%), manual injection at a non-metallic casting material manufacturing company (52.00%), toggling at a leather product manufacturing company (46.09%), non-metallic casting material manufacturing company (42.88%), rolling at a non-metallic mineral product manufacturing company (42.12%), shooting at a non-metallic casting material manufacturing company (40.99%), vacuum processes at a leather product manufacturing company (35.00%), looping at a general industrial machinery manufacturing company (33.93%), setting at a leather product manufacturing company (30.22%) and packaging at a general metal product manufacturing company (22.02%). Our approach indicates that there is a risk of a high-risk group of LBP membership in workers who perform the MMH tasks.


Change of middle ear transfer function in otitis media with effusion model of guinea pigs.
Otitis media with effusion (OME) is an inflammatory disease of the middle ear that causes most cases of conductive hearing loss observed in the pediatric population. With the long term goal of evaluating middle ear function with OME, the aim of the current study was to create an animal model of OME in which middle ear transfer functions could be measured. In guinea pigs, OME was created by injecting lipopolysaccharide (LPS) into the middle ear. Evidence of OME was assessed by otoscopy, tympanometry, histology, and by measuring the volume of fluid in the middle ear. Vibrations of the umbo and round window membrane were measured with a laser Doppler vibrometer at frequency range of 200-40 kHz in three groups of 3, 7, and 14 days after injection of LPS. Changes in displacement of the umbo and round window membrane in response to 80 dB SPL sound in the ear canal were measured across the frequency range. Displacement of both the umbo and round window membrane was reduced at all time points following LPS injections. Further, the change of the displacement transmission ratio (DTR) from the tympanic membrane to the round window occurred mainly in chronic (e.g. 14 days post-LPS injection) OME ears. This study provides useful data for analyzing the change of middle ear transfer function in OME ears.


The significance of organic carbon compounds for in situ metabolism and chemotaxis of phototrophic consortia.
The significance of organic carbon substrates for the chemotaxis and physiology of phototrophic consortia was investigated in a dense chemocline community of Pelochromatium roseum. For the first time, the monopolar monotrichous flagellation of the central bacterium could be visualized. In situ, intact motile P. roseum consortia were strongly attracted by sulphide and 2-oxoglutarate, which indicated a potential role of these compounds in the metabolism of P. roseum. In chemocline water samples, 2-[14C(U)]-oxoglutarate was utilized at nanomolar concentrations (half saturation constant of uptake Kt < or = 10-40 nM), and at a maximum uptake rate of Vmax approximately 6 nM h-1. The calculated turnover of 2-oxoglutarate at in situ concentrations was approximately 6 h. Microautoradiography of chemocline water samples revealed that 87.5% of the P. roseum consortia incorporated 2-oxoglutarate when both light and sulphide were present, whereas uptake was detected in less than 1.4% of the consortia if either light or sulphide were absent. Because the green sulphur bacterial epibionts in P. roseum have been shown to grow autotrophically, 2-oxoglutarate most likely is taken up and utilized by the central bacterium. Thus, our results indicate that incorporation of 2-oxoglutarate by the central bacterium is regulated by the metabolic state of the green sulphur bacterial epibionts.


[Choosing the method of reconstructive operation in patients with atherosclerotic occlusion of the femoral-popliteal-tibial segment in the stage of critical ischaemia].
Presented herein are remote results of surgical treatment of 101 patients with atherosclerotic occlusion of the femoral-popliteal-tibial segment in the stage of critical ischaemia. The choice of the reconstructive operation was made based on the developed criteria. Femoral-popliteal shunting into the isolated segment of the reversed great saphenous vein was performed in 48 (47.52%) patients, and 53 (52.48%) patients underwent distal femoraltibial bypass grafting according to the in situ technique. The five-year patency of the graft in these groups was identical, amounting to 54.17 and 58.49%, respectively. In order to predict the duration of bypass graft patency in the isolated segment of the popliteal artery we worked out criteria of competence of the collateral blood flow. We also defined more exactly the indications for applying a relieving arteriovenous fistula in distal femorotibial shunting. When deciding upon a method of operative treatment, preference is given to shunting into the isolated popliteal artery in sufficient collateral blood flow.