Iodination by stimulated human neutrophils. Studies on its stoichiometry, subcellular localization and relevance to microbial killing.
Myeloperoxidase of phagocytic leucocytes is thought to utilize H2O2 to oxidize halides, which then react with and kill ingested microbes. This hypothesis was based largely on the incorporation of radiolabelled iodide into cells that had phagocytosed bacteria. The present studies investigated the stoichiometry of these reactions and the subcellular localization and electrophoretic pattern of the cellular components that became iodinated. 1. The stoichiometry of the reactions are such that only a small proportion (less than 0.3%) of the total oxygen consumed is utilized for iodination. Iodination after stimulation with the soluble stimulus phorbol myristate acetate (PMA), which is not known to involve the azurophil granules and their contained myeloperoxidase, was comparable with that occurring after bacterial ingestion. 2. Analytical subcellular fractionation of cells that had phagocytosed bacteria localized about 25% of the radioactivity to the membranes, and most of the residual radioactivity distributed with the bacteria and dense granules. In cells stimulated with PMA, more of the radioactivity was associated with the membranes, but about half was still associated with the dense granules. 3. Autoradiographs after dodecyl sulphate/polyacrylamide-gel electrophoresis of cells stimulated with opsonized bacteria gave a similar distribution of iodinated components to that obtained with cells that had been stimulated with PMA or iodinated with Iodogen. These patterns of iodination were very different from those obtained when bacteria alone were iodinated with Iodogen or myeloperoxidase and H2O2. Preparations in which bacteria had been phagocytosed did not show evidence of iodination of bacterial proteins or coating opsonins. Thus positive evidence for the iodination of bacteria has not been produced, and the role of iodination in the microbicidal process of neutrophils remains to be established.


Crystal Structures of Full-Length Lanosterol 14α-Demethylases of Prominent Fungal Pathogens Candida albicans and Candida glabrata Provide Tools for Antifungal Discovery.
Targeting lanosterol 14α-demethylase (LDM) with azole drugs provides prophylaxis and treatments for superficial and disseminated fungal infections, but cure rates are not optimal for immunocompromised patients and individuals with comorbidities. The efficacy of azole drugs has also been reduced due to the emergence of drug-resistant fungal pathogens. We have addressed the need to improve the potency, spectrum, and specificity for azoles by expressing in Saccharomyces cerevisiae functional, recombinant, hexahistidine-tagged, full-length Candida albicans LDM (CaLDM6×His) and Candida glabrata LDM (CgLDM6×His) and determining their X-ray crystal structures. The crystal structures of CaLDM6×His, CgLDM6×His, and ScLDM6×His have the same fold and bind itraconazole in nearly identical conformations. The catalytic domains of the full-length LDMs have the same fold as the CaLDM6×His catalytic domain in complex with posaconazole, with minor structural differences within the ligand binding pocket. Our structures give insight into the LDM reaction mechanism and phenotypes of single-site CaLDM mutations. This study provides a practical basis for the structure-directed discovery of novel antifungals that target LDMs of fungal pathogens.


Cloning and characterization of genes involved in production of mannose-resistant, neuraminidase-susceptible (X) fimbriae from a uropathogenic O6:K15:H31 Escherichia coli strain.
The uropathogenic Escherichia coli strain 536 (O6:K15:H31) exhibits a mannose-resistant hemagglutination phenotype (Mrh) with bovine erythrocytes and delayed Mrh with human and guinea pig erythrocytes. Neuraminidase treatment of the erythrocytes abolishes mannose resistant hemagglutination, which is typical for X fimbriae. E. coli strain 536 synthesizes two different fimbriae (Fim phenotype) protein subunits, 16.5 and 22 kilodaltons in size. In addition the strain shows mannose-sensitive hemagglutination and common type I (F1) fimbriae. The cosmid clone E. coli K-12(pANN801) and another nine independently isolated Mrh+ cosmid clones derived from a cosmid gene bank of strain 536 express the 16.5-kilodalton protein band, but not the 22-kilodalton protein, indicating an association of the Mrh+ property with the "16.5-kilodalton fimbriae." All cosmid clones were fimbriated, and they reacted with antiserum produced against Mrh+ fimbriae of the E. coli strain HB101(pANN801) and lacked mannose-sensitive hemagglutination (F1) fimbriae. From the Mrh fim cosmid DNA pANN801, several subclones coding for hemagglutination and X fimbriae were constructed. Subclones that express both hemagglutination and fimbriae and subclones that only code for the hemagglutination antigen were isolated; subclones that only produce fimbriae were not detected. By transposon Tn5 mutagenesis we demonstrated that about 6.5 kilobases of DNA is required for the Mrh+ Fim+ phenotype, and the 1.5- to 2-kilobase DNA region coding for the structural protein of the fimbriae has been mapped adjacent to the region responsible for the Mrh+ phenotype. Two different regions can thus be distinguished in the adhesion determinant, one coding for hemagglutination and the other coding for fimbria formation. Transformation of plasmid DNA from these subclones into a Mrh- Fim- mutant of E. coli 536 and into a galE (rough) strain of Salmonella typhimurium yielded transformants that expressed both hemagglutination and fimbria production.


Woody encroachment slows decomposition and termite activity in an African savanna.
Woody encroachment can lead to a complete switch from open habitats to dense thickets, and has the potential to greatly alter the biodiversity and ecological functioning of grassy ecosystems across the globe. Plant litter decomposition is a critical ecosystem process fundamental to nutrient cycling and global carbon dynamics, yet little is known about how woody encroachment might alter this process. We compared grass decay rates of heavily encroached areas with adjacent nonencroached open areas in a semi-arid South African savanna using litterbags that allowed or excluded invertebrates. We also assessed the effect of woody encroachment on the activity of termites- dominant decomposer organisms in savanna systems. We found a significant reduction in decomposition rates within encroached areas, with litter taking twice as long to decay compared with open savanna areas. Moreover, invertebrates were more influential on grass decomposition in open areas and termite activity was substantially lower in encroached areas, particularly during the dry season when activity levels were reduced to almost zero. Our results suggest that woody encroachment created an unfavourable environment for invertebrates, and termites in particular, leading to decreased decomposition rates in these areas. We provide the first quantification of woody encroachment altering the functioning of African savanna ecosystems through the slowing of aboveground plant decomposition. Woody encroachment is intensifying across the globe, and our results suggest that substantial changes to the carbon balance and biodiversity of grassy biomes could occur.


Class I phosphoinositide 3-kinase p110beta is required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by macrophages.
Phosphoinositide 3-kinases (PI3Ks) play an important role in a variety of cellular functions, including phagocytosis. PI3Ks are activated during phagocytosis induced by several receptors and have been shown to be required for phagocytosis through the use of inhibitors such as wortmannin and LY294002. Mammalian cells have multiple isoforms of PI3K, and the role of the individual isoforms during phagocytosis has not been addressed. The class I PI3Ks consist of a catalytic p110 isoform associated with a regulatory subunit. Mammals have three genes for the class IA p110 subunits encoding p110alpha, p110beta, and p110delta and one gene for the class IB p110 subunit encoding p110gamma. Here we report a specific recruitment of p110beta and p110delta (but not p110alpha) isoforms to the nascent phagosome during apoptotic cell phagocytosis by fibroblasts. By microinjecting inhibitory antibodies specific to class IA p110 subunits, we have shown that p110beta is the major isoform required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by primary mouse macrophages. Macrophages from mice expressing a catalytically inactive form of p110delta showed no defect in the phagocytosis of apoptotic cells and IgG-opsonized particles, confirming the lack of a major role for p110delta in this process. Similarly, p110gamma-deficient macrophages phagocytosed apoptotic cells normally. Our findings demonstrate that p110beta is the major class I catalytic isoform required for apoptotic cell and Fcgamma receptor-mediated phagocytosis by primary macrophages.


Up-to-date management of carcinoma of the penis.
Squamous cell carcinoma is the most common tumour of the penis. It is a rare disease in Western countries, and it is often associated with phimosis, poor hygiene or human papilloma virus infection. It could be prevented or diagnosed early in most cases, but, due to cultural and educational reasons, it is often diagnosed late. Nodal metastases are relatively common, but distant dissemination is very rare. Radical surgery gives the best control of the primary tumour, but it is mutilating. Laser surgery for limited superficial lesions and sophisticated radiotherapy for relatively small infiltrating tumours have been successfully employed, alone or in combination with chemotherapy. The use of radical surgery can therefore be restricted to cases which are unsuitable for conservative treatment or to relapses. Survival mainly depends on nodal metastases, but management of regional lymph nodes is controversial. Radical inguinal or ileoinguinal lymphadenectomy can cure approximately 40-50% of patients with positive nodes, but nearly half of the patients with clinically enlarged nodes actually have no metastases. Invalidating oedema is a frequent complication of inguinal lymphadenectomy. The point is to restrict the operation to patients with positive nodes. Expectant policy can be dangerous because results of delayed lymphadenectomy are usually poor. Fine needle aspiration biopsy and imaging may be of help in diagnosing nodal metastases. Modified surgical procedures have been advocated in order to obtain a pathological staging of the inguinal nodes avoiding invalidating sequelae, but results are controversial. Depth of invasion, tumour grade and growth pattern are of help in identifying patients at a very high risk of harbouring nodal metastases. Squamous cell carcinoma of the penis is relatively responsive to chemotherapy. Limited experiences suggest that adjuvant chemotherapy can improve the long-term survival of patients with radically resected positive nodes, and primary chemotherapy can make resectable approximately 50% of cases with fixed inguinal metastases. However, chemotherapy alone is not curative for metastatic disease.


Skin Intrinsic Fluorescence and Selected Measures of Visual Function and aging in Older Adults.
Purpose: Functional, structural and metabolic decline in many systems and in combination contribute to biologic aging and may be manifest as increased risk of morbid events such as neuropathy, albuminuria, and coronary artery disease or mortality. A biologic marker of aging may be a useful tool in identifying persons at increased risk of morbidity or mortality. We have measured skin intrinsic fluorescence (SIF) in a group of older adults to determine whether this easily determined measure could serve as such a biomarker. Methods: Survivors of a population based study of older adults in a moderate sized Midwestern town. Of the 1181 persons participating, 939 had measures of skin intrinsic fluorescence (SIF) and at least one functional or diagnostic characteristic at the most recent examination. Characteristics such as blood pressure, forced expiratory volume, vision, time to walk a standard course and medical history and their associations with SIF measures were examined. Mortality after the last examination with respect to SIF was also investigated. There were 118 deaths among those who participated in this phase of the study. All analyses pertinent to these findings were adjusted for age. Results: SIF measures were significantly associated with low contrast sensitivity, more errors on frequency doubling technology testing (loss of peripheral vision), self-reported poor vision, slow gait, poor forced expiratory volume, and self-reported poor health. SIF was also associated with increased risk of death. All of these analyses were adjusted for age. Conclusions: Skin intrinsic fluorescence provides easily obtained markers of age-related functional outcomes, suggesting SIF measurements may be useful to identify persons who may benefit from more frequent medical scrutiny to decrease morbidity and mortality.


Endonuclease IV (nfo) mutant of Escherichia coli.
A cloned gene, designated nfo, caused overproduction of an EDTA-resistant endonuclease specific for apurinic-apyrimidinic sites in DNA. The sedimentation coefficient of the enzyme was similar to that of endonuclease IV. An insertion mutation was constructed in vitro and transferred from a plasmid to the Escherichia coli chromosome. nfo mutants had an increased sensitivity to the alkylating agents methyl methanesulfonate and mitomycin C and to the oxidants tert-butyl hydroperoxide and bleomycin. The nfo mutation enhanced the killing of xth (exonuclease III) mutants by methyl methanesulfonate, H2O2, tert-butyl hydroperoxide, and gamma rays, and it enhanced their mutability by methyl methanesulfonate. It also increased the temperature sensitivity of an xth dut (dUTPase) mutant that is defective in the repair of uracil-containing DNA. These results are consistent with earlier findings that endonuclease IV and exonuclease III both cleave DNA 5' to an apurinic-apyrimidinic site and that exonuclease III is more active. However, nfo mutants were more sensitive to tert-butyl hydroperoxide and to bleomycin than were xth mutants, suggesting that endonuclease IV might recognize some lesions that exonuclease III does not. The mutants displayed no marked increase in sensitivity to 254-nm UV radiation, and the addition of an nth (endonuclease III) mutation to nfo or nfo xth mutants did not significantly increase their sensitivity to any of the agents tested.


Specific B- and T-cell populations are associated with cognition in patients with epilepsy and antibody positive and negative suspected limbic encephalitis.
Neuropsychological impairments are major symptoms of autoimmune limbic encephalitis (LE) epilepsy patients. In LE epilepsy patients with an autoimmune response against intracellular antigens as well as in antibody-negative patients, the antibody findings and magnetic resonance imaging pathology correspond poorly to the clinical features. Here, we evaluated whether T- and B-cells are linked to cognitive impairment in these groups. In this cross-sectional, observational, case-controlled study, we evaluated 106 patients with adult-onset epilepsies with a suspected autoimmune etiology. We assessed verbal and visual memory, executive function, and mood in relation to the presence or absence of known auto-antibodies, and regarding T- and B-cell activity as indicated by flow cytometry (fluorescence-activated cell sorting = FACS, peripheral blood = PB and cerebrospinal fluid = CSF). 56% of the patients were antibody-negative. In the other patients, auto-antibodies were directed against intracellular antigens (GAD65, paraneoplastic: 38%), or cellular surface antigens (LGI1/CASPR2/NMDA-R: 6%). Excluding LGI1/CASPR2/NMDA-R, the groups with and without antibodies did not differ in disease features, cognition, or mood. CD4+ T-cells and CD8+ T-cells in blood and CD4+ T-cells in CSF were prominent in the auto-antibody positive group. Regression analyses indicated the role education, drug load, amygdala and/or hippocampal pathology, and CD4+ T-cells play in verbal memory and executive function. Depressed mood revealed no relation to flow cytometry results. Our results indicate a link between T- and B-cell activity and cognition in epilepsy patients with suspected limbic encephalitis, thus suggesting that flow cytometry results can provide an understanding of cognitive impairment in LE patients with autoantibodies against intracellular antigens.


Melanoma: the new perspectives from clinical and translational research.
The prognosis of metastatic melanoma has not changed throughout the 20th century. However, in the last decade, we have witnessed a continuous improvement in survival, with many long-term survivors. These results are largely because of the simultaneous development of the knowledge in the biology of metastatic malignant melanoma and of the relationship between the disease and the host's immune system that allowed the development of effective new treatments. In this overview, we summarize the therapies available today, their biological rationale, and the research field currently under investigation divided into three main chapters: target therapies, immunotherapies, and their combination.


PairsDB atlas of protein sequence space.
Sequence similarity/database searching is a cornerstone of molecular biology. PairsDB is a database intended to make exploring protein sequences and their similarity relationships quick and easy. Behind PairsDB is a comprehensive collection of protein sequences and BLAST and PSI-BLAST alignments between them. Instead of running BLAST or PSI-BLAST individually on each request, results are retrieved instantaneously from a database of pre-computed alignments. Filtering options allow you to find a set of sequences satisfying a set of criteria-for example, all human proteins with solved structure and without transmembrane segments. PairsDB is continually updated and covers all sequences in Uniprot. The data is stored in a MySQL relational database. Data files will be made available for download at ftp://nic.funet.fi/pub/sci/molbio. PairsDB can also be accessed interactively at http://pairsdb.csc.fi. PairsDB data is a valuable platform to build various downstream automated analysis pipelines. For example, the graph of all-against-all similarity relationships is the starting point for clustering protein families, delineating domains, improving alignment accuracy by consistency measures, and defining orthologous genes. Moreover, query-anchored stacked sequence alignments, profiles and consensus sequences are useful in studies of sequence conservation patterns for clues about possible functional sites.


The core histone-binding region of the murine cytomegalovirus 89K immediate early protein.
The gene regulatory immediate early protein, pp89, of murine cytomegalovirus interacts with both DNA-associated and isolated histones in vitro. We characterized the histone-binding region of pp89 and its cellular localization during cell division to examine the possible interaction between pp89 and chromatin. pp89 expressed constitutively in cell line BALB/c 3T3 IE1 does not interact with condensed chromatin. As observed in infected cells, pp89 is localized within the nucleus of cells during interphase but spreads throughout the cell plasma following degradation of the nuclear membrane during early mitosis. In late telophase, pp89 is reorganized within the nucleus. Analysis of pp89 deletion mutants and of fragments generated by cleavage at pH 2.5 revealed that the regions responsible for association with histone are located between amino acids 71 and 415, and are not identical with the domain that shows homology to histone H2B or the highly acidic carboxy-terminal region. A potential gene-activating role of the high affinity of pp89 for isolated histones and the low affinity for DNA-associated histones is discussed.


Hyposensitivity to amphetamine following exposure to chlorphenvinphos--protection by amphetamine preexposure.
We investigated the effect of an acute exposure to chlorphenvinphos (CVP), an organophosphate anticholinesterase, on amphetamine-induced open-field locomotion in rats. CVP was administered in a single i.p. dose of 1.0 mg/kg (1/10 of the LD50). All animals were challenged with 1.0 mg/kg amphetamine (AMPH) three weeks after the CVP exposure, i.e. after a time sufficient for acetylcholinesterase recovery. Some rats were also given AMPH three weeks before the CVP exposure. In rats challenged with AMPH only once after the CVP exposure, AMPH-induced open-field locomotion was significantly reduced. Such an effect was not observed in rats given AMPH three weeks before the CVP exposure. The results suggest that a single CVP exposure may result in persistent dopaminergic hyposensitivity, and that an amphetamine pretreatment may protect the rat against this effect.


Gamma radiation inactivation of coxsackievirus B-2.
The radioresistance of coxsackievirus B-2 was studied when the virus was suspended in Eagle minimal essential medium, distilled water, cooked ground beef, and raw ground beef and irradiated at various temperatures in a cobalt-60 gamma radiation source. The number of surviving viruses at given doses of radiation was determined by a plaque assay system. All destruction curves indicated a first-order reaction. When the virus was irradiated in minimal essential medium at temperatures of -30, -60, and -90 C, D values (in Mrad) were 0.69, 0.59, and 0.64, respectively. When the virus was suspended in water and irradiated at -90 C, the D value was 0.53. Cooked ground beef containing the virus was irradiated at temperatures ranging from 16 to -90 C. The D values were 0.70 (16 C), 0.76 (0.5 C), 0.68 (-30 C), 0.78 (-60 C), and 0.81 (-90 C). Raw ground beef containing the virus was irradiated at -30, -60, and -90 C, and the D values were respectively 0.75, 0.71, and 0.68. The D values indicate that the rate of viral inactivation was dependent on the suspending menstrum.


Validity review of performance measures.
A structured process for evaluating the validity of performance measures is essential if the rates generated by these measures are to be relied upon for large-scale programmes to improve clinical care. In this evaluation process a panel of objective clinicians who were not involved in developing the performance measures examines the criteria, coding instructions, and coding form. The panel's job is to determine the degree to which a sample of patient records containing treatment elements that have been flagged for nonconformance to certain criteria are truly nonconformant. Modification of the performance measure according to the recommendations of the panel provides an important means of reducing measurement error by eliminating obvious sources of false positive findings. This article describes the procedures followed and the materials used to validate the performance measures developed for the project to Develop and Evaluate Methods for Promoting Ambulatory Care Quality (DEMPAQ), a research project contracted by the Health Care Financing Administration of the United States government [1-3]. DEMPAQ is described in Fig. 1. In the DEMPAQ project, record review is conducted by entering data directly from patient ambulatory records into a computer system. Instructions for abstracting the data are displayed on the computer monitor.


Identification and influence of spatio-temporal outliers in urban air quality measurements.
Forty eight potential outliers in air pollution measurements taken simultaneously in Detroit, Michigan, USA and Windsor, Ontario, Canada in 2008 and 2009 were identified using four independent methods: box plots, variogram clouds, difference maps, and the Local Moran's I statistic. These methods were subsequently used in combination to reduce and select a final set of 13 outliers for nitrogen dioxide (NO2), volatile organic compounds (VOCs), total benzene, toluene, ethyl benzene, and xylene (BTEX), and particulate matter in two size fractions (PM2.5 and PM10). The selected outliers were excluded from the measurement datasets and used to revise air pollution models. In addition, a set of temporally-scaled air pollution models was generated using time series measurements from community air quality monitors, with and without the selected outliers. The influence of outlier exclusion on associations with asthma exacerbation rates aggregated at a postal zone scale in both cities was evaluated. Results demonstrate that the inclusion or exclusion of outliers influences the strength of observed associations between intraurban air quality and asthma exacerbation in both cities. The box plot, variogram cloud, and difference map methods largely determined the final list of outliers, due to the high degree of conformity among their results. The Moran's I approach was not useful for outlier identification in the datasets studied. Removing outliers changed the spatial distribution of modeled concentration values and derivative exposure estimates averaged over postal zones. Overall, associations between air pollution and acute asthma exacerbation rates were weaker with outliers removed, but improved with the addition of temporal information. Decreases in statistically significant associations between air pollution and asthma resulted, in part, from smaller pollutant concentration ranges used for linear regression. Nevertheless, the practice of identifying outliers through congruence among multiple methods strengthens confidence in the analysis of outlier presence and influence in environmental datasets.


The HIV-1 integrase mutant R263A/K264A is 2-fold defective for TRN-SR2 binding and viral nuclear import.
Transportin-SR2 (Tnpo3, TRN-SR2), a human karyopherin encoded by the TNPO3 gene, has been identified as a cellular cofactor of HIV-1 replication, specifically interacting with HIV-1 integrase (IN). Whether this interaction mediates the nuclear import of HIV remains controversial. We previously characterized the TRN-SR2 binding interface in IN and introduced mutations at these positions to corroborate the biological relevance of the interaction. The pleiotropic nature of IN mutations complicated the interpretation. Indeed, all previously tested IN interaction mutants also affected RT. Here we report on a virus with a pair of IN mutations, IN(R263A/K264A), that significantly reduce interaction with TRN-SR2. The virus retains wild-type reverse transcription activity but displays a block in nuclear import and integration, as measured by quantitative PCR. The defect in integration of this mutant resulted in a smaller increase in the number of two-long terminal repeat circles than for virus specifically blocked at integration by raltegravir or catalytic site mutations (IN(D64N/D116N/E152Q)). Finally, using an eGFP-IN-labeled HIV fluorescence-based import assay, the defect in nuclear import was corroborated. These data altogether underscore the importance of the HIV-IN TRN-SR2 protein-protein interaction for HIV nuclear import and validate the IN/TRN-SR2 interaction interface as a promising target for future antiviral therapy.


HLA-DR in Cytotoxic T Lymphocytes Predicts Breast Cancer Patients' Response to Neoadjuvant Chemotherapy.
Prediction of breast cancer response to Neoadjuvant Chemotherapy (NACT) is an urgent need to promptly direct non-responder patients to alternative therapies. Infiltrating T lymphocytes, namely cytotoxic T lymphocytes (CTLs) have been appointed as predictors of response. However, cancer cells have the ability to dampen CTLs' activity and thus, the prognostic value of the CTLs, per se, is debatable. Here, we disclose that more than the occurrence of CTLs, it is their activation state, revealed by HLA-DR expression, that can accurately predict response to NACT. Flow cytometry analysis of breast cancer biopsies showed that the frequency of CTLs and other lymphocytes were similar regardless disease stage and between NACT responders and non-responders. However, only breast cancer patients without axillary lymph node metastasis and NACT responders have HLA-DRhi CTLs. Interestingly, HLA-DR levels in tumor CTLs is correlated with HLA-DR levels in systemic CTLs. These HLA-DR+ CTLs produce IFN-γ and Granzyme B, enlightening their effector and probable anti-tumor activity profile. Moreover, the level of HLA-DR in CTLs is negatively correlated with the level of HLA-DR in T regulatory lymphocytes and with immunosuppressive and pro-tumor molecules in the tumor microenvironment. Hence, HLA-DR levels in CTLs is a highly sensitive and specific potential predictive factor of NACT-response, which can be assessed in blood to guide therapeutic decisions.


Meta analysis of risk factors for colorectal cancer.
To study the risk factors for colorectal cancer in China. A meta-analysis of the risk factors of colorectal cancer was conducted for 14 case-control studies, and reviewed 14 reports within 13 years which included 5034 cases and 5205 controls. Dersimonian and Laird random effective models were used to process the results. Meta analysis of the 14 studies demonstrated that proper physical activities and dietary fibers were protective factors (pooled OR<0.8), while fecal mucohemorrhage, chronic diarrhea and polyposis were highly associated with colorectal cancer (all pooled OR>4). The stratified results showed that different OR values of some factors were due to geographic factors or different resources. Risks of colorectal cancer are significantly associated with the histories of intestinal diseases or relative symptoms, high lipid diet, emotional trauma and family history of cancers. The suitable physical activities and dietary fibers are protective factors.


Perinatal death and respiratory apparatus dysgenesis due to a bis (dichloroacetyl) diamine.
N, N1-bis (dichloroacetyl) diamine 1, 8-octomethylenediamine (WIN 18,446) is an experimental drug which was first investigated as a male contraceptive. It is soluble in lipid solvents but not in water. The administration of 1,200 to 1,600 mg/kg to pregnant rats on the tenth day of gestation produced multiorgan fetal malformations. Smaller doses, 400 to 800 mg/kg, especially if divided over 2 or 3 days, caused perinatal death. Thus, 60 to 100% of offspring of rats given WIN 18,446 on the tenth and 11th days of gestation died at birth or within 4 days (Taleporos et al., 78). The present study investigated such deaths. At doses of 200 mg/kg on day 10 or 50 mg/kg on days 10 and 11, 67% of offspring had defective or absent diaphragms, 48% had tracheobronchiomegaly with cystic lungs, and 67% had pleural hemorrhage. At doses of 100 mg/kg given on 1 day or 25 mg/kg each day for 2 days, 50% had tracheobronchiomegaly with cystic lungs and rudimentary acini. At lower doses (18.8 mg/kg X 2 or 12.4 mg/kg X 3), a majority of fetal lungs had rudimentary acini, thick septa, few capillaries, and wide cuffs of perivascular connective tissue. Thus, a chemical given during organogenesis produced dysgenesis of the respiratory apparatus. Varying the dose produced malformed lungs with persistently deficient acini which model such human lung faults as tracheobronchiomegaly (Mournier-Kuhn Syndrome; Mounier-Kuhn, '32), bronchiolar dysplasia (Wilson-Mikity Syndrome), and perinatal death with acinar failure resembling neonatal hyaline membrane disease.


Blood transfusion is associated with increased risk of perioperative complications and prolonged hospital duration of stay among patients undergoing amputation.
We evaluated the outcomes of patients undergoing major lower-extremity amputation who received packed red blood cell transfusion. Using the dataset of the National Surgical Quality Improvement Program (2005-2011), we examined 5,739 above-knee and 6,725 below-knee amputations. Patients were stratified by perioperative (preoperative, intraoperative, or postoperative) blood transfusion. Outcomes included perioperative mortality, myocardial infarction (MI), thromboembolism, and duration of stay (DOS) at the hospital. Adjusted comparisons of outcomes between transfused and not-transfused patients were performed by matching the 2 groups for age, smoking, diabetes, renal failure, coronary artery disease, classification of the American Society of Anesthesiologists, functional status, and procedure type. Multivariable logistic and gamma regression were used to examine associations between transfusion and outcomes. Of the 12,464 amputations in the study cohort 2,133 (17%) required transfusion. The majority of the cases were performed for critical limb ischemia (8,205 amputations; 66%) and the overall 30-days mortality was 9%. In both crude and matched cohorts, although perioperative mortality and cardiac complication rates were similar, transfusion was associated with a greater incidence of pneumonia (crude: 6.1% vs 3%, P < .001; matched: 5.9% vs 3.7%, P < .001), thromboembolism (2.5% vs 1.6%, P = .003; 2.5% vs 1.4%, P = .002) and longer DOS (18 ± 19 vs 13.6 ± 14.3 days, P < .001; 17.8 ± 18.4 vs 14.2 ± 14.5 days, P < .001). Multivariable adjustment for confounding variables in the crude cohort demonstrated that transfusion was independently associated with a greater odds of perioperative pneumonia (odds ratio [OR]:1.6; 95% confidence interval [CI]:1.3-2; P < .001), thromboembolism (OR 1.3, 95% CI 1.0-1.9, P = .09) and longer DOS (mean ratio: 1.1; 95% CI 1.1-1.6; P = .006). Among patients who had major lower-extremity amputation, perioperative transfusion independently predicted greater risks for perioperative pneumonia, thromboembolism, and prolonged hospital DOS.


Increasing minimum daily temperatures are associated with enhanced pesticide use in cultivated soybean along a latitudinal gradient in the mid-western United States.
Assessments of climate change and food security often do not consider changes to crop production as a function of altered pest pressures. Evaluation of potential changes may be difficult, in part, because management practices are routinely utilized in situ to minimize pest injury. If so, then such practices, should, in theory, also change with climate, although this has never been quantified. Chemical (pesticide) applications remain the primary means of managing pests in industrialized countries. While a wide range of climate variables can influence chemical use, minimum daily temperature (lowest 24 h recorded temperature in a given year) can be associated with the distribution and thermal survival of many agricultural pests in temperate regions. The current study quantifies average pesticide applications since 1999 for commercial soybean grown over a 2100 km North-South latitudinal transect for seven states that varied in minimum daily temperature (1999-2013) from -28.6°C (Minnesota) to -5.1°C (Louisiana). Although soybean yields (per hectare) did not vary by state, total pesticide applications (kg of active ingredient, ai, per hectare) increased from 4.3 to 6.5 over this temperature range. Significant correlations were observed between minimum daily temperatures and kg of ai for all pesticide classes. This suggested that minimum daily temperature could serve as a proxy for pesticide application. Longer term temperature data (1977-2013) indicated greater relative increases in minimum daily temperatures for northern relative to southern states. Using these longer-term trends to determine short-term projections of pesticide use (to 2023) showed a greater comparative increase in herbicide use for soybean in northern; but a greater increase in insecticide and fungicide use for southern states in a warmer climate. Overall, these data suggest that increases in pesticide application rates may be a means to maintain soybean production in response to rising minimum daily temperatures and potential increases in pest pressures.


Description of patients undergoing high risk angioplasty. Analyses of various databases.
As the pharmacological and mechanical management of angioplasty procedures advances, the description of risk incurred during and after the procedure becomes increasingly important. A variety of measures from different sorts of databases have been devised to facilitate this task. Large public databases generally include descriptors of overall physical health and are most useful in predicting catastrophic outcomes, such as mortality. Data from smaller angiographic databases are more useful in providing specific descriptors of more common but less catastrophic events, such as abrupt closure, myocardial infarction, and emergency coronary artery bypass. Most complications are associated with abrupt closure of the vessel dilated. In modern PTCA series, the risk of serious ishcaemic complications ranges from as low as 3% to as high as 12.9%. Since the inception of angioplasty, it has been observed repeatedly that angiographic characteristics of target lesions can predict the likelihood of both success and ischaemic complications. It has also become apparent that such clinical factors as unstable angina with ST segment changes or recent myocardial infarction also play important roles by providing a thrombogenic milieu in which the risk of intravascular thrombosis is increased. Although advances in operator skills and equipment, as well as new devices have dramatically increased the degree of lesion complexity which may be approached with PTCA, the actual characteristics which are associated with abrupt closure have undergone little change.


Disability, gender, and employment relationships in Africa: The case of Ghana.
The exploratory quantitative study sought to develop an understanding about the relationships among disability, gender and employment in Northern Ghana. A total of 110 individuals with disabilities (20-60 years) from various disability groups participated in the study. The results indicate that many persons with disabilities are unemployed, the majority being women. Discrimination is cited as the greatest barrier to the employment of persons with disabilities, particularly women. The majority of persons with disabilities, typically women, live in poverty; given that some are unemployed and those who are employed worked mostly in marginal, seasonal and menial jobs. Persons with disabilities also experience several challenges on the job, including negative perceptions about their capabilities, discrimination and exclusion, irrespective of the employment sector and disability type. Educational interventions such as workshops, documenting and showcasing success stories of persons with disabilities could be helpful to reduce negative perceptions about their capabilities as well as discrimination against them. Government intervention to support persons with disabilities with start-up capital and funding for formal education is also recommended as these two elements were identified respectively as barriers to self-employment and employment in the public/private sectors. Government interventions to create educational opportunities for persons with disabilities are essential given that lower educational attainment affect their employment.


Enrollment in prescription drug insurance: the interaction of numeracy and choice set size.
To determine how choice set size affects decision quality among individuals of different levels of numeracy choosing prescription drug plans. Members of an Internet-enabled panel age 65 and over were randomly assigned to sets of prescription drug plans varying in size from 2 to 16 plans from which they made a hypothetical choice. They answered questions about enrollment likelihood and the costs and benefits of their choice. The measure of decision quality was enrollment likelihood among those for whom enrollment was beneficial. Enrollment likelihood by numeracy and choice set size was calculated. A model of moderated mediation was analyzed to understand the role of numeracy as a moderator of the relationship between the number of plans and the quality of the enrollment decision and the roles of the costs and benefits in mediating that relationship. More numerate adults made better decisions than less numerate adults when choosing among a small number of alternatives but not when choice sets were larger. Choice set size had little effect on decision making of less numerate adults. Differences in decision making costs between more and less numerate adults helped explain the effect of choice set size on decision quality. Interventions to improve decision making in the context of Medicare Part D may differentially affect lower and higher numeracy adults. The conflicting results on choice overload in the psychology literature may be explained in part by differences amongst individuals in how they respond to choice set size.


Specific inter-domain interactions stabilize a compact HIV-1 Gag conformation.
HIV-1 Gag is a large multidomain poly-protein with flexible unstructured linkers connecting its globular subdomains. It is compact when in solution but assumes an extended conformation when assembled within the immature HIV-1 virion. Here, we use molecular dynamics (MD) simulations to quantitatively characterize the intra-domain interactions of HIV-1 Gag. We find that the matrix (MA) domain and the C-terminal subdomain CActd of the CA capsid domain can form a bound state. The bound state, which is held together primarily by interactions between complementary charged and polar residues, stabilizes the compact state of HIV-1 Gag. We calculate the depth of the attractive free energy potential between the MA/ CActd sites and find it to be about three times larger than the dimerization interaction between the CActd domains. Sequence analysis shows high conservation within the newly-found intra-Gag MA/CActd binding site, as well as its spatial proximity to other well known elements of Gag -such as CActd's SP1 helix region, its inositol hexaphosphate (IP6) binding site and major homology region (MHR), as well as the MA trimerization site. Our results point to a high, but yet undetermined, functional significance of the intra-Gag binding site. Recent biophysical experiments that address the binding specificity of Gag are interpreted in the context of the MA/CActd bound state, suggesting an important role in selective packaging of genomic RNA by Gag.


Effects of bile salts and prostaglandins on sodium transport in isolated rat gastric mucosa.
To determine whether prostaglandins may protect against bile salt inhibition of ion transport in the stomach, gastric mucosal tissue was isolated from the rat and mounted in flux chambers. Transport of Na+ was traced with radioisotopes in the absence of bile salts and then in the presence of conjugated taurocholate or unconjugated deoxycholate at low, intermediate and high mucosal concentrations (1, 5 and 15 mmol/1). At a high (7.40) or low (3.4) mucosal pH, only the unconjugated deoxycholate inhibited active Na+ transport from mucosa to submucosa with respect to untreated controls. Inhibition of Na+ transport was apparent at a low level of deoxycholate, which also inhibited the electrical potential difference. Intermediate and high levels of deoxycholate lowered the tissue resistance. When the tissues were exposed to mucosal prostaglandin E2 or its 16,16-dimethyl analogue before and during acidified taurocholate administration, Na+ transport was not changed significantly but the electrical resistance remained high. Thus, unconjugated bile salt is more potent than conjugated bile salt in inhibiting Na+ transport and breaking the gastric mucosal barrier, and prostaglandins may afford some small protection.


Spectral reflectance from a soybean canopy exposed to elevated CO2 and O3.
By affecting the physiology and structure of plant canopies, increasing atmospheric CO(2) and O(3) influence the capacity of agroecosystems to capture light and convert that light energy into biomass, ultimately affecting productivity and yield. The objective of this study was to determine if established remote sensing indices could detect the direct and interactive effects of elevated CO(2) and elevated O(3) on the leaf area, chlorophyll content, and photosynthetic capacity of a soybean canopy growing under field conditions. Large plots of soybean (Glycine max) were exposed to ambient air (∼380 μmol CO(2) mol(-1)), elevated CO(2) (∼550 μmol mol(-1)), elevated O(3) (1.2× ambient), and combined elevated CO(2) plus elevated O(3) at the soybean free air gas concentration enrichment (SoyFACE) experiment. Canopy reflectance was measured weekly and the following indices were calculated from reflectance data: near infrared/red (NIR/red), normalized difference vegetation index (NDVI), canopy chlorophyll content index (chl. index), and photochemical reflectance index (PRI). Leaf area index (LAI) also was measured weekly. NIR/red and LAI were linearly correlated throughout the growing season; however, NDVI and LAI were correlated only up to LAI values of ∼3. Season-wide analysis demonstrated that elevated CO(2) significantly increased NIR/red, PRI, and chl. index, indicating a stimulation of LAI and photosynthetic carbon assimilation, as well as delayed senescence; however, analysis of individual dates resolved fewer statistically significant effects of elevated CO(2). Exposure to elevated O(3) decreased LAI throughout the growing season. Although NIR/red showed the same trend, the effect of O(3) on NIR/red was not statistically significant. Season-wide analysis showed significant effects of O(3) on PRI; however, analysis of individual dates revealed that this effect was only statistically significant on two dates. Elevated O(3) had minimal effects on the total canopy chlorophyll index. PRI appeared to be more sensitive to decreased photosynthetic capacity of the canopy as a whole compared with previously published single leaf gas exchange measurements at SoyFACE, possibly because PRI integrates the reflectance signal of older leaves with accumulated O(3) damage and healthy young, upper canopy leaves, enabling detection of significant decreases in photosynthetic carbon assimilation which have not been detected in previous studies which measured gas exchange of upper canopy leaves. When the canopy was exposed to elevated CO(2) and O(3) simultaneously, the deleterious effects of elevated O(3) were diminished. Reflectance data, while less sensitive than direct measurements of physiological/structural parameters, corroborate direct measurements of LAI and photosynthetic gas exchange made during the same season, as well as results from previous years at SoyFACE, demonstrating that these indices accurately represent structural and physiological effects of changing tropospheric chemistry on soybean growing in a field setting.


Multivariate inference of pathway activity in host immunity and response to therapeutics.
Developing a quantitative view of how biological pathways are regulated in response to environmental factors is central for understanding of disease phenotypes. We present a computational framework, named Multivariate Inference of Pathway Activity (MIPA), which quantifies degree of activity induced in a biological pathway by computing five distinct measures from transcriptomic profiles of its member genes. Statistical significance of inferred activity is examined using multiple independent self-contained tests followed by a competitive analysis. The method incorporates a new algorithm to identify a subset of genes that may regulate the extent of activity induced in a pathway. We present an in-depth evaluation of specificity, robustness, and reproducibility of our method. We benchmarked MIPA's false positive rate at less than 1%. Using transcriptomic profiles representing distinct physiological and disease states, we illustrate applicability of our method in (i) identifying gene-gene interactions in autophagy-dependent response to Salmonella infection, (ii) uncovering gene-environment interactions in host response to bacterial and viral pathogens and (iii) identifying driver genes and processes that contribute to wound healing and response to anti-TNFα therapy. We provide relevant experimental validation that corroborates the accuracy and advantage of our method.


Sleep nasendoscopy: a technique of assessment in snoring and obstructive sleep apnoea.
It appears that uvulopalatopharyngoplasty (UVPP) is a reliable procedure for reducing snoring, but much less reliable when used as a treatment for OSAS. This is thought to be because of poor patient selection in that the site of the problem is not always the site of the operation. We present the technique of sleep nasendoscopy which allows direct visualization of the site or sites of obstruction in a sleeping patient. Our study has shown that there are patients with obstructive sleep apnoea syndrome (OSAS) in whom the only site of pharyngeal obstruction is at the velopharynx. These patients should do well with the relatively simple procedure of UVPP. This is not true for many other OSAS patients in whom we found that obstruction was multisegmental. This helps to explain the frequently poor results of UVPP in OSAS patients. We feel that this form of preoperative assessment will avoid unnecessary surgery.


Alpha 2-plasmin inhibitor-plasmin complexes in synovial fluid.
The role of the fibrinolytic enzyme system in inflammatory joint disease has been addressed by several experimental approaches. In our study we examined synovial fluid (SF) in several different diseases for alpha 2-plasmin inhibitor-plasmin complexes by an enzyme linked immunosorbent assay (ELISA). These complexes are present in both rheumatoid and nonrheumatoid SF, with highest levels seen in septic arthritis. Thus the plasminogen system is intact in inflammatory SF, and plasmin may play a role in the local inflammatory process.


Ultrasensitive carbon nanotube-based biosensors using antibody-binding fragments.
We report a method to build ultrasensitive carbon nanotube-based biosensors using immune binding reaction. Here carbon nanotube-field effect transistors (CNT-FETs) were functionalized with antibody-binding fragments as a receptor, and the binding event of target immunoglobulin G (IgG) onto the fragments was detected by monitoring the gating effect caused by the charges of the target IgG. Because the biosensors were used in buffer solution, it was crucial to use small-size receptors so that the charged target IgG could approach the CNT surface within the Debye length distance to give a large gating effect. The results show that CNT-FET biosensors using whole antibody had very low sensitivity (detection limit approximately 1000 ng/ml), whereas those based on small Fab fragments could detect 1 pg/ml (approximately 7 fM level). Moreover, our Fab-modified CNT-FET could successfully block the nontarget proteins and could selectively detect the target protein in an environment similar to that of human serum electrolyte. Significantly, this strategy can be applied to general antibody-based detection schemes, and it should enable the production of label-free ultrasensitive electronic biosensors to detect clinically important biomarkers for disease diagnosis.


[Improvement of learning and memory functions by GABAB receptor antagonists in mice].
In one trial passive avoidance response in mice, the effects of gamma-aminobutyric acid (GABA)B receptor agonist baclofen and antagonist CGP35348 and CGP36742 on acquisition, consolidation and retrieval of memory were observed. The results showed that the antagonists could significantly promote the acquisition impairment induced by baclofen, the consolidation impairment induced by baclofen and NaNO2, and the retrieval impairment induced by baclofen and 30% alcohol. These results suggest that the GABAB receptor antagonists may become a novel type of drug for the treatment of Alzheimer's disease.


Interventions for replacing missing teeth: treatment of perimplantitis.
One of the key factors for the long-term success of oral implants is the maintenance of healthy tissues around them. Bacterial plaque accumulation induces inflammatory changes in the soft tissues surrounding oral implants and it may lead to their progressive destruction (perimplantitis) and ultimately to implant failure. Different treatment strategies for perimplantitis have been suggested, however it is unclear which are the most effective. To identify the most effective interventions for treating perimplantitis around osseointegrated dental implants. We searched the Cochrane Oral Health Group's Trials Register, CENTRAL, MEDLINE and EMBASE. Handsearching included several dental journals. We checked the bibliographies of the identified randomised controlled trials (RCTs) and relevant review articles for studies outside the handsearched journals. We wrote to authors of all identified RCTs, to more than 55 dental implant manufacturers and an Internet discussion group to find unpublished or ongoing RCTs. No language restrictions were applied. The last electronic search was conducted on 7th January 2010. All RCTs comparing agents or interventions for treating perimplantitis around dental implants. Screening of eligible studies, assessment of the methodological quality of the trials and data extraction were conducted in duplicate and independently by two review authors. We contacted the authors for missing information. Results were expressed as random-effects models using mean differences for continuous outcomes and risk ratios for dichotomous outcomes with 95% confidence intervals (CI). Heterogeneity was to be investigated including both clinical and methodological factors. Twelve eligible trials were identified, but five were excluded. The following procedures were tested: (1) use of local antibiotics versus ultrasonic debridement; (2) benefits of adjunctive local antibiotics to debridement; (3) different techniques of subgingival debridement; (4) laser versus manual debridement and chlorhexidine irrigation/gel; (5) systemic antibiotics plus resective surgery plus two different local antibiotics with and without implant surface smoothening; and (6) nanocrystalline hydroxyapatite versus Bio-Oss and resorbable barriers. Follow-up ranged from 3 months to 4 years. The only statistically significant differences were observed in two trials judged to be at high risk of bias. After 4 months, adjunctive local antibiotics to manual debridement in patients who lost at least 50% of the bone around implants showed improved mean probing attachment levels (PAL) of 0.61 mm and reduced probing pockets depths (PPD) of 0.59 mm. After 4 years, patients with perimplant infrabony defects > 3 mm treated with Bio-Oss and resorbable barriers gained 1.4 mm more PAL and PPD than patients treated with a nanocrystalline hydroxyapatite. There is very little reliable evidence suggesting which could be the most effective interventions for treating perimplantitis. This is not to say that currently used interventions are not effective. The use of local antibiotics in addition to manual subgingival debridement was associated with a 0.6 mm additional improvement for PAL and PPD over a 4-month period in patients affected by severe forms of perimplantitis. After 4 years, improved PAL and PPD of about 1.4 mm were obtained when using Bio-Oss with resorbable barriers compared to a nanocrystalline hydroxyapatite in perimplant infrabony defects. In four trials, the control therapy which basically consisted of a simple subgingival mechanical debridement seemed to be sufficient to achieve similar results to the more complex and expensive therapies. Follow-up longer than 1 year suggested recurrence of perimplantitis up to 100% of the treated cases for some of the tested interventions. Sample sizes were very small and follow-up too short, therefore these findings have to be considered with great caution. Larger well-designed RCTs with follow-up longer than 1 year are needed.


Hemiarthroplasty Versus Total Shoulder Arthroplasty for Shoulder Osteoarthritis: A Matched Comparison of Return to Sports.
Return to activity is a commonly used indication for shoulder hemiarthroplasty (HA) compared with total shoulder arthroplasty (TSA). Despite clinical studies demonstrating better functional outcomes after TSA, the literature has failed to show a difference in return to sports. To compare rates of return to sports in a matched cohort of TSA and HA patients with a preoperative diagnosis of glenohumeral osteoarthritis (OA). Cohort study; Level of evidence, 3. A prospectively collected registry was queried retrospectively for consecutive patients who underwent HA. Inclusion criteria were preoperative diagnosis of OA and more than 2 years of follow-up. After meeting the inclusion criteria, all HA patients were statistically matched to a TSA patient. All patients had end-stage OA with significant glenohumeral joint space narrowing. At final follow-up, 40 HA patients and 40 TSA patients were available. The average (±SD) age at surgery was 65.7 ± 10.5 years and 66.2 ± 9.6 years for the HA and TSA groups, respectively (P = .06). Average follow-up was 62.0 months and 61.1 months for the HA and TSA groups, respectively (P = .52). Average American Shoulder and Elbow Surgeons scores improved from 36.3 to 70.2 for HA patients and from 34.0 to 78.5 for TSA patients (P < .001 for both); final scores were not significantly different between groups (P = .21). Average visual analog scale pain scores improved from 6.3 to 2.2 for HA patients and from 6.1 to 0.6 for TSA patients (P < .001 for both). HA patients had significantly worse final visual analog scale scores compared with the TSA group (P = .002). Significantly more TSA patients were satisfied with their surgery compared with HA patients (100% vs 70%) (P = .01). Of both groups, 65.5% of HA patients (19 of 29) returned to at least 1 sport postoperatively compared with 97.3% of TSA patients (36 of 37) (P < .001). Average timing for return to full sports was 5.5 ± 4.2 months and 5.4 ± 3.1 months for the HA and TSA groups, respectively (P = .92). Significantly more TSA patients returned to higher upper extremity use sports (P = .01). In patients with OA, rate of return to sports was significantly better after TSA compared with HA. HA patients had significantly more pain, worse surgical satisfaction, and decreased ability to return to high upper extremity use sports. For patients with OA who wish to return to sporting activities, these results help manage expectations.


Genetic mapping of three quantitative trait loci for soybean aphid resistance in PI 567324.
Host-plant resistance is an effective method for controlling soybean aphid (Aphis glycines Matsumura), the most damaging insect pest of soybean (Glycine max (L.) Merr.) in North America. Recently, resistant soybean lines have been discovered and at least four aphid resistance genes (Rag1, Rag2, Rag3 and rag4) have been mapped on different soybean chromosomes. However, the evolution of new soybean aphid biotypes capable of defeating host-plant resistance conferred by most single genes demonstrates the need for finding germplasm with multigenic resistance to the aphid. This study was conducted to map quantitative trait loci (QTL) for aphid resistance in PI 567324. We identified two major QTL (QTL_13_1 and QTL_13_2) for aphid resistance on soybean chromosome 13 using 184 recombinant inbred lines from a 'Wyandot' × PI 567324 cross. QTL_13_1 was located close to the previously reported Rag2 gene locus, and QTL_13_2 was close to the rag4 locus. A minor QTL (QTL_6_1) was also detected on chromosome 6, where no gene for soybean aphid resistance has been reported so far. These results indicate that PI 567324 possesses oligogenic resistance to the soybean aphid. The molecular markers closely linked to the QTL reported here will be useful for development of cultivars with oligogenic resistance that are expected to provide broader and more durable resistance against soybean aphids compared with cultivars with monogenic resistance.


Structural studies of the O-antigen polysaccharide from Escherichia coli O138.
The structure of the O-antigen polysaccharide from Escherichia coli O138 has been determined. NMR spectroscopy, together with component and methylation analyses, of native and reduced polysaccharide were the principal methods used. The sequence of the sugar residues could be determined by NOESY and heteronuclear multiple bond connectivity (HMBC) NMR experiments. It is concluded that the polysaccharide is composed of tetrasaccharide repeating units with the following structure: [structure: see text].


Hippocampal CA1 lacunosum-moleculare interneurons: modulation of monosynaptic GABAergic IPSCs by presynaptic GABAB receptors.
1. Whole cell patch-clamp recordings were employed to characterize monosynaptic inhibitory postsynaptic currents (IPSCs) in morphologically and electrophysiologically identified interneurons located in the stratum lacunosum moleculare, or near the border of the stratum radiatum (LM interneurons), in the CA1 region of hippocampal slices taken from 3- to 4-wk-old rats. Monosynaptic IPSCs, evoked in the presence of glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM) and D-2-amino-5-phosphopentanoate (APV; 50 microM) were biphasic. The gamma-aminobutyric acid-A (GABAA) receptor antagonist, bicuculline (20 microM), blocked the fast IPSC, and the slow IPSC was blocked by the GABAB receptor antagonist CGP35348 (500 microM). 2. Monosynaptic IPSCs were evoked by electrical stimulation in several distant regions including the stratum radiatum, the stratum oriens, the stratum lacunosum-moleculare, and the molecular layer of dentate gyrus, suggesting an extensive network of inhibitory interneurons in the hippocampus. In paired recordings of CA1 interneurons and pyramidal cells, IPSCs were evoked by electrical stimulation of most of these distal regions with the exception of the molecular layer of dentate gyrus, which evoked an IPSC only in LM interneurons. 3. Frequent (> 0.1 Hz) stimulation depressed the evoked IPSCs. With a paired-pulse protocol, the second IPSC was depressed and the maximal depression (40-50%) was observed with an interstimulus interval of 100-200 ms. 4. The GABAB receptor agonist baclofen (1 microM) reduced the amplitude of evoked IPSCs and the paired-pulse depression of the second IPSC. The GABAB receptor antagonist CGP35348 (0.5-1 mM) had no significant effect on the amplitude of isolated IPSCs. However, CGP35348 reduced but did not fully block paired-pulse depression, suggesting that this depression is partly due to the activation of presynaptic GABAB receptors. 5. The paired-pulse depression depended on the level of transmitter release. Potentiation of synaptic release of GABA, by increasing the extracellular Ca2+ concentration to 4 mM and reducing the extracellular Mg2+ concentration to 0.1 mM, enhanced the depression. Reduction of transmitter release by increasing extracellular Mg2+ concentration to 7 mM diminished the paired-pulse depression of IPSCs. After potentiation of transmitter release, CGP35348 was less efficient in reducing the paired-pulse depression, suggesting that enhancement of depression by high-calcium/low-magnesium medium was preferentially due to the potentiation of a GABAB-independent component. 6. In summary, monosynaptic IPSCs recorded in LM interneurons show similar features to those recorded in pyramidal cells. The strong correlation between the level of transmitter release and the degree of paired-pulse depression may have important physiological consequences, because in synapses with a high level of activity and a high level of GABA release, inhibition is powerful, but depression can develop more readily.


Development and pharmacology of angiotensin converting enzyme inhibitors.
Angiotensin converting enzyme inhibitors represent a new class of agents which were designed to retain only that unique property. Because of this inhibition, administration of these agents results in the significant reduction of elevated blood pressure of various etiologies and in the amelioration of symptoms associated with congestive heart failure resistant to digitalis glycosides and diuretics. The mechanism(s) of action is not entirely certain but almost certainly resides in the inhibition of ACE. Certainly, the future will see still further advances made in this therapeutic area as well as with other inhibitors of the renin-angiotensin system.


Genomic organization of two novel genes on human Xq28: compact head to head arrangement of IDH gamma and TRAP delta is conserved in rat and mouse.
In this paper we present the entire genomic sequence as well as the cDNA sequence of two new human genes encoding the gamma subunit of the NAD(+)-dependent isocitrate dehydrogenase (H-IDH gamma) and the translocon-associated protein delta subunit (TRAP delta). These genes are located on region q28 of the human X chromosome, approximately 70 kb telomeric to the adrenoleukodystrophy locus (ALD). The sequences of the transcripts of both genes were obtained by searching the EST database with genomic data. Identified ESTs were completely sequenced and assembled to cDNAs comprising the entire coding region. For IDH gamma, several EST clones indicate differential splicing. IDH gamma and TRAP delta are arranged in a compact head to head manner. The nontranscribed intergenic region represents only 133 bp and is embedded in a CpG island. The CpG island obviously functions as a bidirectional promoter to initiate the transcription of both functionally unrelated genes with quite distinct expression patterns. This exceptional gene arrangement prompted us to clone and sequence genomic DNA fragments containing the homologous intergenic regions of rat and mouse. We show that in both species this area is similarly compact and represents less than 249 bp in rat and not more than 164 bp in mouse. In both cases this intergenic region is embedded in a CpG island and is highly conserved with nucleotide identity values ranging from 70.1% between human and rat to 92.6% between mouse and rat.


Diet and urine estrogens among postmenopausal women.
Creatinine-adjusted levels of estrone, estradiol and estriol were determined in overnight urine specimens from 88 postmenopausal women from Athens, Greece, and were correlated with daily nutrient intakes estimated through a semiquantitative food frequency questionnaire. Although obesity was positively and significantly related to all three urinary estrogens and their total, none of the investigated macro- or micronutrients was significantly or suggestively associated to any of these urinary estrogens, after controlling for energy intake, reproductive and biosocial variables. These results suggest that quantitative rather than qualitative aspects of nutrition affect the levels of postmenopausal estrogens, although endogenous factors could also be responsible for the association of these estrogens with obesity. Alcohol intake was also positively associated with urinary estrogens (mainly estrone and estradiol), after controlling for energy intake, obesity and the other indicated variables.


Semantic Interoperability, E-Health and Australian Health Statistics.
E-health implementation in Australia will depend upon interoperable computer systems to share information and data across the health sector. Semantic interoperability, which preserves the meaning of information and data when it is shared or re-purposed, is critical for safe clinical care, and also for any re-use of the information or data for other purposes. One such re-use is for national health statistics. Usable statistics rely on comparable and consistent data, and current practice is to use agreed national data standards to achieve this. The standardisation and interoperability needed to support e-health should also provide strong support for national health statistics. This report discusses some of the semantic interoperability issues involved in moving from the current data supply process for national health statistics to an e-health-enabled future.


Novel brominated flame retardants: a review of their analysis, environmental fate and behaviour.
This review summarises current knowledge about production volumes, physico-chemical properties, analysis, environmental occurrence, fate and behaviour and human exposure to the "novel" brominated flame retardants (NBFRs). We define the term NBFRs as relating to BFRs which are new to the market or newly/recently observed in the environment. Restrictions and bans on the use of some polybrominated diphenyl ether (PBDE) formulations, in many jurisdictions, have created a market for the use of NBFRs. To date, most data on NBFRs have arisen as additional information generated by research designed principally to study more "traditional" BFRs, such as PBDEs. This has led to a wide variety of analytical approaches for sample extraction, extract purification and instrumental analysis of NBFRs. An overview of environmental occurrence in abiotic matrices, aquatic biota, terrestrial biota and birds is presented. Evidence concerning the metabolism and absorption of different NBFRs is reviewed. Human exposure to NBFRs via different exposure pathways is discussed, and research gaps related to analysis, environmental sources, fate, and behaviour and human exposure are identified.


Expert Consensus to Guide the Classification of Paralympic Swimmers With Vision Impairment: A Delphi Study.
The International Paralympic Committee requires their member sports to develop a classification system that is sport-specific, meaning that the specific 'class' in which an athlete competes should be suitable for the degree to which the athlete's impairment affects performance in that particular sport. However, swimmers with vision impairment (VI) currently compete in classes that were developed on the basis of legal definitions of blindness, failing to consider how vision impacts swimming performance. The aim of this study was to establish expert guidance on the specific requirements for a sport-specific system of classification for VI swimming. A three-round Delphi review was conducted with a panel of 16 people with expertise in VI swimming either as an athlete, coach, administrator, or scientist. There was clear consensus (86%) among the panel that the current classification system used for VI swimming fails to fulfill the aim of minimizing the impact of VI on the outcome of competition. Particularly, the panel agreed that there are a range of aspects of visual function (e.g., depth perception and contrast sensitivity) that are important for optimal swimming performance, yet are not assessed using the current classification system. The panel also identified nine performance components of a swimming race that are mostly likely to be affected by VI. Interestingly, these were spread across all four major segments (start, clean swim, turn, and finish), and weren't necessarily those performance determinants generally used by performance analysts and coaches. There was also strong agreement that the age at which VI is acquired will substantially impact the ability of a swimmer to reach their full potential in the pool. The main implication is that changes are required to the way that swimmers with VI are classified for para-sport competition. Clear guidance has been provided for how to further the development of an evidence-based classification system.


Enolase is present in the cell wall of Saccharomyces cerevisiae.
Non-covalently attached or soluble cell wall proteins of Saccharomyces cerevisiae were extracted using a high pH/2-mercaptoethanol procedure and were separated for peptide sequencing using 2D-PAGE. A partial N-terminal sequence of a major protein spot was obtained and showed high identity with enolase gene products. Western blotting with an anti-enolase antibody confirmed that enolase was present in the cell wall extract. Biotinylation of cells prior to protein extraction with a membrane impermeable biotinylating agent confirmed that the detection was not owing to cell lysis during extraction. Transmission immunoelectron microscopy showed enolase to be present in the cell wall. Enolase contains no known secretion signal that would localize it to the cell wall. Thus S. cerevisiae must have further mechanisms for targeting proteins to the cell wall.


The in vitro effect of aqueous extract of Nigella sativa seeds on nitric oxide production.
The in vitro effect of aqueous extract of Nigella sativa seeds on nitric oxide (NO) production by murine macrophages was studied. Murine peritoneal macrophages were pre-incubated with the extract and then activated with Escherichia coli lipopolysaccharride. NO production was measured after 24 hours by spectrophotometry. The plant extract caused a dose-dependent decrease in NO production. Dialyzed preparation of the extract did not affect NO production. However, the boiled fraction of the extract resulted in a dose-dependent inhibition of NO apparently comparable to that of the whole extract. These results indicate that the aqueous extract of N. sativa seeds exhibits an inhibitory effect on nitric oxide production by murine macrophages and the active component(s) is/are non-protein in nature. In view of the fact that nitric oxide is a pro-inflammatory mediator, this study validates the traditional use of the Nigella sativa seeds for the treatment of rheumatism.


Regulation of the replication of influenza virus RNA segments: partial suppression of protein synthesis restores the 'early' replication pattern.
The replication of influenza virus RNA was monitored by RNA-RNA hybridization with subsequent analysis of hybrid duplexes, as well as by immunosorption of viral nucleocapsids from extracts of [3H]uridine-labelled cells followed by the isolation and characterization of nucleocapsid-associated RNA. The nucleocapsid-associated RNA preparations contained mostly negative-strand genomic RNA. Electrophoresis of the hybrid RNA duplexes or single-stranded nucleocapsid-associated RNA in polyacrylamide gel revealed an 'early' replication pattern, with a predominance of the NP and NS gene segments, in cells labelled from 0 to 1 h post-infection. At later stages of infection the pattern changed to the 'late' one, with the M gene segment in excess of NS, and the NP gene no longer predominant. Cycloheximide added as late as 2 or 3 h post-infection suppressed RNA replication. Moderate concentrations of cycloheximide inhibited the replication of NS and NP gene segments to a lesser degree than the replication of the other RNA segments, thus restoring the 'early' replication pattern. Cycloheximide treatment resulted in a slight increase in the percentage of positive strands in nucleocapsid-associated RNA. The role of protein synthesis in the transition from the 'early' to the 'late' pattern of influenza virus RNA synthesis is discussed.


Enhanced B7-2 gene expression by interferon-gamma in human monocytic cells is controlled through transcriptional and posttranscriptional mechanisms.
B7-2 is a costimulatory molecule expressed on professional antigen-presenting cells that provides T cells with a critical signal resulting in T-cell activation. Interferon-gamma (IFN-gamma) enhances B7-2 protein expression in monocytic cells. However, the molecular mechanisms controlling the enhanced expression of B7-2 are poorly understood. Northern blot and flow cytometry analysis revealed that human monocytes and the human monocytic cell line MonoMac6 (MM6) constitutively expressed B7-2 mRNA and protein and IFN-gamma treatment further enhanced the expression of both molecules. The ability of IFN-gamma to enhance B7-2 mRNA was evident at the dose of 31 U/mL and reached plateau levels at 500 U/mL. The effects of IFN-gamma on B7-2 mRNA expression were time dependent and occurred within 3 hours of treatment and increased through 24 hours. In vitro transcription assays and mRNA stability experiments showed that IFN-gamma increases both transcriptional activity and the stability of B7-2 mRNA. Treatment of MM6 cells with cycloheximide showed that de novo protein synthesis was not required for the IFN-gamma-enhanced expression of B7-2 mRNA. Overall, these studies show for the first time that IFN-gamma-enhanced expression of B7-2 protein in human monocytic cells is controlled at the gene level through a dual mechanism involving transcriptional and posttranscriptional mechanisms.


The intentional mentor: effective mentorship of undergraduate science students.
Promoting quality mentorship of undergraduate science students has recently emerged as an important strategy for successfully recruiting and retaining students in the sciences. Although numerous faculty members are naturally gifted mentors, most faculty are inserted into a mentorship role with little, if any, training. Successfully mentoring undergraduate science students requires a myriad of skills that can be honed with forethought and practice. In this essay, the value of mentoring, the developmental profile of young adult students, and the traits of a good mentor are explored. The Triangular Model proposed by W. Brad Johnson provides a theoretical framework for the development of effective mentorship. Fifteen tips gleaned from the literature and the author's personal experience are provided to help improve mentoring skills of faculty working with undergraduate science students.


Receptor-ligand requirements for increased NK cell polyfunctional potential in slow progressors infected with HIV-1 coexpressing KIR3DL1*h/*y and HLA-B*57.
Carriage of the natural killer (NK) receptor genotype KIR3DL1*h/*y with its HLA-B*57 ligand (*h/*y+B*57) is associated with slow time to AIDS and low viral load (VL). To provide a functional basis for these epidemiological observations, we assessed whether HIV-1-infected slow progressors (SP) carrying the *h/*y+B*57 compound genotype would have increased NK cell polyfunctional potential in comparison to SP with other killer immunoglobulin-like receptor (KIR)/HLA compound genotypes and whether this enhanced polyfunctionality was dependent upon the coexpression of both KIR3DL1*h/*y and HLA-B*57. The functional potential of NK cells was investigated by stimulating peripheral blood mononuclear cells with HLA-devoid targets or single HLA transfectants. Multiparametric flow cytometry was used to detect NK cells with seven functional profiles representing all permutations of CD107a expression and gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) secretion. NK cells from individuals carrying KIR3DL1 receptor-HLA-Bw4 ligand pairs had greater trifunctional responses than those from KIR3DL1 homozygotes (hmz), who were Bw6 homozygotes. NK cells from subjects carrying the *h/*y+B*57 genotypes exhibited the highest trifunctional potential, and this was dependent on cocarriage of the NK receptor and its ligand. Trifunctional cells secreted more of each function tested on a per-cell basis than each corresponding monofunctional NK subset. Although VL influenced NK functionality, individuals with defined KIR/HLA genotypes exhibited differences in NK cell polyfunctionality that could not be accounted for by VL alone. The protective effect of HLA-B*57 on slow progression to AIDS and low VL may be mediated through its interaction with KIR3DL1 alleles to educate NK cells for potent activity upon stimulation.


Safety and efficacy of a hydrogel inlay with laser in situ keratomileusis to improve vision in myopic presbyopic patients: one-year results.
To study the safety and efficacy of implanting a hydrogel corneal inlay (Raindrop Near Vision Inlay) concurrently with performing laser in situ keratomileusis (LASIK) to treat myopic presbyopia and to compare the results with results of the same treatment in emmetropic and hyperopic patients. Two private clinics, Tijuana and Monterrey, Mexico. Prospective nonrandomized clinical trial. Bilateral myopic LASIK was performed and a corneal inlay was concurrently implanted in the nondominant eye under a flap created using a femtosecond laser. Primary safety outcomes were the retention of corrected distance (CDVA) and near (CNVA) visual acuities. Efficacy was evaluated on the basis of uncorrected near (UNVA), intermediate (UIVA), and distance (UDVA) visual acuities. A patient questionnaire was used to assess the preoperative and postoperative incidence of visual symptoms, the ability to perform common tasks with no correction, and patient satisfaction with vision. Thirty eyes were enrolled. At each postoperative visit, the mean CDVA and CNVA were within one half line of preoperative measurements and no eye lost 2 or more lines of CDVA. The mean binocular UDVA, UIVA, and UNVA were better than 20/25 Snellen at all postoperative visits. By 6 months, 93% of patients had a binocular Snellen acuity of 20/25 or better across all visual ranges. According to patient questionnaires, 1 year after surgery, visual symptoms were at preoperative levels, 98% of all visual tasks could be easily performed without correction, and 90% of patients were satisfied or very satisfied with their overall vision. A hydrogel corneal inlay with concurrent LASIK was safe and effective for treating myopic presbyopia. Drs. Garza and Chayet are consultants to and investigators for Revision Optics, Inc.


Cadmium accumulation in gill, liver, kidney and muscle tissues of common carp, Cyprinus carpio, and Nile tilapia, Oreochromis niloticus.
Accumulation of cadmium in gill, liver, muscle and kidney tissues of Cyprinus carpio and Oreochromis niloticus were investigated in fish exposed to 0.5 ppm cadmium over 1, 15 and 30 days under controlled laboratory conditions. Tissue accumulation of the metal was measured using Atomic Absorption Spectrophotometric techniques. Cadmium accumulation in gill, liver, kidney and muscle, tissues of C. carpio and O. niloticus exposed to metal for 1, 15 and 30 days increased significantly compared with the control group (p < 0.05), except muscle tissue of O. niloticus. A general increase was observed in Cd accumulation with increasing exposure periods. Highest metal accumulation was observed in kidney followed by liver, gill and muscle tissues in both species. Liver accumulation of Cd was higher in C. carpio than O. niloticus, whereas kidney accumulation of the metal was higher in O. niloticus than C. carpio.


The innervation of dorsoventrally reversed chick wings: evidence that motor axons do not actively seek out their appropriate targets.
In normal chick embryos the extensor (dorsal) muscles are innervated by motoneurones lying laterally in the motor horn, while flexor muscles are supplied by more medially placed motoneurones. After reversal of the dorsoventral axis of the forelimb prior to innervation in most cases the opposite pattern is found, the extensors innervated by medial and flexors by lateral motor neurones. In a minority of cases the normal innervation pattern is obtained. Three hypotheses are discussed, two involving specific target affinity between motor axon and target and one involving passive deployment of axons to targets. We conclude that our results favour the latter hypothesis but that we cannot exclude a short-range specific signal.


Human DNA methyltransferase 1 is required for maintenance of the histone H3 modification pattern.
DNA methyltransferase 1 (DNMT1) plays an essential role in murine development and is thought to be the enzyme primarily responsible for maintenance of the global methylation status of genomic DNA. However, loss of DNMT1 in human cancer cells affects only the methylation status of a limited number of pericentromeric sequences. Here we show that human cancer cells lacking DNMT1 display at least two important differences with respect to wild type cells: a profound disorganization of nuclear architecture, and an altered pattern of histone H3 modification that results in an increase in the acetylation and a decrease in the dimethylation and trimethylation of lysine 9. Additionally, this phenotype is associated with a loss of interaction of histone deacetylases (HDACs) and HP1 (heterochromatin protein 1) with histone H3 and pericentromeric repetitive sequences (satellite 2). Our data indicate that DNMT1 activity, via maintenance of the appropriate histone H3 modifications, contributes to the preservation of the correct organization of large heterochromatic regions.


Prevalence and Treatment of Vitamin D Deficiency in Young Male Russian Soccer Players in Winter.
Vitamin D (25(OH)D) insufficiency and deficiency are highly prevalent in adult soccer players and can exceed 80% even in regions with high insolation; however, the treatment of this condition is often complicated. The aim of the present study was to examine the prevalence of vitamin D insufficiency and deficiency in youth Russian soccer players and the efficacy of its treatment. Participants were 131 young male football players (age 15.6 ± 2.4 years). Low vitamin D levels (below 30 ng/mL) were observed in 42.8% of the analyzed participants. These athletes were split in two groups composed of persons with vitamin D deficiency (serum vitamin D below 21 ng/mL) and insufficiency (serum vitamin D in range of 21-29 ng/mL). A dietary supplement of 5000 IU cholecalciferol per day was administered for two months. After the treatment, an average 92% increase in vitamin D concentration was observed (before treatment-19.7 ± 5.4 ng/mL, after treatment-34.7 ± 8.6 ng/mL, p<0.001) and 74% of the post-treatment values were within the reference range (30-60 ng/mL). Serum concentration of vitamin D increased by 200% ± 98% (p<0.001) during the first month of treatment with vitamin D deficiency and insufficiency being successfully treated in 83% of the football players. In summary, the prevalence of vitamin D insufficiency and deficiency was high in young Russian soccer players. Furthermore, it was indicated that the daily usage of cholecalciferol in a dose 5000 IU was an effective and well-tolerated treatment for vitamin D insufficiency. No linear dependency between the duration of treatment and increase in vitamin 25(OH)D concentration was observed.


Bilateral internal thoracic artery harvest and deep sternal wound infection in diabetic patients.
Coronary artery bypass graft surgery is superior to percutaneous intervention in diabetic patients with multivessel disease. The use of bilateral internal thoracic arteries (BITA) may provide better long-term graft patency, but the risk of postoperative deep sternal wound infection has limited its use in diabetic patients. However, studies have reported conflicting results, and require systematic evaluation. MEDLINE, EMBASE, World of Science, and the Cochrane library were searched for randomized controlled trials and observational studies comparing the incidence of deep sternal wound infection in diabetic patients undergoing either left internal thoracic artery (LITA) or BITA harvest. We used random effect models to compare risk ratios within groups. One randomized controlled trial and 10 observational studies (126,235 diabetic patients: 122,465 LITA, 3,770 BITA) met inclusion criteria. Deep sternal wound infection occurred in 3.1% and 1.6% for the BITA and LITA cohorts, respectively. The risk ratio for deep sternal wound infection development was 1.71 (1.37 to 2.14) for BITA compared with LITA. Patients who underwent skeletonized BITA harvest had a similar risk of deep sternal wound infection compared with LITA (0.9 [0.42 to 2.09]), although pedicled harvest demonstrated increased risk (1.77 [1.4 to 2.23]). Early mortality was comparable in the LITA cohort (2.5%) and the BITA cohort (2.3%; p = 0.8). The risk of deep sternal wound infection can be minimized in diabetic patients undergoing coronary artery bypass graft surgery by performing ITA harvested in a skeletonized manner with meticulous attention to preserving sternal blood flow. Pedicled harvest is to be discouraged when utilizing both ITA owing to a significant increase in the risk of postoperative deep sternal wound infection.


3D-Printed Antibacterial Scaffolds for the Regeneration of Alveolar Bone in Severe Periodontitis.
Current clinical treatment of periodontitis alleviates periodontal symptoms and helps to keep the disease under control for extended periods. Despite this, a significant destruction of the tooth's underlying bone tissue often takes place progressively. Herein, we present a two-way therapeutic approach for local delivery of antibacterial agents and bone tissue regeneration, incorporating ~1% w/w tetracycline hydrochloride (TCH) into a 3D-printed scaffold composed of poly(ε-caprolactone) (PCL). Samples were assessed for their morphological, physicochemical, pharmacokinetic, and antibacterial properties. Furthermore, osteoprecursor cells (MC3T3-E1) were employed to evaluate the osteoinductive potential of the drug-loaded scaffolds. Cell proliferation, viability, and differentiation were determined on all cell-seeded scaffolds. At the end of the culture, PCL-TCH scaffolds promoted abundant collagen organic matrix, demonstrating augmented alkaline phosphatase (ALP) activity and areas of accumulated mineralised bone tissue, despite their belayed cell proliferation. Based on the observed effectiveness of the PCL-TCH scaffolds to inhibit Staphylococcus aureus, these constructs could serve as an alternative bioactive implant that supports bacterial inhibition and favours a 3D microenvironment for bone tissue regeneration in severe periodontitis.


A mutant HBs antigen (HBsAg)183-191 epitope elicits specific cytotoxic T lymphocytes in acute hepatitis B patients.
HBs antigen (HBsAg)183-191 (FLLTRILTI, R187 peptide) is a dominant human leucocyte antigen-A2 (HLA-A2)-restricted epitope associated with hepatitis B virus (HBV) infection in Caucasian populations. However, its prevalence is poorly understood in China, where there is a high incidence of HBV infection. In this report, we sequenced the region of HBsAg derived from 103 Chinese patients. Approximately 16.5% of the patients bore a mutant HBsAg183-191 epitope in which the original arginine (R187) was substituted with a lysine (K187 mutant peptide). Importantly, K187 still bound to HLA-A2 with high affinity, and elicited specific cytotoxic T lymphocyte (CTL) responses in HLA-A2/K(b) transgenic mice. K187-specific CTLs were also generated successfully in acute hepatitis B (AHB) patients, indicating that this mutant epitope is processed and presented effectively. Our findings show that R187-specific CTLs can cross-react with the K187 peptide. These findings reveal that K187 still has the property of an HLA-A2 restricted epitope, and elicits a protective anti-HBV CTL response in humans.


Amebic meningoencephalitides and keratitis: challenges in diagnosis and treatment.
Acanthamoeba spp., Balamuthia mandrillaris, and Naegleria fowleri, although free-living amebae, also cause devastating diseases in humans leading to death. Acanthamoeba spp. and B. mandrillaris cause granulomatous amebic encephalitis, cutaneous and nasopharyngeal as well as disseminated infection. Acanthamoeba also causes a vision-threatening infection of the cornea, Acanthamoeba keratitis, principally in contact lens wearers. N. fowleri causes an acute, fulminating infection of the central nervous system, primary amebic meningoencephalitis, in healthy children and young adults who indulge in aquatic activities in fresh water. This review focuses on the recent developments in the diagnosis and treatment and clinical management of the diseases caused by these amebae. Development of a multiplex real-time PCR test has made it possible to simultaneously detect all the three free-living amebae in a sample. It is a rapid assay with a short turn-around time of just 4-5 h. An early diagnosis would be helpful in initiating potentially effective treatment. A recent study reported exciting results indicating that loading of rokitamycin in chitosan microspheres improves and prolongs the in-vitro anti-Acanthamoeba activity of the drug. Diagnoses of these infections are challenging and antimicrobial therapy is empirical, which often results in fatalities. Further research is needed to explore the possibility of a better drug delivery system that crosses the blood-brain barrier and effectively reach the central nervous system.


Higher-order reasoning training years after traumatic brain injury in adults.
To conduct a feasibility study to compare the effects of top-down Strategic Memory and Reasoning Training (SMART) versus information-based Brain Health Workshop (BHW, control) on gist-reasoning (ie, abstracting novel meaning from complex information), memory, executive functions, and daily function in adults with traumatic brain injury. Twenty-eight participants (of the 35 recruited), 16 men & 12 women, aged 20 to 65 years (M = 43, SD = 11.34) at chronic stages posttraumatic brain injury (2 years or longer) completed the training. Fourteen participants that received SMART and 14 participants that completed BHW were assessed both pre- and posttraining. Thirteen of the SMART trained and 11 from BHW participated in a 6-month testing. The study was a single blinded randomized control trial. Participants in both groups received a minimum of 15 hours of training over 8 weeks. The SMART group significantly improved gist-reasoning as compared to the BHW group. Benefits of the SMART extended to untrained measures of working memory and participation in functional activities. Exploratory analyses suggested potential transfer effects of SMART on memory and executive functions. The benefits of the SMART program as compared to BHW were evident at immediately posttraining and 6 months posttraining. This study provides preliminary evidence that short-term intensive training in top-down modulation of information benefits gist-reasoning and generalizes to measures of executive function and real life function at chronic stages of post-TBI.


Pathological and microbiological findings from incidents of unusual mortality of the common frog (Rana temporaria).
In 1992 we began an investigation into incidents of unusual and mass mortalities of the common frog (Rana temporaria) in Britain which were being reported unsolicited to us in increasing numbers by members of the public. Investigations conducted at ten sites of unusual mortality resulted in two main disease syndromes being found: one characterized by skin ulceration and one characterized by systemic haemorrhages. However, frogs also were found with lesions common to both of these syndromes and microscopic skin lesions common to both syndromes were seen. The bacterium Aeromonas hydrophila, which has been described previously as causing similar lesions, was isolated significantly more frequently from haemorrhagic frogs than from those with skin ulceration only. However, as many of the latter were euthanased, this may have been due to differences in post mortem bacterial invasion. An iridovirus-like particle has been identified on electron microscopical examination of skin lesions from frogs with each syndrome and iridovirus-like inclusions have been detected in the livers of frogs with systemic haemorrhages. Also, an adenovirus-like particle has been cultured from one haemorrhagic frog. A poxvirus-like particle described previously from diseased frogs has now been found also in control animals and has been identified as a melanosome. Both the prevalence of the iridovirus-like particle and its association with lesions indicate that it may be implicated in the aetiology of the disease syndromes observed. Specifically, we hypothesize that primary iridovirus infection, with or without secondary infection with opportunistic pathogens such as A. hydrophila, may cause natural outbreaks of 'red-leg', a disease considered previously to be due to bacterial infection only.


Synthesis and characterization of tetracycline-imprinted membranes: A detailed positron annihilation lifetime spectroscopy investigation.
Molecularly imprinted thin membranes for selective removal of tetracycline from real water samples were prepared using the radiation-induced polymerization method. The chemical and physical characterization of the membranes was conducted by FTIR, XPS, AFM and SEM analyses. The effect of the template on the size and the size distribution of the pores in the membrane structure was investigated by positron annihilation lifetime spectroscopy (PALS) experiments. The presence of the template molecule causes the formation of larger cavities, which have a strong correlation to the molecular size of the template molecule. The density of the free volume holes in the imprinted network shows a significant increase due to the presence of the template molecule. The binding performances of the membranes were tested against various factors such as pH, time and initial concentration of template molecules. The specific selectivity of the membranes was investigated by cross-reactivity experiments. The binding capacity of the imprinted membranes was obtained at 14.5 μmol g-1 , with the highest imprinting factor (2.84) for tetracycline. The imprinting factor was obtained at 1.44, 1.25 and 1.57 for oxytetracycline hydrochloride, doxycycline hyclate and chlortetracycline, respectively. The binding capability of the imprinted membranes in real water samples was promising for the application of the membranes as a filter material for removal of tetracycline with high binding capacity. Tetracycline binding percentage of the membranes was determined at 92.4, 81.1., 75.0 and 68.7 for ultra-pure water, tap water and natural water samples collected from different sources, respectively.


Detection of DNA and RNA virus genomes in organ systems of whole mice: patterns of mouse organ infection by polyomavirus.
A technique which detects viral DNA or RNA in situ in the organ systems of whole mice is described. Frozen thin sections from whole mice were transferred directly to nitrocellulose and hybridized to labeled viral DNA, allowing the detection of viral DNA or RNA. By this procedure, polyomavirus infection of newborn mice inoculated intranasally was followed. We found that the initial inoculum could be detected in the nasal cavity, lungs, and stomach lining after a 5-h absorption period. Primary replication of virus was observed in the nasal cavity, submaxillary gland, and lungs, followed by a systemic phase of infection in which the liver, spleen, kidney, and large colon also became infected. Viral RNA as well as DNA could also be detected as shown by infecting mice intracerebrally with vesicular stomatitis virus. Vesicular stomatitis virus-specific RNA was observed only in the brains of these mice. It is most likely that this technique can be applied to general molecular studies of mice. With this method we should be able to detect all viruses, bacteria, plasmids, and organ-specific transcripts to which a cloned probe exists.