Tobacco and alcohol-related interventions for people with mild/moderate intellectual disabilities: a systematic review of the literature.
The behavioural determinants of health among people with mild/moderate intellectual disabilities (ID) are of increasing concern. With the closure of long-stay institutions, more people with ID are living in the community. As they lead more ordinary and less restricted lives, people with ID may be exposed to social and environmental pressures that encourage them to adopt behaviours that impact negatively on their health. Levels of smoking and alcohol consumption in this client group are of particular concern. We undertook a mixed method review of the literature, aiming to assess the Feasibility, Appropriateness, Meaningfulness and Effectiveness (FAME) of interventions designed to address the use of tobacco and/or alcohol in people with mild/moderate ID. Key electronic databases were searched (e.g., Medline, Cochrane Register of Controlled Trials, PsycINFO) from 1996 to 2011. The search was developed using appropriate subject headings and key words (e.g., intellectual disability, tobacco use, alcohol drinking, health promotion). On completion of the database searches, inclusion/exclusion criteria, based on an adaptation of the PICO framework (Population, Intervention, Comparison, Outcomes), were applied. Methodological quality was assessed using a seven-point rating scale. Database searches identified 501 unique records, of which nine satisfied the inclusion criteria. Four focused on tobacco, three on alcohol and two on both tobacco and alcohol. Located in the U.K., the U.S.A. and Australia, the studies aimed to increase knowledge levels and/or change behaviour (e.g., to encourage smoking cessation). One was a randomised controlled trial, one a quasi-experiment and the others were before and after studies and/or case studies. Methodological quality was poor or moderate. The combined studies had a sample size of 341, with ages ranging from 14 to 54 years. The interventions were delivered by professionals (e.g., in health, social care, education) during sessions that spanned a period of three weeks to one academic year. The studies highlighted a number of important issues linked to the appropriateness of interventions for this client group (e.g., use of pictures, quizzes, role play, incentives); however, in the majority of cases the interventions appeared to lack a theoretical framework (e.g., behaviour change theory). The appropriateness of the outcome measures for use with this client group was not tested. One study discussed feasibility (teachers delivering lessons on alcohol and tobacco) and only one was informative in terms of effectiveness, i.e., increasing knowledge of the health and social dangers of smoking and excessive alcohol consumption. This review is the first to systematically collate evidence on tobacco and alcohol-related interventions for people with ID. While there is currently little evidence to guide practice, the review delivers clear insights for the development of interventions and presents a strong case for more robust research methods. In particular there is a need to test the effectiveness of interventions in large-scale, well-designed trials and to ensure that outcome measures are developed/tailored appropriately for this client group.


Improving spine surgical access, appropriateness and efficiency in metropolitan, urban and rural settings.
The Inter-professional Spine Assessment and Education Clinics (ISAEC) were developed to improve primary care assessment, education and management of patients with persistent or recurrent low back pain-related symptoms. This study aims to determine the effect of ISAEC on access for surgical assessment, referral appropriateness and efficiency for patients meeting a priori referral criteria in rural, urban and metropolitan settings. We conducted a retrospective review of prospective data from networked ISAEC clinics in Thunder Bay, Hamilton and Toronto, Ontario. For patients meeting surgical referral criteria, wait times for surgical assessment, surgical referral-related magnetic resonance imaging (MRI) scans and appropriateness of referral were recorded. Overall 422 patients, representing 10% of all ISAEC patients in the study period, were referred for surgical assessment. The average wait times for surgical assessment were 5.4, 4.3 and 2.2 weeks at the metropolitan, urban and rural centres, respectively. Referral MRI usage for the group decreased by 31%. Of the patients referred for formal surgical assessment, 80% had leg-dominant pain and 96% were deemed appropriate surgical referrals. Contrary to geographic concentration of health care resources in metropolitan settings, the greatest decrease in wait times was achieved in the rural setting. A networked, shared-cared model of care for patients with low back pain-related symptoms significantly improved access for surgical assessment despite varying geographic practice settings and barriers. The greatest reductions were noted in the rural setting. In addition, significant improvements in referral appropriateness and efficiency were achieved compared with historical reports across all sites.


Repeat polymorphisms in the Homo sapiens heme oxygenase-1 gene in diabetic and idiopathic gastroparesis.
Idiopathic and diabetic gastroparesis in Homo sapiens cause significant morbidity. Etiology or risk factors have not been clearly identified. Failure to sustain elevated heme oxygenase-1 (HO1) expression is associated with delayed gastric emptying in diabetic mice and polymorphisms in the HO1 gene (HMOX1, NCBI Gene ID:3162) are associated with worse outcomes in other diseases. Our hypothesis was that longer polyGT alleles are more common in the HMOX1 genes of individuals with gastroparesis than in controls without upper gastrointestinal motility disorders. Repeat length was determined in genomic DNA. Controls with diabetes (84 type 1, 84 type 2) and without diabetes (n = 170) were compared to diabetic gastroparetics (99 type 1, 72 type 2) and idiopathic gastroparetics (n = 234). Correlations of repeat lengths with clinical symptom sub-scores on the gastroparesis cardinal symptom index (GCSI) were done. Statistical analyses of short (<29), medium and long (>32) repeat alleles and differences in allele length were used to test for associations with gastroparesis. The distribution of allele lengths was different between groups (P = 0.016). Allele lengths were longest in type 2 diabetics with gastroparesis (29.18±0.35, mean ± SEM) and longer in gastroparetics compared to non-diabetic controls (28.50±0.14 vs 27.64±0.20 GT repeats/allele, P = 0.0008). Type 2 diabetic controls had longer alleles than non-diabetic controls. In all gastroparetic groups, allele lengths were longer in African Americans compared to other racial groups, differences in the proportion of African Americans in the groups accounted for the differences between gastroparetics and controls. Diabetic gastroparetics with 1 or 2 long alleles had worse GCSI nausea sub-scores (3.30±0.23) as compared to those with 0 long alleles (2.66±0.12), P = 0.022. Longer poly-GT repeats in the HMOX1 gene are more common in African Americans with gastroparesis. Nausea symptoms are worse in subjects with longer alleles.


How receptors talk to trimeric G proteins.
Stimulated by hormones and sensory stimuli, serpentine receptors promote the release of GDP that is bound to the alpha subunit of trimeric G proteins and its replacement by GTP. Recent investigations have begun to define the sizes, shapes, and relative orientations of receptors and G proteins, the surfaces through which they interact with one another, and conformational changes in both sets of molecules that underlie receptor-catalyzed guanine-nucleotide exchange.


Biphasic voltage-dependent inactivation of human NaV 1.3, 1.6 and 1.7 Na+ channels expressed in rodent insulin-secreting cells.
Na+ current inactivation is biphasic in insulin-secreting cells, proceeding with two voltage dependences that are half-maximal at ∼-100 mV and -60 mV. Inactivation of voltage-gated Na+ (NaV ) channels occurs at ∼30 mV more negative voltages in insulin-secreting Ins1 and primary β-cells than in HEK, CHO or glucagon-secreting αTC1-6 cells. The difference in inactivation between Ins1 and non-β-cells persists in the inside-out patch configuration, discounting an involvement of a diffusible factor. In Ins1 cells and primary β-cells, but not in HEK cells, inactivation of a single NaV subtype is biphasic and follows two voltage dependences separated by 30-40 mV. We propose that NaV channels adopt different inactivation behaviours depending on the local membrane environment. Pancreatic β-cells are equipped with voltage-gated Na+ channels that undergo biphasic voltage-dependent steady-state inactivation. A small Na+ current component (10-15%) inactivates over physiological membrane potentials and contributes to action potential firing. However, the major Na+ channel component is completely inactivated at -90 to -80 mV and is therefore inactive in the β-cell. It has been proposed that the biphasic inactivation reflects the contribution of different NaV α-subunits. We tested this possibility by expression of TTX-resistant variants of the NaV subunits found in β-cells (NaV 1.3, NaV 1.6 and NaV 1.7) in insulin-secreting Ins1 cells and in non-β-cells (including HEK and CHO cells). We found that all NaV subunits inactivated at 20-30 mV more negative membrane potentials in Ins1 cells than in HEK or CHO cells. The more negative inactivation in Ins1 cells does not involve a diffusible intracellular factor because the difference between Ins1 and CHO persisted after excision of the membrane. NaV 1.7 inactivated at 15--20 mV more negative membrane potentials than NaV 1.3 and NaV 1.6 in Ins1 cells but this small difference is insufficient to solely explain the biphasic inactivation in Ins1 cells. In Ins1 cells, but never in the other cell types, widely different components of NaV inactivation (separated by 30 mV) were also observed following expression of a single type of NaV α-subunit. The more positive component exhibited a voltage dependence of inactivation similar to that found in HEK and CHO cells. We propose that biphasic NaV inactivation in insulin-secreting cells reflects insertion of channels in membrane domains that differ with regard to lipid and/or membrane protein composition.


Angiogenic potential of the cerebrospinal fluid (CSF) of patients with high-grade gliomas measured with the chick embryo chorioallantoic membrane assay (CAM).
High-grade gliomas are highly vascularized tumors. Neo-angiogenesis plays a key role in tumor growth and resistance to therapy. A cerebrospinal fluid (CSF) sample could be a useful way to obtain pro-angiogenic predictive or prognostic markers at different stages of the disease. As a first step we looked for pro-angiogenic activity in the CSF of patients with high-grade gliomas. We performed the chicken embryo chorio-allantoic membrane (CAM) assay to study the angiogenic potential of the cerebrospinal fluid (CSF), obtained either by lumbar puncture (LP) or craniotomy from six patients with high-grade brain tumors (three glioblastoma (WHO grade IV), one anaplastic oligodendroglioma (WHO grade III), two anaplastic ganglioglioma (WHO grade III)), and four healthy controls. Significantly increased neo-angiogenesis was observed on the surface of the growing CAM in the 6 patients with high-grade gliomas compared to controls (3.69 ± 1.23 versus 2.16 ± 0.97 capillaries per area (mean ± SD), p<0.005). There was no statistical difference related to the hystological grade of the tumor (WHO grade III or IV), previous treatment (radio-chemotherapy plus temozolomide, temozolomide alone or no treatment), or the site of CSF sample (surgery or lumbar puncture). Our results suggest a pro-angiogenic potential in the CSF of patients with high-grade gliomas.


Ecosystem productivity is associated with bacterial phylogenetic distance in surface marine waters.
Understanding the link between community diversity and ecosystem function is a fundamental aspect of ecology. Systematic losses in biodiversity are widely acknowledged but the impact this may exert on ecosystem functioning remains ambiguous. There is growing evidence of a positive relationship between species richness and ecosystem productivity for terrestrial macro-organisms, but similar links for marine micro-organisms, which help drive global climate, are unclear. Community manipulation experiments show both positive and negative relationships for microbes. These previous studies rely, however, on artificial communities and any links between the full diversity of active bacterial communities in the environment, their phylogenetic relatedness and ecosystem function remain hitherto unexplored. Here, we test the hypothesis that productivity is associated with diversity in the metabolically active fraction of microbial communities. We show in natural assemblages of active bacteria that communities containing more distantly related members were associated with higher bacterial production. The positive phylogenetic diversity-productivity relationship was independent of community diversity calculated as the Shannon index. From our long-term (7-year) survey of surface marine bacterial communities, we also found that similarly, productive communities had greater phylogenetic similarity to each other, further suggesting that the traits of active bacteria are an important predictor of ecosystem productivity. Our findings demonstrate that the evolutionary history of the active fraction of a microbial community is critical for understanding their role in ecosystem functioning.


Highly effective hydrogen isotope separation in nanoporous metal-organic frameworks with open metal sites: direct measurement and theoretical analysis.
Separating gaseous mixtures that consist of very similar size is one of the critical issues in modern separation technology. Especially, the separation of the isotopes hydrogen and deuterium requires special efforts, even though these isotopes show a very large mass ratio. Conventionally, H/D separation can be realized through cryogenic distillation of the molecular species or the Girdler-sulfide process, which are among the most energy-intensive separation techniques in the chemical industry. However, costs can be significantly reduced by using highly mass-selective nanoporous sorbents. Here, we describe a hydrogen isotope separation strategy exploiting the strongly attractive open metal sites present in nanoporous metal-organic frameworks of the CPO-27 family (also referred to as MOF-74). A theoretical analysis predicts an outstanding hydrogen isotopologue separation at open metal sites due to isotopal effects, which has been directly observed through cryogenic thermal desorption spectroscopy. For H2/D2 separation of an equimolar mixture at 60 K, the selectivity of 12 is the highest value ever measured, and this methodology shows extremely high separation efficiencies even above 77 K. Our theoretical results imply also a high selectivity for HD/H2 separation at similar temperatures, and together with catalytically active sites, we propose a mechanism to produce D2 from HD/H2 mixtures with natural or enriched deuterium content.


Antihypertensive effect of Pleurotus nebrodensis in spontaneously hypertensive rats.
In this study, we examined the effects of Pleurotus nebrodensis on systolic blood pressure of spontaneously hypertensive rats. Single-dose and continuous-dose tests with sample diets made from the fruit body of the mushroom, P. nebrodensis were carried out on SHR and control rats. Sample diets included 6% dry powder of fruit body (6% dry powder), hot water extract, polysaccharide fraction, protein fraction, dialyzable fraction and non-dialyzable fraction. Polysaccharide and protein fractions were obtained by hot water extraction. The protein fraction was separated to the dialyzable fraction and non-dialyzable fraction by dialysis. In the single-dose test, protein fraction, hot water extract and polysaccharide fraction decreased systolic blood pressure. Blood pressure was lowered after administration for 2 h, and it returned to the pre-administration blood pressure after 48 h. In the continuous-dose test, spontaneously hypertensive rats were fed each of the diets for 16 weeks. The 6% dry powder group showed significantly inhibited elevation of blood pressure compared with the control group and there was no influence on total cholesterol and triglyceride levels. The non-dialyzable fraction showed suppression of increase in blood pressure from the start of the continuous oral administration. Effects on the rennin angiotensin system and renal function were also indicated. The antihypertensive action effect of P. nebrodensis can be expected to not only prevent but also to improve hypertension.


Stimulation of DNA polymerase alpha activity by microtubule-associated proteins.
Microtubule-associated protein 2 (MAP2) isolated from porcine brains stimulated the activity of DNA polymerase alpha immunopurified from calf thymus or human lymphoma cells, in a dose-dependent manner. This stimulation was pronounced when activated DNA or poly(dA).(dT)10 was used as the template-primer. DNA polymerase alpha bound to a MAP2-immobilized column, whereas preincubation of the enzyme with unbound MAP2 prevented binding to the column. These events suggested that a physical binding occurred between the polymerase and MAP2. Kinetic analyses revealed that MAP2 decreased the Km value of the polymerase for deoxyribonucleotides, irrespective of the species of template-primer. A concomitant increase in Vmax was observed; however, the extent of the increase depended on the species of template-primer. MAP2 also decreased the Km value of the polymerase for template-primers when activated DNA of poly(dA).(dT)10 was used as the template-primer. Product analyses showed that MAP2 did not significantly alter the processivity of the polymerase and the increment of Vmax is considered to be due to an increase in the frequency of initiation of DNA synthesis. The stimulation by MAP2 occurred specifically in the activity of DNA polymerase alpha, but not DNA polymerases beta, gamma, and I from Escherichia coli. Other MAPs, tau and 190-kDa MAP, could substitute for MAP2. Thus, the specific stimulation of DNA polymerase alpha by MAPs supports the notion of a possible involvement of MAPs or MAP-like proteins in DNA replication, in vivo.


Brain MR in Fukuyama congenital muscular dystrophy.
To determine the MR characteristics of brain abnormalities in Fukuyama congenital muscular dystrophy (FCMD). We reviewed 30 MR examinations of 21 patients with FCMD to assess cerebral and cerebellar cortical dysplasia, white matter changes, and miscellaneous abnormalities. On MR images, all patients had thick and bumpy cortices with shallow sulci corresponding to polymicrogyria, and 12 patients had pachygyric cortices with smooth surfaces, corresponding to type II lissencephaly. Both types of cortical dysplasia had characteristic distributions: the first type involved the frontal lobe in all 21 patients and also the parietotemporal lobe in 6 patients; the second type involved the temporooccipital lobes. Eighteen patients had prolonged T1 and T2 signal in the white matter, which was indistinct in neonates and seen infrequently in adolescents. In four patients, abnormal vessels were seen within the pachygyric cortices. MR studies of the brain show findings consistent with the known characteristics of FCMD. The MR detection of the two types of cerebral cortical dysplasia with characteristic distribution and cerebellar abnormalities is helpful in the differential and early diagnosis of FCMD.


Expression profiling of mammary carcinoma cell lines: correlation of in vitro invasiveness with expression of CD24.
Invasiveness and the capacity of tumor cells to form distant metastases are important cellular characteristics associated with a poor prognosis in breast cancer patients. In an approach to find genes that are potentially involved in these processes, RNA species showing different abundance in RNA pools from 12 invasive and 13 noninvasive mammary carcinoma-derived cell lines have been identified by hybridization to cDNA microarrays. CD24, keratin 19, keratin 8, GOB-4 and ezrin-radixin-moesin-binding phosphoprotein 50 were found to be preferentially expressed by noninvasive cells whereas vimentin was confirmed as a characteristic of invasive cells. Only differences in expression higher than 3-fold evident in three independent hybridization experiments were considered significant. For all cell lines, expression of mRNA coding for the adhesion molecule CD24, previously suggested to play an important role during tumor progression to more invasive phenotypes, has been quantified by real-time RT-PCR. Flow-cytometric analyses confirmed that CD24 mRNA reflects the amount of cell surface CD24 (Spearman R = 0.88, p = 10(-6)). CD24 mRNA was found to be absent or weakly expressed in 9/12 (75%) invasive cell lines compared to 3/13 (23%) noninvasive cell lines. The correlation between CD24 expression and invasiveness was calculated to be highly significant with chi2 = 6.74 and p = 0.0094. Future analyses of primary breast carcinomas are warranted to define the role of CD24 in future diagnostic and therapeutic approaches.


Computer simulations of protein functions: searching for the molecular origin of the replication fidelity of DNA polymerases.
The use of computers to simulate the functions of complex biological macromolecules is essential to achieve a microscopic description of biological processes and to model and interpret experimental data. Here we apply theoretical computational approaches to investigate the fidelity of T7 DNA polymerase, divided into discrete steps that include contributions from substrate binding, pK(a) shifts, and rate constants for the PO bond-breaking and bond-making processes. We begin by defining the discrimination between right and wrong nucleotides in terms of the free energy landscape for the dNMP incorporation reaction. We then use the linear response approximation and the empirical valence bond methods to obtain converging results for the contribution of the binding and chemical steps to the overall fidelity. These approaches are successful in reproducing general trends in the observed polymerase incorporation fidelity. The calculations demonstrate the potential for further integration of theoretical and experimental studies to analyze high- and low-fidelity DNA polymerases.


Nursing responses to transcultural encounters: what nurses draw on when faced with a patient from another culture.
We explored nurses' experiences when they encounter patients from cultures other than their own and their perception of what helps them deliver culturally competent care. Registered nurses from all shifts and units at Kaiser Permanente Santa Clara Medical Center were invited to complete a questionnaire. Within the time frame allowed, 111 nurses participated by returning completed questionnaires.A descriptive survey was conducted using a questionnaire that contained multiple-choice, fill-in-the-blank, and open-ended items. A large majority of respondents reported that they drew on prior experience, including experience with friends and family, and through their education and training, and more than half also included travel experience and information obtained through the Internet and news media. They also expressed a desire for more training and continuing education, exposure to more diverse cultures, and availability of more interpreters. When respondents were asked to enumerate the cultures from which their patients have come, their answers were very specific, revealing that these nurses understood culture as going beyond ethnicity to include religious groups, sexual orientation, and social class (eg, homeless). Our research confirmed our hypothesis that nurses are drawing heavily on prior experience, including family experiences and experiences with friends and coworkers from different cultures. Our findings also suggest that schools of nursing are providing valuable preparation for working with diverse populations. Our research was limited to one geographic area and by our purposeful exclusion of a demographic questionnaire. We recommend that this study be extended into other geographic areas. Our study also shows that nurses are drawing on their experiences in caring for patients from other cultures; therefore, we recommend that health care institutions consider exposing not only nurses but also other health care professionals to different cultures by creating activities that involve community projects in diverse communities, offering classes or seminars on different cultures and having an active cultural education program that would reach out to nurses. The experiences provided by such activities and programs would help nurses become more sensitive to the differences between cultures and not immediately judge patients or make assumptions about them.


Genetic analysis of self-associating immunoglobulin G rheumatoid factors from two rheumatoid synovia implicates an antigen-driven response.
Although much has been learned about the molecular basis of immunoglobulin M (IgM) rheumatoid factors (RFs) in healthy individuals and in patients with mixed cryoglobulinemia and rheumatoid arthritis, little is known about the genetic origins of the potentially pathogenic IgG RFs in the inflamed rheumatoid synovia of patients. Recently, we generated from unmanipulated synovium B cells several hybridomas that secreted self-associating IgG RFs. To delineate the genetic origins of such potentially pathogenic RFs, we adapted the anchored polymerase chain reaction to rapidly clone and characterize the expressed Ig V genes for the L1 and the D1 IgG RFs. Then, we identified the germline counterparts of the expressed L1 IgG RF V genes. The results showed that the L1 heavy chain was encoded by a Vh gene that is expressed preferentially during early ontogenic development, and that is probably located within 240 kb upstream of the Jh locus. The overlap between this RF Vh gene and the restricted fetal antibody repertoire is reminiscent of the natural antibody-associated Vh genes, and suggests that at least part of the "potential pathogenic" IgG RFs in rheumatoid synovium may derive from the "physiological" natural antibody repertoire in a normal immune system. Indeed, the corresponding germline Vh gene for L1 encodes the heavy chain of an IgM RF found in a 19-wk-old fetal spleen. Furthermore, the comparisons of the expressed RF V genes and their germline counterparts reveal that the L1 heavy and light chain variable regions had, respectively, 16 and 7 somatic mutations, which resulted in eight and four amino acid changes. Strikingly, all eight mutations in the complementarity determining regions of the V gene-encoded regions were replacement changes, while only 6 of 11 mutations in the framework regions caused amino acid changes. Combined with L1's high binding affinity toward the Fc fragment, these results suggest strongly that the L1 IgG RF must have been driven by the Fc antigen.


Toxicological characterization of a novel in vivo benzo[a]pyrene metabolite, 7-oxo-benz[d]anthracene-3,4-dicarboxylic acid anhydride.
Recently, we described a new in vivo pathway in the metabolism of benzo[a]pyrene (BP) that involves an opening of the aromatic ring system. One of the products of this pathway, isolated from rat urine, was the anhydride of 7-oxo-benz[d]anthracene-3,4-dicarboxylic acid (ABADA). We have now investigated the effect of ABADA on several cellular targets, known to be important in tumor formation. ABADA was as efficient as BP-7,8-diol-9,10-epoxide in inducing direct strand breaks but not alkali labile sites in DNA in HT-29 cells and exhibited weak mutagenic activity in Salmonella typhimurium strain TA 102. The cytotoxicity of ABADA to HCT 116 cells appeared to be due to apoptosis, as caspase-3 activity and poly-ADP-ribose polymerase (PARP) cleavage was observed. COX-2 promoter activity was induced by ABADA in HCT 116 cells. In conclusion, this novel metabolic pathway may also be contributing to the carcinogenicity of BP.


Transcriptomic analysis reveals evidence for a cryptic plastid in the colpodellid Voromonas pontica, a close relative of chromerids and apicomplexan parasites.
Colpodellids are free-living, predatory flagellates, but their close relationship to photosynthetic chromerids and plastid-bearing apicomplexan parasites suggests they were ancestrally photosynthetic. Colpodellids may therefore retain a cryptic plastid, or they may have lost their plastids entirely, like the apicomplexan Cryptosporidium. To find out, we generated transcriptomic data from Voromonas pontica ATCC 50640 and searched for homologs of genes encoding proteins known to function in the apicoplast, the non-photosynthetic plastid of apicomplexans. We found candidate genes from multiple plastid-associated pathways including iron-sulfur cluster assembly, isoprenoid biosynthesis, and tetrapyrrole biosynthesis, along with a plastid-type phosphate transporter gene. Four of these sequences include the 5' end of the coding region and are predicted to encode a signal peptide and a transit peptide-like region. This is highly suggestive of targeting to a cryptic plastid. We also performed a taxon-rich phylogenetic analysis of small subunit ribosomal RNA sequences from colpodellids and their relatives, which suggests that photosynthesis was lost more than once in colpodellids, and independently in V. pontica and apicomplexans. Colpodellids therefore represent a valuable source of comparative data for understanding the process of plastid reduction in humanity's most deadly parasite.


Neural correlates of behavior in the moth Manduca sexta in response to complex odors.
With Manduca sexta as a model system, we analyzed how natural odor mixtures that are most effective in eliciting flight and foraging behaviors are encoded in the primary olfactory center in the brain, the antennal lobe. We used gas chromatography coupled with multiunit neural-ensemble recording to identify key odorants from flowers of two important nectar resources, the desert plants Datura wrightii and Agave palmeri, that elicited responses from individual antennal-lobe neurons. Neural-ensemble responses to the A. palmeri floral scent, comprising >60 odorants, could be reproduced by stimulation with a mixture of six of its constituents that had behavioral effectiveness equivalent to that of the complete scent. Likewise, a mixture of three floral volatiles from D. wrightii elicited normal flight and feeding behaviors. By recording responses of neural ensembles to mixtures of varying behavioral effectiveness, we analyzed the coding of behaviorally "meaningful" odors. We considered four possible ensemble-coding mechanisms--mean firing rate, mean instantaneous firing rate, pattern of synchronous ensemble firing, and total net synchrony of firing--and found that mean firing rate and the pattern of ensemble synchrony were best correlated with behavior (R = 41% and 43%, respectively). Stepwise regression analysis showed that net synchrony and mean instantaneous firing rate contributed little to the variation in the behavioral results. We conclude that a combination of mean-rate coding and synchrony of firing of antennal-lobe neurons underlies generalization among related, behaviorally effective floral mixtures while maintaining sufficient contrast for discrimination of distinct scents.


Artificial intelligence in inflammatory bowel disease: implications for clinical practice and future directions.
Inflammatory bowel disease encompasses Crohn's disease and ulcerative colitis and is characterized by uncontrolled, relapsing, and remitting course of inflammation in the gastrointestinal tract. Artificial intelligence represents a new era within the field of gastroenterology, and the amount of research surrounding artificial intelligence in patients with inflammatory bowel disease is on the rise. As clinical trial outcomes and treatment targets evolve in inflammatory bowel disease, artificial intelligence may prove as a valuable tool for providing accurate, consistent, and reproducible evaluations of endoscopic appearance and histologic activity, thereby optimizing the diagnosis process and identifying disease severity. Furthermore, as the applications of artificial intelligence for inflammatory bowel disease continue to expand, they may present an ideal opportunity for improving disease management by predicting treatment response to biologic therapies and for refining the standard of care by setting the basis for future treatment personalization and cost reduction. The purpose of this review is to provide an overview of the unmet needs in the management of inflammatory bowel disease in clinical practice and how artificial intelligence tools can address these gaps to transform patient care.


fMRI congruous word repetition effects reflect memory variability in normal elderly.
Neural circuits mediating repetition effect for semantically congruous words on functional MRI were investigated in seventeen normal elderly (mean age=70). Participants determined if written words were semantically congruent (50% probability) with spoken statements. Subsequent cued-recall revealed robust explicit memory only for congruous items (83% versus 8% for incongruous). Event-related BOLD responses to New>Old congruous words were found in the left>right cingulate and fusiform gyri, left parahippocampal cortex, middle and inferior frontal gyri (IFG). A group with above-median subsequent recall had markedly more widespread BOLD responses than a Low-Recall subgroup, with larger responses in the left medial temporal lobe (LMTL), IFG, and bilateral cingulate gyri. The magnitude of LMTL activation (New-Old) correlated with subsequent cued-recall, while the spatial extent of LMTL activation (New>Old) correlated with recall and recognition. Both magnitude and spatial extent of left fusiform activation correlated with subsequent recall/recognition. A neural circuit of left-hemisphere brain regions, many identified as P600 generators by invasive electrophysiological studies, was activated by New>Old congruous words, likely mediating successful verbal encoding.


Natural killer cell dependent within-host competition arises during multiple MCMV infection: consequences for viral transmission and evolution.
It is becoming increasingly clear that many diseases are the result of infection from multiple genetically distinct strains of a pathogen. Such multi-strain infections have the capacity to alter both disease and pathogen dynamics. Infection with multiple strains of human cytomegalovirus (HCMV) is common and has been linked to enhanced disease. Suggestions that disease enhancement in multi-strain infected patients is due to complementation have been supported by trans-complementation studies in mice during co-infection of wild type and gene knockout strains of murine CMV (MCMV). Complementation between naturally circulating strains of CMV has, however, not been assessed. In addition, many models of multi-strain infection predict that co-infecting strains will compete with each other and that this competition may contribute to selective transmission of more virulent pathogen strains. To assess the outcome of multi-strain infection, C57BL/6 mice were infected with up to four naturally circulating strains of MCMV. In this study, profound within-host competition was observed between co-infecting strains of MCMV. This competition was MCMV strain specific and resulted in the complete exclusion of certain strains of MCMV from the salivary glands of multi-strain infected mice. Competition was dependent on Ly49H(+) natural killer (NK) cells as well as the expression of the ligand for Ly49H, the MCMV encoded product, m157. Strains of MCMV which expressed an m157 gene product capable of ligating Ly49H were outcompeted by strains of MCMV expressing variant m157 genes. Importantly, within-host competition prevented the shedding of the less virulent strains of MCMV, those recognized by Ly49H, into the saliva of multi-strain infected mice. These data demonstrate that NK cells have the strain specific recognition capacity required to meditate within-host competition between strains of MCMV. Furthermore, this within-host competition has the capacity to shape the dynamics of viral shedding and potentially select for the transmission of more virulent virus strains.


Estimating Learning Effects: A Short-Time Fourier Transform Regression Model for MEG Source Localization.
Magnetoencephalography (MEG) has a high temporal resolution well-suited for studying perceptual learning. However, to identify where learning happens in the brain, one needs to apply source localization techniques to project MEG sensor data into brain space. Previous source localization methods, such as the short-time Fourier transform (STFT) method by Gramfort et al.([6]) produced intriguing results, but they were not designed to incorporate trial-by-trial learning effects. Here we modify the approach in [6] to produce an STFT-based source localization method (STFT-R) that includes an additional regression of the STFT components on covariates such as the behavioral learning curve. We also exploit a hierarchical L21 penalty to induce structured sparsity of STFT components and to emphasize signals from regions of interest (ROIs) that are selected according to prior knowledge. In reconstructing the ROI source signals from simulated data, STFT-R achieved smaller errors than a two-step method using the popular minimum-norm estimate (MNE), and in a real-world human learning experiment, STFT-R yielded more interpretable results about what time-frequency components of the ROI signals were correlated with learning.


Back to the future: inhaled drug products.
Inhaled therapeutic aerosols continue to be an important treatment for asthma and pulmonary diseases. A variety of dosage forms are employed for different indications and demographics including pressurized or propellant-driven metered dose inhalers, dry powder inhalers, and nebulizers/nebules. Research and development in this field has shown remarkable innovation in the past decade. Important new drug products for the treatment of asthma, chronic obstructive pulmonary disease, cystic fibrosis, diabetes, and a range of neurological disorders have been developed. New devices in each of the dosage form categories also have been developed, and new formulation technologies have been adopted. Unlike many other dosage forms, as new inhaled products appeared few of the existing products were converted to generic form. This may be explained by the formulation and device complexity, the implications for degree of difficulty in obtaining regulatory approval, and the prevalence of intellectual property in the field. After the setback of the initial approval and subsequent withdrawal of the Exubera®-inhaled insulin, there appeared to be reluctance to consider the pulmonary route of administration for systemically acting agents, particularly peptides and proteins. However, recent product development activities and approvals suggest that attitudes may be changing in favor of systemic delivery following inhaled aerosol administration. The new inhaled drug technologies seem to be driving reconsideration of therapeutic categories for indications that were of interest at the inception of modern inhaled drug therapy in the past century. We should embrace the opportunity to use new drugs and technologies to go back to the future!


Phenotypic and genetic divergence among harbour porpoise populations associated with habitat regions in the North Sea and adjacent seas.
Determining the mechanisms that generate population structure is essential to the understanding of speciation and the evolution of biodiversity. Here, we investigate a geographical range that transects two habitat gradients, the North Sea to North Atlantic transition, and the temperate to subpolar regions. We studied the harbour porpoise (Phocoena phocoena), a small odontocete inhabiting both subpolar and temperate waters. To assess differentiation among putative populations, we measured morphological variation at cranial traits (N = 462 individuals) and variation at eight microsatellite loci for 338 of the same individuals from Norwegian, British and Danish waters. Significant morphological differentiation reflected the size of the buccal cavity. Porpoises forage in relatively shallow waters preying mainly on benthic species in British and Danish waters, and on mesopelagic and pelagic fish off the coast of Norway. We suggest that the observed differentiation may be explained by resource specialization and either adaptation or developmental responses to different local habitats.


Effect-based trigger values for in vitro and in vivo bioassays performed on surface water extracts supporting the environmental quality standards (EQS) of the European Water Framework Directive.
Effect-based methods including cell-based bioassays, reporter gene assays and whole-organism assays have been applied for decades in water quality monitoring and testing of enriched solid-phase extracts. There is no common EU-wide agreement on what level of bioassay response in water extracts is acceptable. At present, bioassay results are only benchmarked against each other but not against a consented measure of chemical water quality. The EU environmental quality standards (EQS) differentiate between acceptable and unacceptable surface water concentrations for individual chemicals but cannot capture the thousands of chemicals in water and their biological action as mixtures. We developed a method that reads across from existing EQS and includes additional mixture considerations with the goal that the derived effect-based trigger values (EBT) indicate acceptable risk for complex mixtures as they occur in surface water. Advantages and limitations of various approaches to read across from EQS are discussed and distilled to an algorithm that translates EQS into their corresponding bioanalytical equivalent concentrations (BEQ). The proposed EBT derivation method was applied to 48 in vitro bioassays with 32 of them having sufficient information to yield preliminary EBTs. To assess the practicability and robustness of the proposed approach, we compared the tentative EBTs with observed environmental effects. The proposed method only gives guidance on how to derive EBTs but does not propose final EBTs for implementation. The EBTs for some bioassays such as those for estrogenicity are already mature and could be implemented into regulation in the near future, while for others it will still take a few iterations until we can be confident of the power of the proposed EBTs to differentiate good from poor water quality with respect to chemical contamination.


In situ speciation of dissolved inorganic antimony in surface waters and sediment porewaters: development of a thiol-based diffusive gradients in thin films technique for Sb(III).
Antimony is a priority environmental contaminant typically present as either the trivalent (Sb(III)) or the pentavalent (Sb(V)) oxidation state in aquatic systems. Both the toxicity and mobility of antimony are affected by its speciation, and thus the accurate measurement of antimony speciation is essential for investigating the behaviour of this contaminant in aquatic systems. Here we present a diffusive gradients in thin films (DGT) technique, which utilises a binding layer containing a thiol-based adsorbent (3-mercaptopropyl functionalised silica gel), for the selective measurement of Sb(III) in surface waters and sediment porewaters. We also evaluated the Metsorb DGT technique, which has been previously reported to accurately measure Sb(V), for its ability to accumulate Sb(III) and thus allow the measurement of total inorganic antimony. Both the mercapto-silica and Metsorb DGT techniques showed a high affinity for Sb(III), with uptake efficiencies >97%. Elution efficiencies of 86.9 ± 2.6% and 88.1 ± 1.2% were obtained for mercapto-silica and Metsorb, respectively, with 1 mol L(-1) H2O2 in 1 mol L(-1) NaOH. The accumulation of Sb(III) by these DGT techniques was linear with time (R(2) > 0.99) and unaffected by pH (4.07-8.05), ionic strength (0.001-1.0 mol L(-1) NaCl), bicarbonate (1-15 mmol L(-1)), and an artificial seawater matrix (pH 8.34; salinity 34.8). Finally, the mercapto-silica DGT technique was applied to measure porewater concentrations of Sb(III) and As(III) in a contaminated freshwater sediment at high resolution.


Atrial fibrillation in Mediterranean spotted fever.
Mediterranean spotted fever (MSF) is a tick-borne acute febrile disease caused by Rickettsia conorii and characterized by fever, maculo-papular rash and a black eschar at the site of the tick bite ('tache noir'). We describe the case of a 58-year-old man affected by MSF who developed atrial fibrillation. The patient presented himself to the hospital after 7 days of fever, malaise and severe headache. Cardiac auscultation revealed a chaotic heart rhythm and an electrocardiogram confirmed atrial fibrillation with a fast ventricular response. Diagnosis of MSF was made after the appearance of a maculo-papular skin rash, and treatment with oral doxycycline was started. An immunofluorescence antibody test confirmed R. conorii infection. The patient recovered after 7 days of treatment. Cardiac arrhythmia is a rare complication of MSF. Inflammation may play a role in the pathogenesis of atrial fibrillation. R. conorii is an intracellular bacterium which could trigger atrial fibrillation. Our patient was previously healthy and had no reported history of cardiac disease. This suggests that heart function should be monitored in MSF patients even in the absence of underlying risk factors.


Extracellular Matrix Modulation Is Driven by Experience-Dependent Plasticity During Stroke Recovery.
Following stroke, complete cellular death in the ischemic brain area may ensue, with remaining brain areas undergoing tissue remodelling to various degrees. Experience-dependent brain plasticity exerted through an enriched environment (EE) promotes remodelling after central nervous system injury, such as stroke. Post-stroke tissue reorganization is modulated by growth inhibitory molecules differentially expressed within the ischemic hemisphere, like chondroitin sulfate proteoglycans found in perineuronal nets (PNNs). PNNs in the neocortex predominantly enwrap parvalbumin-containing GABAergic (PV/GABA) neurons, important in sensori-information processing. Here, we investigate how extracellular matrix (ECM) proteases and their inhibitors may participate in the regulation of PNN integrity during stroke recovery. Rats were subjected to photothrombotic stroke in the motor cortex, and functional deficits were assessed at 7 days of recovery. Sham and stroked rats were housed in either standard or EE conditions for 5 days, and infarct volumes were calculated. PNNs were visualized by immunohistochemistry and counted in the somatosensory cortex of both hemispheres. mRNA expression levels of ECM proteases and protease inhibitors were assessed by RT-qPCR and their activity analyzed by gel zymography. PNNs and protease activity were also studied in brains from stroke patients where similar results were observed. EE starting 2 days after stroke and continuing for 5 days stimulated behavioral recovery of limb-placement ability without affecting infarct size. EE promoted a decrease of PNNs around PV/GABA neurons and a concomitant modulation of the proteolytic activity and mRNA expression of ECM proteases and protease inhibitors in the somatosensory cortex. This study provides molecular targets for novel therapies that could support rehabilitation of stroke patients.


Increased radiosensitivity in cells of two human cell lines treated with bystander medium from irradiated repair-deficient cells.
Radiation-induced bystander factors have been shown to be more toxic if they are from medium harvested from irradiated repair-deficient cells. The aim of this study was to test the hypothesis that the radiosensitivity of repair-proficient cells can be increased by exposing them to medium-borne factors harvested from sensitive cells and vice versa. Cells from a mismatch repair (MMR)-deficient cell line (Raji 10) with a sensitive response to radiation or the wild-type parent cell line were irradiated to 0.5 Gy gamma rays and then monitored for growth rate in their own medium or in the alternative conditioned medium. In other experiments, cells or conditioned medium were added to reporter cells (HPV-G, which are relatively sensitive keratinocytes, or highly radioresistant HT29 cells). The subsequent responses of the two cell lines to a 0.5-Gy dose of (60)Co gamma rays were measured. The results show that prior exposure of resistant cells to medium from irradiated sensitive cells reduced the clonogenic survival of the subsequently irradiated resistant cells. The reverse is also true. Measurement of the apoptosis index and BCL2 expression confirmed that the harvested medium was capable of modulating apoptosis after irradiation. This may have important applications in tumor therapy and also in the understanding of mechanisms involved in induction of adaptive responses.


The predictive value of prostate cancer biomarkers depends on age and time to diagnosis: towards a biologically-based screening strategy.
Both benign and malignant prostate diseases elevate total prostate-specific antigen (tPSA), and the incidence of benign disease increases markedly with age. There is evidence, however, that free-to-total PSA ratio (%fPSA) and human kallikrein 2 (hK2) more closely reflect the malignant process. We tested the hypothesis that tPSA levels are more strongly predictive of cancer in younger when compared to older men, whereas %fPSA and hK2 are more strongly predictive in men tested closer to diagnosis. The study included 13,676 men age >/= 44 in Sweden, where PSA screening was uncommon during the study period. fPSA, tPSA and hK2 were measured in archived plasma collected during 1974-1986 in 501 men subsequently diagnosed with prostate cancer up to 1999 and in 1,292 matched controls. The predictive value of tPSA was lower in older men (p = 0.003) but was not strongly affected by time to diagnosis (p = 0.3); the predictive value of hK2 was higher closer to diagnosis (p < 0.0005) but was not modified by age (p = 0.7). A model including tPSA, fPSA and hK2 was superior (p = 0.02) to tPSA alone in older (AUC 0.819 vs. 0.794), but not in younger men (0.758 vs. 0.759). Total PSA can be used as a single marker at early middle age to predict long-term risk of prostate cancer and thus to determine intensity of subsequent screening. In contrast, %fPSA and hK2 add important predictive value in older men and much closer to diagnosis. Strategies for prostate cancer screening should be based on thorough understanding of the interaction of kallikrein-related biomarkers with prostate pathobiology.


Current concepts in host-microbe interaction leading to pneumococcal pneumonia.
Infection with Streptococcus pneumoniae (pneumococcus) results in colonization, which can lead to local or invasive disease, of which pneumonia is the most common manifestation. Despite the availability of pneumococcal vaccines, pneumococcal pneumonia is the leading cause of community and inhospital pneumonia in the United States and globally. This article discusses new insights into the pathogenesis of pneumococcal disease. The host-microbe interactions that determine whether pneumococcal colonization will result in clearance or invasive disease are highly complex. This article focuses on new information in three areas that bear on the pathogenesis of pneumococcal disease: factors that govern colonization, the prelude to invasive disease, including effects on colonization and invasion of capsular serotype, pneumolysin, surface proteins that regulate complement deposition, biofilm formation and agglutination; the effect of coinfection with other bacteria and viruses on pneumococcal growth and virulence, including the synergistic effect of influenza virus; and the contribution of the host inflammatory response to the pathogenesis of pneumococcal pneumonia, including the effects of pattern recognition molecules, cells that enhance and modulate inflammation, and therapies that modulate inflammation, such as statins. Recent research on pneumococcal pathogenesis reveals new mechanisms by which microbial factors govern the ability of pneumococcus to progress from the state of colonization to disease and host inflammatory responses contribute to the development of pneumonia. These mechanisms suggest that therapies which modulate the inflammatory response could hold promise for ameliorating damage stemming from the host inflammatory response in pneumococcal disease.


3D Printing Polymers with Supramolecular Functionality for Biological Applications.
Supramolecular chemistry continues to experience widespread growth, as fine-tuned chemical structures lead to well-defined bulk materials. Previous literature described the roles of hydrogen bonding, ionic aggregation, guest/host interactions, and π-π stacking to tune mechanical, viscoelastic, and processing performance. The versatility of reversible interactions enables the more facile manufacturing of molded parts with tailored hierarchical structures such as tissue engineered scaffolds for biological applications. Recently, supramolecular polymers and additive manufacturing processes merged to provide parts with control of the molecular, macromolecular, and feature length scales. Additive manufacturing, or 3D printing, generates customizable constructs desirable for many applications, and the introduction of supramolecular interactions will potentially increase production speed, offer a tunable surface structure for controlling cell/scaffold interactions, and impart desired mechanical properties through reinforcing interlayer adhesion and introducing gradients or self-assembled structures. This review details the synthesis and characterization of supramolecular polymers suitable for additive manufacture and biomedical applications as well as the use of supramolecular polymers in additive manufacturing for drug delivery and complex tissue scaffold formation. The effect of supramolecular assembly and its dynamic behavior offers potential for controlling the anisotropy of the printed objects with exquisite geometrical control. The potential for supramolecular polymers to generate well-defined parts, hierarchical structures, and scaffolds with gradient properties/tuned surfaces provides an avenue for developing next-generation biomedical devices and tissue scaffolds.


Optimization conditions for extracting polysaccharide from Angelica sinensis and its antioxidant activities.
In this study, polysaccharides from Angelica sinensis were extracted using the ultrasound-assisted extraction method. Based on the results of single factor experiments and orthogonal tests, three independent variables-water/raw material ratio, ultrasound time, and ultrasound power-were selected for investigation. Then, we used response surface methodology to optimize the extraction conditions. The experimental data were fitted to a quadratic equation using multiple regression analysis, and the optimal conditions were as follows: water/raw material ratio, 43.31 mL/g; ultrasonic time, 28.06 minutes; power, 396.83 W. Under such conditions, the polysaccharide yield was 21.89±0.21%, which was well matched with the predicted yield. In vitro assays, scavenging activity of superoxide anion radicals, hydroxyl radicals, and 2,2-diphenyl-1-picry-hydrazyl radical showed that polysaccharides had certain antioxidant activities and that hydroxyl radicals have a remarkable scavenging capability. Therefore, these studies provide reference for further research and rational development of A. sinensis polysaccharide.


Effect of Surgical and Immunological Castration on Haematological Variables, Reproductive Hormones and Ejaculate Characteristics in Mongrel Dogs.
Welfare concerns are growing regarding surgical castration (SC) in pets, necessitating the need for non-surgical alternatives. Administration of vaccines against gonadotropins releasing hormone (GnRH) have been reported as alternative to SC. This study determined the effect of surgical and immunological castrations (IC) on complete blood counts, plasma testosterone (T), luteinizing hormone (LH) concentrations and ejaculate characteristics in mongrel dogs. Ten intact male dogs were randomly divided into two groups (A & B). Dogs in group A were surgically castrated, while dogs in group B were immunologically castrated with single subcutaneous injection of GnRH vaccine (Improvac®). Blood and semen were collected before SC or IC and fortnightly until sixteen weeks. Blood was analyzed for packed cell volume (PCV), white blood cell count (WBC), haemoglobin concentration (Hb), absolute neutrophil (NEUT) and lymphocyte counts (LYMP), T and LH. Sperm volume (SV), concentration (C), motility (SM), live-dead ratio (LDR) and percentage of abnormal spermatozoa were determined for the semen. Data were presented as mean ± standard deviation and compared using analysis of variance. The PCV and HB of dogs surgically castrated increased progressively up to16th week after castration but only up to10 weeks in dogs immunologically castrated. Both PCV and HB decreased progressively after 10 weeks in dogs immunologically castrated. Similarly, the WBC of dogs surgically castrated steadily increased from 2 weeks up to week 16, while it increased from 6 weeks up to 16 weeks in dogs immunologically castrated. However, PCV, Hb, WBC, NEUT and LYMP did not differ significantly (p > 0.05) between SC and IC. In both groups, the SV, SC, SM, LDR and percentage of abnormal spermatozoa did not differ significantly. It was therefore concluded that there is no significant haematological or endocrinological changes between surgical and immunological castration and that immunological castration may provide safer alternative.


Factors influencing the use of reproductive health care services among married adolescent girls in Dang District, Nepal: a qualitative study.
Child marriage is associated with adverse reproductive health outcomes, and the practice is still alarmingly common. Together with efforts to end child marriage, it is essential to provide adequate health care to already married adolescents. However, to date there has been very limited research on health care-seeking practices among married adolescents in Nepal. The study was conducted in a rural part of Dang District situated in the Mid-western region of Nepal. We combined thirteen individual interviews and four focus group discussions with 17-20 years old women who had married before the age of 18 years and individual interviews with 10 key informants. Pressure to give birth early, limited autonomy, and little knowledge about reproductive health issues make married adolescents vulnerable to risky pregnancies. Early-married women face a range of barriers to use existing health services including work overload, transport and distance to health care facilities, qualities of services, verbal abuse by health care providers, and shyness and embarrassment. Women who marry and become pregnant during adolescence face a number of barriers that limit their access to health care services and they need more attention from the health services and policy makers. More youth friendly health services and education about sexual and reproductive health should be key elements in strategies to address the health issues of early-married women and adolescent girls.


Study newsletters, community and ethics advisory boards, and focus group discussions provide ongoing feedback for a large biobank.
The Personalized Medicine Research Project (PMRP) is a population-based biobank with more than 20,000 adult participants in central Wisconsin. A Community Advisory Group (CAG) and Ethics and Security Advisory Board (ESAB) provide ongoing feedback. In addition, the study newsletter is used as a two-way communication tool with study participants. The aim of this study was to assess and compare feedback received from these communication/consultation strategies with results from focus group discussions in relation to protocol changes. In summer 2009, enrollee focus groups were held addressing these topics: newsletter format, readability, and content of three articles written to solicit PMRP subject feedback. The CAG and ESAB jointly reviewed focus group results, discussed protocol changes to access residual blood samples, and made recommendations about the general communication approach. Nearly everyone in three focus groups stated that they wanted more information about PMRP. No focus group participant said that accessing stored samples would have changed their enrollment decision. Most said they wanted to be informed directly about changes affecting their original consent. For minimal-risk PMRP protocol changes, the community, CAG, and ESAB were comfortable with an opt-out model because of the initial broad consent. The planned duration of the biobank extends for decades; therefore regular, ongoing communication to enrollees is necessary to maintain awareness and trust, especially relating to protocol changes reflecting evolving science. The multi-faceted approach to communication including newsletters, external advisory boards, and focus group discussions has been successful for the PMRP biobank and may be a model for others to consider.


Bone marrow fibrosis in myelofibrosis: pathogenesis, prognosis and targeted strategies.
Bone marrow fibrosis is a central pathological feature and World Health Organization major diagnostic criterion of myelofibrosis. Although bone marrow fibrosis is seen in a variety of malignant and non-malignant disease states, the deposition of reticulin and collagen fibrosis in the bone marrow of patients with myelofibrosis is believed to be mediated by the myelofibrosis hematopoietic stem/progenitor cell, contributing to an impaired microenvironment favoring malignant over normal hematopoiesis. Increased expression of inflammatory cytokines, lysyl oxidase, transforming growth factor-β, impaired megakaryocyte function, and aberrant JAK-STAT signaling have all been implicated in the pathogenesis of bone marrow fibrosis. A number of studies indicate that bone marrow fibrosis is an adverse prognostic variable in myeloproliferative neoplasms. However, modern myelofibrosis prognostication systems utilized in risk-adapted treatment approaches do not include bone marrow fibrosis as a prognostic variable. The specific effect on bone marrow fibrosis of JAK2 inhibition, and other rationally based therapies currently being evaluated in myelofibrosis, has yet to be fully elucidated. Hematopoietic stem cell transplantation remains the only curative therapeutic approach that reliably results in resolution of bone marrow fibrosis in patients with myelofibrosis. Here we review the pathogenesis, biological consequences, and prognostic impact of bone marrow fibrosis. We discuss the rationale of various anti-fibrogenic treatment strategies targeting the clonal hematopoietic stem/progenitor cell, aberrant signaling pathways, fibrogenic cytokines, and the tumor microenvironment.


Antiplasmodial prenylated flavanonols from Tephrosia subtriflora.
The CH2Cl2/MeOH (1:1) extract of the aerial parts of Tephrosia subtriflora afforded a new flavanonol, named subtriflavanonol (1), along with the known flavanone spinoflavanone B, and the known flavanonols MS-II (2) and mundulinol. The structures were elucidated by the use of NMR spectroscopy and mass spectrometry. The absolute configuration of the flavanonols was determined based on quantum chemical ECD calculations. In the antiplasmodial assay, compound 2 showed the highest activity against chloroquine-sensitive Plasmodium falciparum reference clones (D6 and 3D7), artemisinin-sensitive isolate (F32-TEM) as well as field isolate (KSM 009) with IC50 values 1.4-4.6 μM without significant cytotoxicity against Vero and HEp2 cell lines (IC50 > 100 μM). The new compound (1) showed weak antiplasmodial activity, IC50 12.5-24.2 μM, but also showed selective anticancer activity against HEp2 cell line (CC50 16.9 μM).


Synthesis of the 2-methylene analogue of the HRV 3C protease inhibitor thysanone (2-carbathysanone).
The Human Rhinovirus (HRV) is the major aetiological agent for the common cold, for which only symptomatic treatment is available. HRV maturation and replication is entirely dependent on the activity of a virally encoded 3C protease that represents an attractive target for the development of therapeutics to treat the common cold. Herein we report the synthesis and biological evaluation of the 2-methylene analogue of the HRV 3C protease inhibitor (-)-thysanone (1) namely 2-carbathysanone (2), in an attempt to decipher the structural features in the natural product that are responsible for the 3C protease activity. 2-Carbathysanone (2) (and related analogues (±)-cis-23, (±)-cis-30, (±)-31) did not inhibit HRV 3C protease, indicating that the lactol functionality present in (-)-thysanone (1) is a critical structural feature required for inhibition.


Cochlear Implant Electrode Localization Using an Ultra-High Resolution Scan Mode on Conventional 64-Slice and New Generation 192-Slice Multi-Detector Computed Tomography.
A new generation 192-slice multi-detector computed tomography (MDCT) clinical scanner provides enhanced image quality and superior electrode localization over conventional MDCT. Currently, accurate and reliable cochlear implant electrode localization using conventional MDCT scanners remains elusive. Eight fresh-frozen cadaveric temporal bones were implanted with full-length cochlear implant electrodes. Specimens were subsequently scanned with conventional 64-slice and new generation 192-slice MDCT scanners utilizing ultra-high resolution modes. Additionally, all specimens were scanned with micro-CT to provide a reference criterion for electrode position. Images were reconstructed according to routine temporal bone clinical protocols. Three neuroradiologists, blinded to scanner type, reviewed images independently to assess resolution of individual electrodes, scalar localization, and severity of image artifact. Serving as the reference standard, micro-CT identified scalar crossover in one specimen; imaging of all remaining cochleae demonstrated complete scala tympani insertions. The 192-slice MDCT scanner exhibited improved resolution of individual electrodes (p < 0.01), superior scalar localization (p < 0.01), and reduced blooming artifact (p < 0.05), compared with conventional 64-slice MDCT. There was no significant difference between platforms when comparing streak or ring artifact. The new generation 192-slice MDCT scanner offers several notable advantages for cochlear implant imaging compared with conventional MDCT. This technology provides important feedback regarding electrode position and course, which may help in future optimization of surgical technique and electrode design.


A new role for Rab GTPases during early mitotic stages.
A recent study revealed new roles for the Rab11 GTPase during mitosis. Rab11 is involved in recycling endosome localization to mitotic spindle poles via dynein-mediated transport. This process is in contrast to Golgi membranes, which disperse in mitosis and do not appear to directly contribute to mitotic functions. Rab11-depletion prevents recycling endosome organization at spindle poles, delays mitotic progression, and disrupts spindle pole protein recruitment, astral microtubule organization, and mitotic spindle orientation. However, Rab11 is not the only endocytic and/or trafficking protein that regulates mitotic progression. Clathrin and two small GTPases (Rab6A', Rab5) play key roles in spindle organization and function. In this commentary, we discuss the roles of all these canonical endocytic and membrane trafficking proteins during mitosis and speculate on possible cross-communication between them and their molecular pathways that ensure faithful progression through mitosis.


Effects of ethanol (E85) versus gasoline vehicles on cancer and mortality in the United States.
Ethanol use in vehicle fuel is increasing worldwide, but the potential cancer risk and ozone-related health consequences of a large-scale conversion from gasoline to ethanol have not been examined. Here, a nested global-through-urban air pollution/weather forecast model is combined with high-resolution future emission inventories, population data, and health effects data to examine the effect of converting from gasoline to E85 on cancer, mortality, and hospitalization in the United States as a whole and Los Angeles in particular. Under the base-case emission scenario derived, which accounted for projected improvements in gasoline and E85 vehicle emission controls, it was found that E85 (85% ethanol fuel, 15% gasoline) may increase ozone-related mortality, hospitalization, and asthma by about 9% in Los Angeles and 4% in the United States as a whole relative to 100% gasoline. Ozone increases in Los Angeles and the northeast were partially offset by decreases in the southeast. E85 also increased peroxyacetyl nitrate (PAN) in the U.S. but was estimated to cause little change in cancer risk. Due to its ozone effects, future E85 may be a greater overall public health risk than gasoline. However, because of the uncertainty in future emission regulations, it can be concluded with confidence only that E85 is unlikely to improve air quality over future gasoline vehicles. Unburned ethanol emissions from E85 may result in a global-scale source of acetaldehyde larger than that of direct emissions.


Features of serial cranial ultrasound detected neuropathology in very preterm infants.
The purpose of this study was to identify the pattern and factors associated with changes in cranial ultrasound (CUS) -detected findings in infants born at or less than 28 weeks of gestation. We compared readings of CUS performed at the end of the first week of life and at 4-5 weeks of age. Alteration of CUS findings was classified as: (i) unchanged, if no deviation was detected (Group 1); (ii) worsening, if there were new findings (Group 2); and (iii) improvement, if there was normalization or reduction in severity (Group 3). Descriptive statistics, multivariate controlled logistic regression, and kappa (k) statistics with 95% Confidence Interval (95% CI) were reported. Among 510 studied infants, 82.3% (95% CI 78.8-85.4) were in Group 1, 10.0% (95% CI 7.7-12.9) in Group 2, and 7.7% (95% CI 5.7-10.3) in Group 3. Overall agreement between the two scans was moderate (k 0.62; 95% CI 0.55-0.69). Worsening of CUS findings was associated with neonatal morbidities independently from gestational age and birth weight. The probability for worsening of CUS findings was higher in infants with an initial diagnosis of intraventricular hemorrhage (IVH) grade 2, than in those reported as no pathology/IVH grade 1 (Odds Ratio 5.79; 95% CI 2.42-13.91) or IVH grade 3-4 (Odds Ratio 3.81; 95% CI 1.10-13.21). In very preterm born infants, the initial CUS findings in combination with neonatal morbidities can help predict the brain lesions that are seen at the end of the first month of life and could be useful in their clinical management.


Whole gut microbiome composition of damselfish and cardinalfish before and after reef settlement.
The Pomacentridae (damselfish) and Apogonidae (cardinalfish) are among the most common fish families on coral reefs and in the aquarium trade. Members of both families undergo a pelagic larvae phase prior to settlement on the reef, where adults play key roles in benthic habitat structuring and trophic interactions. Fish-associated microbial communities (microbiomes) significantly influence fish health and ecology, yet little is known of how microbiomes change with life stage. We quantified the taxonomic (16S rRNA gene) composition of whole gut microbiomes from ten species of damselfish and two species of cardinalfish from Lizard Island, Australia, focusing specifically on comparisons between pelagic larvae prior to settlement on the reef versus post-settlement juvenile and adult individuals. On average, microbiome phylogenetic diversity increased from pre- to post-settlement, and was unrelated to the microbial composition in the surrounding water column. However, this trend varied among species, suggesting stochasticity in fish microbiome assembly. Pre-settlement fish were enriched with bacteria of the Endozoicomonaceae, Shewanellaceae, and Fusobacteriaceae, whereas settled fish harbored higher abundances of Vibrionaceae and Pasteurellaceae. Several individual operational taxonomic units, including ones related to Vibrio harveyi, Shewanella sp., and uncultured Endozoicomonas bacteria, were shared between both pre and post-settlement stages and may be of central importance in the intestinal niche across development. Richness of the core microbiome shared among pre-settlement fish was comparable to that of settled individuals, suggesting that changes in diversity with adulthood are due to the acquisition or loss of host-specific microbes. These results identify a key transition in microbiome structure across host life stage, suggesting changes in the functional contribution of microbiomes over development in two ecologically dominant reef fish families.


Clinical Outcome of Probing in Infants with Acute Dacryocystitis - A Prospective Study.
Acute dacryocystitis is an uncommon but serious condition in infants and needs immediate treatment. Although, medical management with systemic antibiotics remains the mainstay of initial treatment, there are recent studies justifying simultaneous Naso Lacrimal Duct (NLD) probing with acceptable success rate. To assess the success rate of probing in the management of infantile acute dacryocystitis and to analyse the factors affecting it. A prospective interventional study was designed and infants presenting with acute dacryocystitis with or without complications like dacryocystopyocele/ lacrimal abscess/preseptal cellulitis at a Tertiary Eye Care Centre from May 2014 to April 2016 were enrolled. Demographic details and baseline clinical characteristics were noted. Intravenous antibiotics were started and probing under general anaesthesia was done. Follow up was done after one month. Success was defined as subsidence of acute attack and resolution of epiphora at final follow up of one month. A total 20 eyes of 18 patients were included with male: female ratio 10:8. Mean age of patients was 6.5 months and the mean duration of symptoms was 5.6 days. Dacryocystitis with lacrimal swelling was present in five eyes, preseptal cellulitis was present in two eyes while the rest 13 eyes presented with simple inflammation over the lacrimal sac. After one month, 85% (17eyes) had complete resolution of symptoms. Recurrence of epiphora was seen in 15% (three eyes) after initial improvement. Medical management with simultaneous probing of nasolacrimal duct has fairly good success rate in infantile dacryocystitis. Presence of dacryopyocele or dacryocystocele can lead to failure of probing owing to presence of intranasal cysts.


Application of scanning thermal microscopy for investigation of thermal boundaries in multilayered photonic structures.
In current work the application of modified Scanning Thermal Microscopy (SThM) technique for thermal imaging of multilayered periodic photonic structures is presented. The measurements were carried out using non-standard operation mode of the SThM. The thermal probe was driven by the sum of DC and small AC currents. The main advantages of presented approach are mechanical stability of the probe during measurements and high sensitivity of AC signal detection by the use of lock-in amplifier. The amplitude and phase components of probe response signal are used for visualization and analysis of the thermal properties of the layer interfaces. Basing on topographic and thermal signals the thermal boundaries between layers were revealed and the periodicity of the structure was analyzed. Presented experiment indicates that the proposed method provides spatial resolution at least about 30% better than 100 nm, which is considered for standard nanofabricated thermal probes. Therefore, proposed technique may be successfully used for the thermal boundaries mapping, as well as for the high-resolution nanoscale imaging of thermal properties distribution. The results prove that thermal imaging provides additional information to that obtained by standard AFM imaging.


Anaerobic threshold assessment through the ventilatory method during roller-ski skating testing: right or wrong?
This study aimed at questioning the validity of the ventilatory method to determine the anaerobic threshold (respiratory compensation point [RCP]) during an incremental roller-ski skating test to exhaustion. Nine elite crosscountry skiers were evaluated. The skiers carried out an incremental roller-ski test on a treadmill with the V2 skating technique. Ventilatory parameters were continuously collected breath by breath, thanks to a portable gas exchange measurement system. Poling signal was obtained using instrumented ski poles. For each stage, ventilatory and poling signals were synchronized and averaged. The poor coefficient of interobserver reliability for the time at RCP confirmed the great difficulty felt by the 3 blinded reviewers for the RCP determination. Moreover, the reviewer agreed with the impossibility of determining RCP in 4 of the 9 skiers. There was no significant difference between breathing frequency (Bf) and poling frequency (Pf) during the last 8 stages. However, it seems that the differences observed during the first stages arose from the use of either a strictly 1:1 or a 1:2 Bf to Pf ratio when the exercise intensity was still moderate. So, even if there were significant differences between the frequencies, the Bf was strictly subordinate to the Pf during the entire test. In the same way, the normalized tidal volume and peak poling forces curves were superposable. These findings showed that when the upper body is mainly involved in the propulsion, the determinants of the ventilation are strictly dependent on the poling pattern during an incremental test to exhaustion. Thus, during roller-ski skating, the determination of RCP must be used cautiously because too much depending on mechanical factors.


Identification and characterization of a supraclavicular brown adipose tissue in mice.
A fundamental challenge to our understanding of brown adipose tissue (BAT) is the lack of an animal model that faithfully represents human BAT. Such a model is essential for direct assessment of the function and therapeutic potential of BAT depots in humans. In human adults, most of the thermoactive BAT depots are located in the supraclavicular region of the neck, while mouse studies focus on depots located in the interscapular region of the torso. We recently discovered BAT depots that are located in a region analogous to that of human supraclavicular BAT (scBAT). Here, we report that the mouse scBAT depot has morphological characteristics of classical BAT, possesses the potential for high thermogenic activity, and expresses a gene signature that is similar to that of human scBAT. Taken together, our studies reveal a mouse BAT depot that represents human BAT and provides a unique tool for developing new translatable approaches for utilizing human scBAT.


The added effect of comorbidity on health-related quality of life in patients with asthma.
To examine the effect of comorbidities on health-related quality of life (HRQoL) and their interaction with asthma control. In a random sample of adults with asthma, we measured generic (EQ5D) and disease-specific (AQ5D) utility scores. Asthma symptom control was determined using the 2014 Global Initiative for Asthma Management Strategy. Comorbidity scores were calculated using a validated questionnaire. We used two-part regression models to measure the adjusted difference in utility across levels of symptom control and comorbidity scores and to examine the relative role of symptom control and comorbidity in explaining the variation in HRQoL. A total of 2,299 observations from 460 adult patients (mean age 52 years, 67 % women) were included. Compared to controlled asthma, uncontrolled asthma was associated with -0.018 reduction (95 % CI -0.028, -0.009) in EQ5D and -0.076 reduction (95 % CI -0.115, -0.052) in AQ5D utilities. An increase by one standard deviation in comorbidity score relative to the mean was associated with a change of -0.029 (95 % CI -0.043, -0.016) in EQ5D and -0.010 (95 % CI -0.020, -0.004) in AQLQ utilities. The impact of comorbidity was greater than asthma symptom control in explaining EQ5D variance (12 vs. 1 %) but smaller in explaining AQ5D variance (3 vs. 12 %). Generic and disease-specific HRQoL instruments differentially capture the impact of symptom control and comorbidity in asthma. The selection of HRQoL instruments for asthma studies should depend on the prevalence of comorbidity in the target population and the impact of interventions on asthma control and comorbidity.


School holidays and admissions with asthma.
The admission rate for asthma at a children's hospital was studied over an 11 year period. Admissions varied unpredictably over periods of a few days, but there was a repeated yearly pattern of peaks and troughs with an interval of several weeks. The short term variation could be attributed to chance effects alone, excluding any important role for short term influences--for example, weather changes--in precipitating asthma admissions. There was a definite association between the longer term variation and school holidays. The admission rate fell during holidays and there were two or more peaks during terms. The pattern is consistent with a largely viral aetiology for asthmatic attacks throughout the year. We postulate that school holidays disrupt the spread of viral infections in a community, with synchronisation of subsequent attacks. Travel during holidays may facilitate acquisition of new viral strains by the community.


Publication of NIH funded trials registered in ClinicalTrials.gov: cross sectional analysis.
To review patterns of publication of clinical trials funded by US National Institutes of Health (NIH) in peer reviewed biomedical journals indexed by Medline. Cross sectional analysis. Clinical trials funded by NIH and registered within ClinicalTrials.gov (clinicaltrials.gov), a trial registry and results database maintained by the US National Library of Medicine, after 30 September 2005 and updated as having been completed by 31 December 2008, allowing at least 30 months for publication after completion of the trial. Publication and time to publication in the biomedical literature, as determined through Medline searches, the last of which was performed in June 2011. Among 635 clinical trials completed by 31 December 2008, 294 (46%) were published in a peer reviewed biomedical journal, indexed by Medline, within 30 months of trial completion. The median period of follow-up after trial completion was 51 months (25th-75th centiles 40-68 months), and 432 (68%) were published overall. Among published trials, the median time to publication was 23 months (14-36 months). Trials completed in either 2007 or 2008 were more likely to be published within 30 months of study completion compared with trials completed before 2007 (54% (196/366) v 36% (98/269); P<0.001). Despite recent improvement in timely publication, fewer than half of trials funded by NIH are published in a peer reviewed biomedical journal indexed by Medline within 30 months of trial completion. Moreover, after a median of 51 months after trial completion, a third of trials remained unpublished.


Pyle metaphyseal dysplasia in an African child: Case report and review of the literature.
Pyle disease (OMIM 265900), also known as metaphyseal dysplasia, is a rare autosomal recessive disorder with no known gene mutation. We report a case of Pyle disease in a 7-year-old African boy of mixed ancestry who presented with finger and wrist fractures following minor trauma. The radiological findings revealed abnormally broad metaphyses of the tubular bones, known as Erlenmeyer-flask bone deformity, and mild cranial sclerosis, both hallmarks of the condition. We report the first case in a patient with African ancestry, which could help in the gene discovery of this rare autosomal recessive skeletal dysplasia with unknown mutations.


In vitro processing of tomato proteinase inhibitor I by barley microsomal membranes: a system for analysis of cotranslational processing of plant endomembrane proteins.
A plant-derived in vitro system for the study of cotranslational processing of plant endomembrane proteins has been developed and used to investigate cotranslational proteolytic processing of tomato proteinase inhibitor I. Translation of the inhibitor I precursor in wheat germ lysate supplemented with barley aleurone microsomal membranes resulted in cotranslational import of the protein into microsomal vesicles and cleavage of the signal sequence. NH(2)-terminal sequence analysis of the translocated inhibitor I processing intermediate showed that the signal sequence was cleaved between Ala(23) and Arg(24) of the precursor protein. Parallel experiments using dog pancreas microsomal membranes indicated an identical site of cleavage, suggesting that the substrate determinants for signal sequence processing are conserved across kingdoms. The plant-derived processing system used for this study may be valuable for analysis of cotranslational processing of other plant preproteins and for characterizing the components of the cotranslational import machinery in plants.


Factors determining clinical outcomes in intussusception in the developing world: Experience from Johannesburg, South Africa.
Rates of open reduction of intussusception were noted to be unacceptably high during an institutional internal audit. To determine the impact of revised protocols to better select patients for pneumatic reduction (PR), and document associated morbidity and mortality, and the factors that affect the above. Medical records of patients between 3 months and 3 years of age presenting to the Department of Paediatric Surgery at Chris Hani Baragwanath Academic Hospital, Johannesburg, South Africa, from 2007 to 2010 were reviewed. Determining factors, including duration of symptoms, admission C-reactive protein (CRP) level and weight, were analysed against clinical outcomes, notably PR, bowel resection, relook laparotomy and death. A total of 97 cases were suitable for inclusion. In 62 of these (63.9%), PR was attempted; this was successful in 32 cases (51.6%), giving an overall successful PR rate of 33.0%. In 7 of the 62 patients, a pneumoperitoneum was documented during the reduction attempt. Of the 65 patients who underwent surgery, 53 required intestinal resection and 12 had spontaneous or manual reduction. Ileostomy was necessary in 9 patients, and 7 required relook laparotomy. The overall mortality rate was 9.1%. Averages of 'determining factors' assessed against clinical outcome were as follows: mean weight (standard deviation (SD)) 7.4 (4.3) kg, mean duration of symptoms (DOS) 3.0 (SD 2.2) days, and admission CRP level 50.9 mg/L (range 1 - 249.3). Prolonged DOS and a raised CRP level predicted a poor outcome. Despite marked improvements in management and PR outcomes, intussusception remains associated with significant morbidity and mortality. Prolonged DOS and an elevated CRP predict worse outcomes. The use of these markers in association with clinical factors may assist management decisions, specifically with regard to operative or non-operative management. Awareness and education are key to prompt presentation and early diagnosis. Well-defined protocols introduced at all points of contact ensure early recognition and resuscitation as well as prompt referral for definitive management.


Long-term assessment of ecosystem services at ecological restoration sites using Landsat time series.
Reversing ecological degradation through restoration activities is a key societal challenge of the upcoming decade. However, lack of evidence on the effectiveness of restoration interventions leads to inconsistent, delayed, or poorly informed statements of success, hindering the wise allocation of resources, representing a missed opportunity to learn from previous experiences. This study contributes to a better understanding of spatial and temporal dynamics of ecosystem services at ecological restoration sites. We developed a method using Landsat satellite images combined with a Before-After-Control-Impact (BACI) design, and applied this to an arid rural landscape, the Baviaanskloof in South Africa. Since 1990, various restoration projects have been implemented to halt and reverse degradation. We applied the BACI approach at pixel-level comparing the conditions of each intervened pixel (impact) with 20 similar control pixels. By evaluating the conditions before and after the restoration intervention, we assessed the effectiveness of long-term restoration interventions distinguishing their impact from environmental temporal changes. The BACI approach was implemented with Landsat images that cover a 30-year period at a spatial resolution of 30 meter. We evaluated the impact of three interventions (revegetation, livestock exclusion, and the combination of both) on three ecosystem services; forage provision, erosion prevention, and presence of iconic vegetation. We also evaluated whether terrain characteristics could partially explain the variation in impact of interventions. The resulting maps showed spatial patterns of positive and negative effects of interventions on ecosystem services. Intervention effectiveness differed across vegetation conditions, terrain aspect, and soil parent material. Our method allows for spatially explicit quantification of the long-term restoration impact on ecosystem service supply, and for the detailed visualization of impact across an area. This pixel-level analysis is specifically suited for heterogeneous landscapes, where restoration impact not only varies between but also within restoration sites.


Asymmetric catalysis activated by visible light.
Visible light driven organic chemistry has sparked much excitement over the last several years. This review summarizes recent progress in combining visible light activation with asymmetric catalysis, processes that are either mediated by photoinduced electron or energy transfer. The tasks of photoactivation and asymmetric catalysis are typically accomplished by dual catalyst systems but several recent reports demonstrate that they can also be effectively executed by single catalysts. Beyond the discovery of novel asymmetric transformations under mild reaction conditions, this contemporary area of organic chemistry holds promise for the development of economical and environmentally friendly methods for the asymmetric synthesis of chiral compounds.


Expression sequence tag library derived from peripheral blood mononuclear cells of the chlorocebus sabaeus.
African Green Monkeys (AGM) are amongst the most frequently used nonhuman primate models in clinical and biomedical research, nevertheless only few genomic resources exist for this species. Such information would be essential for the development of dedicated new generation technologies in fundamental and pre-clinical research using this model, and would deliver new insights into primate evolution. We have exhaustively sequenced an Expression Sequence Tag (EST) library made from a pool of Peripheral Blood Mononuclear Cells from sixteen Chlorocebus sabaeus monkeys. Twelve of them were infected with the Simian Immunodeficiency Virus. The mononuclear cells were or not stimulated in vitro with Concanavalin A, with lipopolysacharrides, or through mixed lymphocyte reaction in order to generate a representative and broad library of expressed sequences in immune cells. We report here 37,787 sequences, which were assembled into 14,410 contigs representing an estimated 12% of the C. sabaeus transcriptome. Using data from primate genome databases, 9,029 assembled sequences from C. sabaeus could be annotated. Sequences have been systematically aligned with ten cDNA references of primate species including Homo sapiens, Pan troglodytes, and Macaca mulatta to identify ortholog transcripts. For 506 transcripts, sequences were quasi-complete. In addition, 6,576 transcript fragments are potentially specific to the C. sabaeus or corresponding to not yet described primate genes. The EST library we provide here will prove useful in gene annotation efforts for future sequencing of the African Green Monkey genomes. Furthermore, this library, which particularly well represents immunological and hematological gene expression, will be an important resource for the comparative analysis of gene expression in clinically relevant nonhuman primate and human research.


Environmental controls over isoprene emission in deciduous oak canopies.
In summer 1992, isoprene emission was measured on intact leaves and branches of Quercus alba (L.) at two heights in a forest canopy. Isoprene emission capacity (measured at 30 degrees C and a photosynthetic photon flux density of 1000 micro mol m(-2) s(-1)) was significantly higher in sun leaves than in shade leaves when expressed on a leaf area basis (51 versus 31 nmol m(-2) s(-1); P < 0.01). Because leaf mass per unit area (LMA, g m(-2)) was higher in sun leaves than in shade leaves, emissions of sun and shade leaves expressed on a dry mass basis did not differ significantly (99 versus 89 micro g C g(DW) (-1) h(-1); P = 0.05). Similar measurements in 1995 were consistent with the 1992 data, but data from leaves in more shaded locations demonstrated that isoprene emission capacity decreased with decreasing growth irradiance, irrespective of units of expression. Isoprene emission capacity in leaves of Q. coccinea Muenchh. and Q. velutina Lam. also declined steeply with canopy depth. Emission capacity, on a dry mass basis, showed no obvious pattern with canopy position in Q. prinus L. There was no difference in the temperature response of sun versus shade leaves of Q. alba, but shade leaves exhibited a greater quantum efficiency and saturated at lower irradiance than sun leaves. Rates of isoprene emission measured on branches of Q. alba were approximately 60% of those measured on individual leaves, as a result of self-shading within branch enclosures. It is recommended that within-canopy variation in isoprene emission capacity be incorporated into regional emission models.


A novel mechanism of immune regulation: interferon-gamma regulates retention of CD4 T cells during delayed type hypersensitivity.
The local immune response is characterized by an increase in the rate of entry of lymphocytes from the blood into regional lymph nodes and changes in the output of cells in lymph. While significant data are available regarding the role of inflammation-induced vascular adhesion processes in regulating lymphocyte entry into inflamed tissues and lymph nodes, relatively little is known about the molecular processes governing lymphocyte exit into efferent lymph. We have defined a novel role for lymphatic endothelial cells in the regulation of lymphocyte exit during a delayed type hypersensitivity (DTH) response to mycobacterial purified protein derivative (PPD). Soluble, pro-adhesive factors were identified in efferent lymph concomitant with reduced lymphocyte output in lymph, which significantly increased lymphocyte binding to lymphatic endothelial cells. While all lymphocyte subsets were retained, CD4+ T cells appeared less susceptible than others. Among a panel of cytokines in inflammatory lymph plasma, interferon (IFN)-gamma alone appeared responsible for this retention. In vitro adhesion assays using physiological levels of IFN-gamma confirmed the interaction between recirculating lymphocytes and lymphatic endothelium. These data demonstrate a new level of immune regulation, whereby the exit of recirculating lymphocytes from lymph nodes is selectively and sequentially regulated by cytokines in a manner equally as complex as lymphocyte recruitment.


Changes of AI receptive fields with sound density.
Primates engage in auditory behaviors under a broad range of signal-to-noise conditions. In this study, optimal linear receptive fields were measured in alert primate primary auditory cortex (A1) in response to stimuli that vary in spectrotemporal density. As density increased, A1 excitatory receptive fields systematically changed. Receptive field sensitivity, expressed as the expected change in firing rate after a tone pip onset, decreased by an order of magnitude. Spectral selectivity more than doubled. Inhibitory subfields, which were rarely recorded at low sound densities, emerged at higher sound densities. The ratio of excitatory to inhibitory population strength changed from 14.4:1 to 1.4:1. At low sound densities, the sound associated with the evocation of an action potential from an A1 neuron was broad in spectrum and time. At high sound densities, a spike-evoking sound was more likely to be a spectral or temporal edge and was narrower in time and frequency range. Receptive fields were used to predict responses to a novel high-noise-density stimulus. The predictions were highly correlated with the actual responses to the 2-s complex sound excerpt. The structure of prediction failures revealed that neurons with prominent inhibitory fields had relatively poor linear predictions. Further, the finding that stochastic variance is limiting in prediction even after averaging 150 repetitions means that high-fidelity representations of simple sounds in A1 must be distributed over at least hundreds of neurons. Auditory context alters A1 responses across multiple parameter spaces; this presents a challenge for reconstructing neural codes.