The current goal of treatment in irritable bowel syndrome (IBS) focuses primarily on symptom management and attempts to improve quality of life. Several treatments are at the disposal of physicians; lifestyle and dietary management, pharmacological treatments and psychological interventions are the most used and recommended. Psychological treatments have been proposed as viable alternatives or compliments to existing care models. Most forms of psychological therapies studied have been shown to be helpful in reducing symptoms and in improving the psychological component of anxiety/depression and health-related quality of life. According to current NICE/NHS guidelines, physicians should consider referral for psychological treatment in patients who do not respond to pharmacotherapy for a period of 12 months and develop a continuing symptom profile (described as refractory irritable bowel syndrome). Cognitive behavioral therapy (CBT) is the best studied treatment and seems to be the most promising therapeutic approach. However, some studies have challenged the effectiveness of this therapy for irritable bowel syndrome. One study concluded that cognitive behavioral therapy is no more effective than placebo attention control condition and another study showed that the beneficial effects wane after six months of follow-up. A review of mind/body approaches to irritable bowel syndrome has therefore suggested that alternate strategies targeting mechanisms other than thought content change might be helpful, specifically mindfulness and acceptance-based approaches. In this article we review these new psychological treatment approaches in an attempt to raise awareness of alternative treatments to gastroenterologists that treat this clinical syndrome.

The use of NMR chiral solvating agents (CSAs) for the analysis of enantiopurity has been known for decades, but has been supplanted in recent years by chromatographic enantioseparation technology. While chromatographic methods for the analysis of enantiopurity are now commonplace and easy to implement, there are still individual compounds and entire classes of analytes where enantioseparation can prove extremely difficult, notably, compounds that are chiral by virtue of very subtle differences such as isotopic substitution or small differences in alkyl chain length. NMR analysis using CSAs can often be useful for such problems, but the traditional approach to selection of an appropriate CSA and the development of an NMR-based analysis method often involves a trial-and-error approach that can be relatively slow and tedious. In this study we describe a high-throughput experimentation approach to the selection of NMR CSAs that employs automation-enabled screening of prepared libraries of CSAs in a systematic fashion. This approach affords excellent results for a standard set of enantioenriched compounds, providing a valuable comparative data set for the effectiveness of CSAs for different classes of compounds. In addition, the technique has been successfully applied to challenging pharmaceutical development problems that are not amenable to chromatographic solutions. Overall, this methodology provides a rapid and powerful approach for investigating enantiopurity that compliments and augments conventional chromatographic approaches.

Systemic lupus erythematosus (SLE) is a multisystemic disease characterized by alterations in the regulation of both cellular and humoral immune responses. B cell hyperactivity and genetic aberrations lead to formation of compliment-fixing IgG autoantibodies including anti-DNA and anti-nucleosome antibodies. Pathological T cell clones that recognize double-stranded DNA and nucleosomes further drive B cell production of DNA autoantibodies. Deposition of autoantibodies within the skin, kidney, brain, and other organ systems contributes to the pathophysiology and clinical manifestations of SLE. A growing body of experimental evidence indicates that DNA antibodies contribute to the histological changes observed in lupus nephritis. The binding of anti-DNA and other autoantibodies to basement membranes and other cellular structures within the glomerulus results in activation of compliment and recruitment of inflammatory cells into the glomerulus. The use of high-dose steroid hormones and cytotoxic agents have improved patient and renal survival, but are associated with major infection, infertility, osteoporosis, and secondary malignancies. New pharmacological approaches to the treatment of lupus nephritis will include drugs that deplete specific B cell clones involved in the synthesis of nephritogenic autoantibodies as well as the blocking of signal transduction pathways required for antigen-dependent antibody synthesis. Novel clonal-specific approaches to immunosuppression in patients with SLE offer the potential for precise targeting of the disease pathogenesis and for reducing toxic complications of treatment.

Four types of radionuclide investigations are described here: 99mTc-labeled red blood cell scintigraphy, colloid liver scintigraphy, hepatobiliary scintigraphy, and positron emission tomography with [18F]fluorodeoxyglucose. The role of nuclear imaging techniques in the diagnosis of liver diseases has changed in recent years and now compliments morphological imaging modalities by offering the unique ability to visualize function and metabolism. The studies described here are therefore rarely performed now by themselves for the delineation of secondary liver tumors. These radionuclide investigations are used principally to narrow the differential diagnosis of focal liver disease.

Direction-changing nystagmus has been seen after the ingestion of ethanol in both animals and man. The direction of the nystagmus is dependent upon the position of the head and is thus called Positional Alcohol Nystagmus (PAN). This article provides information about positionally dependent, direction-changing nystagmus in human beings after the ingestion of glycerol (1.5 g/kg). Electronystagmographic recordings were made and serum glycerol levels were repeatedly determined over a period of 7 hours. The resulting data compliment earlier work done with ethanol. These data provide support for a buoyancy hypothesis to explain positional nystagmus after ingestion of water-soluble molecules with differing specific gravities.

Screening for HIV infection can use many algorithms. When two different HIV antibody assays are used, discordant results may occur. To discriminate between HIV seroconversion, HIV variant infection and false positive reactivity, 30 consecutive subjects with two discordant HIV antibody-screening assays were extensively investigated for HIV infection. No subject had HIV seroconversion or reached HIV seropositivity criteria after a follow-up of 3 months. By contrast 36% became HIV negative by the use of both HIV screening assays. p24 Antigen, HIV-1 RNA, HIV-1 DNA, HIV-2 DNA assays and HIV isolation by sensitive culture were unable to identify HIV infection in the 30 subjects with discordant HIV screening assays. The data suggest that the use of two HIV screening assays increase false-positive HIV results without increasing clinical sensitivity. To compliment follow-up of HIV screening, early testing for HIV RNA could be useful to identify or eliminate a recent infection.

The double-contrast barium enema is a highly accurate and readily available procedure which is capable of diagnosing benign and malignant disease of the colon at an early state. The level of accuracy is a function of the radiologist's knowledge and willingness to assume complete responsibility for the patient. While reasonable results may be obtained with the high-kilovoltage single-contrast method, it is our opinion that the double contrast examination is the more sensitive procedure and therefore the technique of choice. The double-contrast examination ideally compliments the colonoscope and offers an alternative method for initial survey procedures or low cost follow-up examinations.

Nuisance cyanobacterial blooms degrade water resources through accelerated eutrophication, odor generation, and production of toxins that cause adverse effects on human health. Quick and effective methods for detecting cyanobacterial abundance in drinking water supplies are urgently needed to compliment conventional laboratory methods, which are costly and time consuming. Hyperspectral remote sensing can be an effective approach for rapid assessment of cyanobacterial blooms. Samples (n=250) were collected from five drinking water sources in central Indiana (CIN), USA, and South Australia (SA), which experience nuisance cyanobacterial blooms. In situ hyperspectral data were used to develop models by relating spectral signal with handheld fluorescence probe (YSI 6600 XLM-SV) measured phycocyanin (PC in cell/ml), a proxy pigment unique for indicating the presence of cyanobacteria. Three-band model (TBM), which is effective for chlorophyll-a estimates, was tuned to quantify cyanobacteria coupled with the PC probe measured cyanobacteria. As a comparison, two band model proposed by Simis et al. (Limnol Oceanogr, 50(11): 237-245, 2005; denoted as SM05) was paralleled to evaluate TBM model performance. Our observation revealed a high correlation between measured and estimated PC for SA dataset (R (2) =0.96; range: 534-20,200 cell/ml) and CIN dataset (R (2) =0.88; range: 1,300-44,500 cell/ml). The potential of this modeling approach for imagery data were assessed by simulated ESA/Centinel3/OLCI spectra, which also resulted in satisfactory performance with the TBM for both SA dataset (RMSE % =26.12) and CIN dataset (RMSE % =34.49). Close relationship between probe-measured PC and laboratory measured cyanobacteria biovolume was observed (R (2) =0.93, p<0.0001) for the CIN dataset, indicating a stable performance for PC probe. Based on our observation, field spectroscopic measurement coupled with PC probe measurements can provide quantitative cyanobacterial bloom information from both relatively static and flowing inland waters. Hence, it has promising implications for water resource managers to obtain information for early warning detection of cyanobacterial blooms through the close association between probe measured PC values and cyanobacterial biovolume via remote sensing modeling.

OBJECTIVE

Neovascularization is essential for tumor growth and invasion. Mounting evidence suggests that tumor cells recruit circulating endothelial progenitor cells to promote vasculogenesis to compliment tumor angiogenesis. This study examines the constitutive role of bone marrow-derived cells in this process.

METHODS

Rat glioma cells were implanted into brains of T-cell-depleted knockout mice. At various timepoints after tumor implantation, naïve bone marrow cells from ubiquitous transgenic mice expressing green fluorescent protein (GFP) were infused into these animals. The incorporation of GFP-positive cells into the vascular architecture was visualized by fluorescence confocal microscopy in conjunction with the transcription profiles of vascular endothelial growth factor (VEGF) and angiopoietin-1 and -2 (Ang-1 and Ang-2).

RESULTS

Of the cells infused, 8 days after tumor implantation, 0.49% were found exclusively sequestered in the vicinity of tumor vessels. This coincided with a decline in the expression of Ang-1 and a rise in the expression of VEGF and Ang-2. A few of these cells (0.66 of the 0.49%) localized onto the vascular wall. They resembled endothelial cells and expressed vWF.

CONCLUSIONS

The incorporation of bone marrow-derived unpurified endothelial cells into the tumor vascular bed is both time-limited and infrequent. These cells may play a supportive rather than a constitutive role in tumor neovascularization.

Women with pre-gestational diabetes have a higher risk of producing children with congenital heart defects (CHDs), caused predominantly by hyperglycemia-induced oxidative stress. In this study, we evaluated if exercise during pregnancy could mitigate oxidative stress and reduce the incidence of CHDs in the offspring of diabetic mice. Female mice were treated with streptozotocin to induce pre-gestational diabetes, then mated with healthy males to produce offspring. They were also given access to running wheels 1 week before mating and allowed to exercise voluntarily until E18.5. Heart morphology, gene expression, and oxidative stress were assessed in foetal hearts. Maternal voluntary exercise results in a significantly lower incidence of CHDs from 59.5% to 25%. Additionally, diabetes-induced defects in coronary artery and capillary morphogenesis were also lower with exercise. Myocardial cell proliferation and epithelial-mesenchymal transition at E12.5 was significantly lower with pre-gestational diabetes which was mitigated with maternal exercise. Cardiac gene expression of Notch1, Snail1, Gata4 and Cyclin D1 was significantly higher in the embryos of diabetic mice that exercised compared to the non-exercised group. Furthermore, maternal exercise produced lower reactive oxygen species (ROS) and oxidative stress in the foetal heart. In conclusion, maternal exercise mitigates ROS and oxidative damage in the foetal heart, and results in a lower incidence of CHDs in the offspring of pre-gestational diabetes. Exercise may be an effective intervention to compliment clinical management and further minimize CHD risk in mothers with diabetes.

INTRODUCTION. The impact of preformed donor-specific antibodies (DSA) is incompletely understood in liver transplantation. The incidence and impact of preformed DSA on early post liver transplant were assessed and these were correlated with compliment fragment C4d on allograft biopsy. METHODS. Pretransplant serum from 41 consecutive liver transplant recipients (brain dead donors; DBD = 27 and cardiac death donors; DCD = 14) were tested for class-specific anti-human leukocyte antigen (HLA) and compared against donor HLA types. Liver biopsies were taken during cold storage (t-1) and post-reperfusion (t0) stained with C4d and graded for preservation-reperfusion injury (PRI). RESULTS. Of the 41 recipients, 8 (20%) had anti-HLA class I/II antibodies pretransplant, 3 (7%) were confirmed preformed DSA; classes I and II (n=1) and class I only (n=2). No biopsies showed definite evidence of antibody-mediated rejection. Graft biopsies in overall showed only mild PRI with ischemic hepatocyte C4d pattern similar in both positive and negative DSA patients. One DSA-positive (33%) compared with four DSA-negative patients (10%) had significant early graft dysfunction; severe PRI causing graft loss from primary nonfunction was seen only in DSA-negative group. Allograft biopsy of preformed DSA-positive patient demonstrated only minimal PRI; however, no identifiable cause could be attributed to graft dysfunction other than preformed DSA. CONCLUSION. Preformed DSA are present in 5-10% liver transplant recipients. There is no association between anti-HLA DSA and PRI and C4d, but preformed DSA may cause early morbidity. Larger studies on the impact of DSA with optimization of C4d techniques are required.

To further characterize the toxicological risk associated with chemical contaminants in Great Lakes fish, a multigeneration rat reproduction study was designed. Mature chinook salmon (Oncorhynchus tsawytscha), collected during the Fall 1991 spawning runs from Sydenham River, Lake Huron, and Credit River, Lake Ontario, were filleted, lyophilized, and incorporated into standard rat diets at 25% (w/w) or 100% (w/w) of the normal protein compliment [casein, 20% (w/w)]. This resulted in diets composed of 5 or 20% (w/w) lyophilized fish and estimated daily fish intakes by the rats at levels approximately 15- and 60-fold greater, respectively, than the current estimate for the Canadian public for all fish and seafood. Both fresh and lyophilized fish were analyzed for the following groups of contaminants: halogenated aromatic hydrocarbons [polychlorinated biphenyls, dibenzodioxins, and dibenzofurans (PCBs, PCDDs, PCDFs)], polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides, metals, volatile organics, and other extractable organics (chlorinated phenols and benzenes). In general, only minor site differences existed for the specific types of contaminants detected; however, fish from the Credit River contained slightly greater amounts of PCBs (2- to 3-fold), dioxin toxic equivalencies (TCDD TEQs; 1.5- to 2.0-fold), DDT and metabolites (1. 5-fold), and appreciably higher amounts of mirex (15-fold). This general pattern of contaminant differences continued when the various diets were prepared using the lyophilized fish. Tissue samples (adipose, liver) were taken from the animals at various stages of the study and also analyzed for the same groups of contaminants. In general, adipose tissue was the major reservoir for organochlorine (OC) pesticides and PCBs, while "dioxin-like" PCDD/DF congeners and mercury were found preferentially in the liver. Contaminant intake calculations and tissue residue levels are provided.

Screening mammography identifies suspicious, non palpable mammary lesions. Mammographic needle localization (MNL) is currently being used to facilitate excision biopsy of these lesions. Thirty-two patients underwent biopsies of the breast after MNL for non-palpable lesions. Mammographic indications for biopsy consisted of microcalcifications (48%), mass or abnormal density (21%) or mass+abnormal density (24%). The carcinoma was identified in four cases (12%). Two of these were in situ, one was microinvasive and one was frankly invasive. Three were treated with a modified radical mastectomy. One of these non palpable lesion demonstrated nodal metastasis but none showed distant metastasis. All radiologically detected abnormalities were removed and confirmed with repeat radiology. No complications were identified. MNL effectively localizes non-palpable lesion of the breast and compliments accurate diagnosis and treatment of early carcinoma of the breast.

Cardiac dyssynchrony arises from conduction abnormalities during heart failure and worsens morbidity and mortality. Cardiac resynchronization therapy (CRT) re-coordinates contraction using bi-ventricular pacing, but the cellular and molecular mechanisms involved remain largely unknown. The aim is to determine how dyssynchronous heart failure (HFdys ) alters the phospho-proteome and how CRT interacts with this unique phospho-proteome by analyzing Ser/Thr and Tyr phosphorylation. Phospho-enriched myocardium from dog models of Control, HFdys , and CRT is analyzed via MS. There were 209 regulated phospho-sites among 1761 identified sites. Compared to Con and CRT, HFdys is hyper-phosphorylated and tyrosine phosphorylation is more likely to be involved in signaling that increased with HFdys and was exacerbated by CRT. For each regulated site, the most-likely targeting-kinase is predicted, and CK2 is highly specific for sites that are "fixed" by CRT, suggesting activation of CK2 signaling occurs in HFdys that is reversed by CRT, which is supported by western blot analysis. These data elucidate signaling networks and kinases that may be involved and deserve further study. Importantly, a possible role for CK2 modulation in CRT has been identified. This may be harnessed in the future therapeutically to compliment CRT, improving its clinical effects.

The present study was carried out to characterize Angiotensin-converting enzyme (ACE) inhibitory peptides which are released from the trypsin hydrolysate of wheat gluten protein. The binding of two inhibitory peptide (P4 and P6) to human serum albumin (HSA) under physiological conditions has been investigated by multi-spectroscopic in combination with molecular modeling techniques. Time-resolved and quenching fluorescence spectroscopies results revealed that the quenching of HSA fluorescence by P4 and P6 in the binary and ternary systems caused HSA-peptides complexes formation. The results indicated that both peptides quenched the fluorescence intensity of HSA through a static mechanism. The binding affinities and number of binding sites were obtained for the HSA-peptides complexes. The circular dichroism (CD) data revealed that the presence of both peptides increased the α-helix content of HSA and induced the remarkable folding of the polypeptide of the protein. Therefore, the CD data determined that the protein structure has been stabilized in the percent of ACE inhibitory peptides in binary and ternary systems. The binding distances between HSA and both peptides were estimated by the Forster theory, and it was revealed that nonradiative energy transfer from HSA to peptides occurred with a high probability. ITC experiments reveal that, in the absence and presence of P6, the dominant forces are electrostatic in binary and ternary systems. Furthermore, molecular modeling studies confirmed the experimental results. Molecular modeling investigation suggested that P4 bound to the site IA and IIA of HSA in binary and ternary systems, respectively. This study on the interaction of peptides with HSA should prove helpful for realizing the distribution and transportation of food compliments and drugs in vivo, elucidating the action mechanism and dynamics of food compliments and drugs at the molecular level. It should moreover be of great use for understanding the pharmacokinetic and pharmacodynamic mechanism of the food compliments and drugs.

BACKGROUND

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) remains the deadliest infectious disease in the world with one-third of the world's population thought to be infected. Over the years, TB mortality rate has been largely reduced; however, this progress has been threatened by the increasing appearance of multidrug-resistant Mtb. Considerable recent efforts have been undertaken to develop new generation antituberculosis drugs. Many of these attempts have relied on in silico approaches, which have emerged recently as powerful tools complementary to biochemical attempts. Areas covered: The authors review the status of pharmaceutical drug development against TB with a special emphasis on computational work. They focus on those studies that have been validated by in vitro and/or in vivo experiments, and thus, that can be considered as successful. The major goals of this review are to present target protein systems, to highlight how in silico efforts compliment experiments, and to aid future drug design endeavors. Expert opinion: Despite having access to all of the gene and protein sequences of Mtb, the search for new optimal treatments against this deadly pathogen are still ongoing. Together with the geometric growth of protein structural and sequence databases, computational methods have become a powerful technique accelerating the successful identification of new ligands.

The effects of trait self-objectification and compliment type (Neutral/Character/Appearance) on women's negative mood was examined. One hundred and eighty-five undergraduate women participated in mock appearance evaluations and interviews before completing questionnaires. As in previous research, women high on trait self-objectification displayed substantially greater body shame and appearance anxiety than women low on trait self-objectification. Women high on trait self-objectification who received character or appearance compliments expressed less negative mood than those receiving neutral compliments. A supplementary study (N=53) suggested that the effect of positive compliments may be due, in part, to the fact that the self-esteem of high trait self-objectified participants was largely contingent on others' approval. Offering highly trait self-objectified women positive statements concerning either their characters or their appearance temporarily assuages negative mood.

Multi-photon microscopy and advances in optics, computer sciences, and the available labeling fluorophores now allow investigators to study the dynamic events within the functioning kidney with subcellular resolution. This emerging technology, with improved spatial and temporal resolution and sensitivity, enables investigators to follow complex heterogenous processes in organs such as the kidney. Repeated determinations within the same animal are possible minimizing their use and inter-animal variability. Furthermore, the ability to obtain volumetric data (3D) makes quantitative 4D (time) analysis possible. Finally, use of up to three fluorophores concurrently allows three different or interactive processes to be observed simultaneously. Therefore, this approach compliments existing molecular, biochemical, and pharmacologic techniques by advancing data analysis and interpretation to subcellular levels for molecules without the requirement for fixation.

Background: The fungal pathogen Leptosphaeria maculans (Lm). causes blackleg disease on canola/rapeseed in many parts of the world. It is important to use resistant cultivars to manage the disease and minimize yield losses. In this study, twenty-two Lm isolates were used to identify resistance genes in a collection of 243 canola/rapeseed (Brassica napus L.) accessions from Canada and China. These Lm isolates carry different compliments of avirulence genes, and the investigation was based on a genome-wide association study (GWAS) and genotype-by-sequencing (GBS).

Results: Using the CROP-SNP pipeline, a total of 81,471 variants, including 78,632 SNPs and 2839 InDels, were identified. The GWAS was performed using TASSEL 5.0 with GLM + Q model. Thirty-two and 13 SNPs were identified from the Canadian and Chinese accessions, respectively, tightly associated with blackleg resistance with P values < 1 × 10^{- 4}. These SNP loci were distributed on chromosomes A03, A05, A08, A09, C01, C04, C05, and C07, with the majority of them on A08 followed by A09 and A03. The significant SNPs identified on A08 were all located in a 2010-kb region and associated with resistance to 12 of the 22 Lm isolates. Furthermore, 25 resistance gene analogues (RGAs) were identified in these regions, including two nucleotide binding site (NBS) domain proteins, fourteen RLKs, three RLPs and six TM-CCs. These RGAs can be the potential candidate genes for blackleg resistance.

Conclusion: This study provides insights into potentially new genomic regions for discovery of additional blackleg resistance genes. The identified regions associated with blackleg resistance in the germplasm collection may also contribute directly to the development of canola varieties with novel resistance genes against blackleg of canola.

Keywords: Blackleg; Brassica napus; Genome-wide association study (GWAS); Genotyping-by-sequencing (GBS); Leptosphaeria maculans; Phoma stem canker; Resistance gene analogues (RGAs).

BACKGROUND

Personal genome analysis is now being considered for evaluation of disease risk in healthy individuals, utilizing both rare and common variants. Multiple scores have been developed to predict the deleteriousness of amino acid substitutions, using information on the allele frequencies, level of evolutionary conservation, and averaged structural evidence. However, agreement among these scores is limited and they likely over-estimate the fraction of the genome that is deleterious.

METHODS

This study proposes an integrative approach to identify a subset of homozygous non-synonymous single nucleotide polymorphisms (nsSNPs). An 8-level classification scheme is constructed from the presence/absence of deleterious predictions combined with evidence of association with disease or complex traits. Detailed literature searches and structural validations are then performed for a subset of homozygous 826 mis-sense mutations in 575 proteins found in the genomes of 12 healthy adults.

RESULTS

Implementation of the Association-Adjusted Consensus Deleterious Scheme (AACDS) classifies 11% of all predicted highly deleterious homozygous variants as most likely to influence disease risk. The number of such variants per genome ranges from 0 to 8 with no significant difference between African and Caucasian Americans. Detailed analysis of mutations affecting the APOE, MTMR2, THSB1, CHIA, αMyHC, and AMY2A proteins shows how the protein structure is likely to be disrupted, even though the associated phenotypes have not been documented in the corresponding individuals.

CONCLUSIONS

The classification system for homozygous nsSNPs provides an opportunity to systematically rank nsSNPs based on suggestive evidence from annotations and sequence-based predictions. The ranking scheme, in-depth literature searches, and structural validations of highly prioritized mis-sense mutations compliment traditional sequence-based approaches and should have particular utility for the development of individualized health profiles. An online tool reporting the AACDS score for any variant is provided at the authors' website.