Methyl and isopropyl N-methylanthranilates attenuate diclofenac- and ethanol-induced gastric lesions in rats

Aims

Two natural alkaloids, methyl (M) and isopropyl (I) N-methylanthranilates, with recently demonstrated significant pharmacological activities, were assayed for their possible overall effect on intact gastric mucosa and their protective properties towards the onset of gastric lesions induced by diclofenac (a non-steroidal anti-inflammatory drug, NSAID) or ethanol.

Main methods

The influence of I and M on gastric mucosa integrity was assessed by oral administration in doses of 200 mg/kg. The gastroprotective action of I and M in doses of 50, 100 and 200 mg/kg was analyzed in the diclofenac and ethanol-induced gastric lesion models in rats. After the treatment, the stomachs of the animals were analyzed (captured by a digital camera). Ulcer scoring, morphometric and histopathological analyses of the stomachs were done.

Key findings

The oral application of these compounds on their own, even in quite high doses (200 mg/kg) did not induce gastric lesions. Both alkaloids exerted a very strong antiulcer activity, even in low doses (50 mg/kg), by decreasing the number of lesions caused by the application of either diclofenac or ethanol, eliminating them completely or reducing them to a form of mucosal hyperemia.

Significance

Their possible mechanism of action was discussed and due to their many positive properties including anxiolytic, antidepressant, antinociceptive, anti-inflammatory and gastroprotective activities, as well as a cheap and simple synthetic route for their preparation, methyl and isopropyl N-methylanthranilates, both alike, might represent a cost effective alternative sought for in the treatment of peptic ulcers and/or new safer NSAIDs for pain management.     

The Incidence of Human Cysticercosis in a Rural Community of Eastern Zambia

A community-based longitudinal study was performed in the Eastern Province of Zambia, in which repeated serological samplings were done to determine the incidence of human cysticercosis. Three sampling rounds were carried out at six months intervals. A total of 867 participants presented for all three samplings. All samples were tested for the presence of cysticercus antigens using a monoclonal antibody-based enzyme-linked immunosorbent assay (sero-Ag-ELISA), while a randomly selected sub-sample of 161 samples from each sampling round was tested for specific antibodies using a commercial enzyme-linked immunoelectrotransfer blot (EITB) assay. Stool samples (n = 226) were also collected during the final round of sampling for taeniosis diagnosis by coprology and coproantigen ELISA. Cysticercosis seroprevalence varied from 12.2% to 14.5% (sero-Ag) and from 33.5% to 38.5% (sero-Ab) during the study period. A taeniosis prevalence of 11.9% was determined. Incidence rates of 6300 (sero-Ag, per 100000 persons-year) and 23600 (sero-Ab, per 100000 persons-year) were determined. Seroreversion rates of 44% for sero-Ag and 38.7% for sero-Ab were recorded over the whole period. In conclusion, this study has shown the dynamic nature of T. solium infections; many of the people at risk become (re)infected due to the high environmental contamination, with a high number turning seronegative within a year after infection. An important number of infections probably never fully establish, leading to transient antibody responses and short-term antigen presence.     

scAmpi—A versatile pipeline for single-cell RNA-seq analysis from basics to clinics

Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful technique to decipher tissue composition at the single-cell level and to inform on disease mechanisms, tumor heterogeneity, and the state of the immune microenvironment. Although multiple methods for the computational analysis of scRNA-seq data exist, their application in a clinical setting demands standardized and reproducible workflows, targeted to extract, condense, and display the clinically relevant information. To this end, we designed scAmpi (Single Cell Analysis mRNA pipeline), a workflow that facilitates scRNA-seq analysis from raw read processing to informing on sample composition, clinically relevant gene and pathway alterations, and in silico identification of personalized candidate drug treatments. We demonstrate the value of this workflow for clinical decision making in a molecular tumor board as part of a clinical study.     

World Health Organization Estimates of the Relative Contributions of Food to the Burden of Disease Due to Selected Foodborne Hazards: A Structured Expert Elicitation

Background

The Foodborne Disease Burden Epidemiology Reference Group (FERG) was established in 2007 by the World Health Organization (WHO) to estimate the global burden of foodborne diseases (FBDs). This estimation is complicated because most of the hazards causing FBD are not transmitted solely by food; most have several potential exposure routes consisting of transmission from animals, by humans, and via environmental routes including water. This paper describes an expert elicitation study conducted by the FERG Source Attribution Task Force to estimate the relative contribution of food to the global burden of diseases commonly transmitted through the consumption of food.

Methods and Findings

We applied structured expert judgment using Cooke’s Classical Model to obtain estimates for 14 subregions for the relative contributions of different transmission pathways for eleven diarrheal diseases, seven other infectious diseases and one chemical (lead). Experts were identified through international networks followed by social network sampling. Final selection of experts was based on their experience including international working experience. Enrolled experts were scored on their ability to judge uncertainty accurately and informatively using a series of subject-matter specific ‘seed’ questions whose answers are unknown to the experts at the time they are interviewed. Trained facilitators elicited the 5^th, and 50^th and 95^th percentile responses to seed questions through telephone interviews. Cooke’s Classical Model uses responses to the seed questions to weigh and aggregate expert responses. After this interview, the experts were asked to provide 5^th, 50^th, and 95^th percentile estimates for the ‘target’ questions regarding disease transmission routes. A total of 72 experts were enrolled in the study. Ten panels were global, meaning that the experts should provide estimates for all 14 subregions, whereas the nine panels were subregional, with experts providing estimates for one or more subregions, depending on their experience in the region. The size of the 19 hazard-specific panels ranged from 6 to 15 persons with several experts serving on more than one panel. Pathogens with animal reservoirs (e.g. non-typhoidal Salmonella spp. and Toxoplasma gondii) were in general assessed by the experts to have a higher proportion of illnesses attributable to food than pathogens with mainly a human reservoir, where human-to-human transmission (e.g. Shigella spp. and Norovirus) or waterborne transmission (e.g. Salmonella Typhi and Vibrio cholerae) were judged to dominate. For many pathogens, the foodborne route was assessed relatively more important in developed subregions than in developing subregions. The main exposure routes for lead varied across subregions, with the foodborne route being assessed most important only in two subregions of the European region.

Conclusions

For the first time, we present worldwide estimates of the proportion of specific diseases attributable to food and other major transmission routes. These findings are essential for global burden of FBD estimates. While gaps exist, we believe the estimates presented here are the best current source of guidance to support decision makers when allocating resources for control and intervention, and for future research initiatives.     

The association between flavor labeling and flavor recall ability in children

This study sought to determine if the ability to label a flavor is associated with an improved ability to recall having tasted the flavor in preschool-aged children. A total of 120 3- to 6-year-old English-speaking children tasted and labeled 20 different flavors, blinded to color. Children's labels for the flavors were scored for consistency and accuracy. Recall for having tasted the flavor was tested. Both labeling ability and recall ability improved rapidly between the ages of 3 and 6 years in this cohort. Regression analysis indicated that independent of the child's age, consistent accurate labeling was positively associated with recall ability. Higher maternal education was an independent and marginal contributor to greater recall ability. The combination of consistent and accurate labeling, age, and maternal education accounted for 28% of the variance in flavor recall ability. Consistent but inaccurate labeling alone contributed little to the variance in flavor recall ability. We conclude from these findings that children's ability to recall having tasted a flavor develops rapidly during the preschool age range and that improved recall ability is associated with the ability to consistently and accurately label the flavor. We conclude that language mediates memory for flavors in young children.     

Phospholipid transfer protein is present in human tear fluid

The human tear fluid film consists of a superficial lipid layer, an aqueous middle layer, and a hydrated mucin layer located next to the corneal epithelium. The superficial lipid layer protects the eye from drying and is composed of polar and neutral lipids provided by the meibomian glands. Excess accumulation of lipids in the tear film may lead to drying of the corneal epithelium. In the circulation, phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) mediate lipid transfers. To gain insight into the formation of tear film, we investigated whether PLTP and CETP are present in human tear fluid. Tear fluid samples were collected with microcapillaries. The presence of PLTP and CETP was studied in tear fluid by Western blotting, and the PLTP concentration was determined by ELISA. The activities of the enzymes were determined by specific lipid transfer assays. Size-exclusion and heparin-affinity chromatography assessed the molecular form of PLTP. PLTP is present in tear fluid, whereas CETP is not. Quantitative assessment of PLTP by ELISA indicated that the PLTP concentration in tear fluid, 10.9 +/- 2.4 microg/mL, is about 2-fold higher than that in human plasma. PLTP-facilitated phospholipid transfer activity in tears, 15.1 +/- 1.8 micromol mL(-)(1) h(-)(1), was also significantly higher than that measured in plasma. Inactivation of PLTP by heat treatment (+58 degrees C, 60 min) or immunoinhibition abolished the phospholipid transfer activity in tear fluid. Size-exclusion chromatography of tear fluid indicated that PLTP eluted in a position corresponding to a size of 160-170 kDa. Tear fluid PLTP was quantitatively bound to Heparin-Sepharose and could be eluted as a single peak by 0.5 M NaCl. These data indicate that human tear fluid contains catalytically active PLTP protein, which resembles the active form of PLTP present in plasma. The results suggest that PLTP may play a role in the formation of the tear film by supporting phospholipid transfer.     

Quantification of 1H NMR spectra of human cerebrospinal fluid: a protocol based on constrained total-line-shape analysis

A protocol for quantitative ¹H NMR analysis of human cerebrospinal fluid (hCSF) was built up and assessed as based on Constrained Total-Line-Shape (CTLS) fitting. In this method, linear constraints were applied to spectral structures. The ¹H NMR spectra of 45 human CSF samples were measured and quantified using the CTLS method. The quantification strategies based on total-line-shape fitting are discussed. The metabolic model for CTLS includes 31 metabolites covering 85% of the total spectral intensity, excluding the protein contribution. Prior to data analysis, the data was divided into patients with no Alzheimer’s disease (AD), but with a normal AD marker profile (the peptide β-amyloid₄₂ and tau protein) present in CSF, and into controls that do not have an AD marker profile in CSF. Unexpectedly large variations in metabolite concentrations within the two patient groups were detected, but an analysis of variance revealed a significant (P = 0.027) difference only in the concentration of creatinine which was higher in patients that had a normal AD marker profile. Multivariate classification tools such as self-organizing maps (SOM) failed in separation of the two classes.     

A plea for simultaneously considering matrix quality and local environmental conditions when analysing landscape impacts on effective dispersal

Landscape genetics has tremendous potential for enhancing our understanding about landscape effects on effective dispersal and resulting genetic structures. However, the vast majority of landscape genetic studies focus on effects of the landscape among sampling locations on dispersal (i.e. matrix quality), while effects of local environmental conditions are rather neglected. Such local environmental conditions include patch size, habitat type or resource availability and are commonly used in (meta‐) population ecology and population genetics. In our opinion, landscape genetic studies would greatly benefit from simultaneously incorporating both matrix quality and local environmental conditions when assessing landscape effects on effective dispersal. To illustrate this point, we first outline the various ways in which environmental heterogeneity can influence different stages of the dispersal process. We then propose a three‐step approach for assessing local and matrix effects on effective dispersal and review how both types of effects can be considered in landscape genetic analyses. Using simulated data, we show that it is possible to correctly disentangle the relative importance of matrix quality vs. local environmental conditions for effective dispersal. We argue that differentiating local and matrix effects in such a way is crucial for predicting future species distribution and persistence, and for optimal conservation decisions that are based on landscape genetics. In sum, we think it is timely to move beyond purely statistical, pattern‐oriented analyses in landscape genetics and towards process‐oriented approaches that consider the full range of possible landscape effects on dispersal behaviour and resulting gene flow.