Profiles Of Secretions Research Articles

Comparative analysis of protein profiles in skin secretions of some Rana species: Preliminary insights into antimicrobial activity

Protein profiles of skin secretions of Rana dalmatina (Agile Frog), Rana macrocnemis (Uludağ Frog), Rana tavasensis (Tavas Frog) and Rana holtzi (Taurus Frog) frog species belonging to the Rana genus distributed in the Anatolian region of Türkiye were determined for the first time using the Tricine-SDS-PAGE Electrophoresis method and Coomassie Brilliant Blue (CBB) staining. By the results, some peptides with mass ≤5 kDa were detected. Just one peptide with mass ≤5 kDa was found in the secretion of each R. dalmatina, R. macrocnemis, and R. tavasensis while there was two in R. holtzi secretion. The antibacterial activity of secretions was determined using plate well diffusion assay on E. coli, S. typhimurium, S. aureus, B. cereus and L. monocytogenes bacteria. R. dalmatina created the inhibition zone for S. typhimurium, S. aureus, B. cereus, and L. monocytogenes. The zones of inhibition by R. tavasensis and R. macrocnemis species secretions were observed on S. aureus, B. cereus, and L. monocytogenes. It was found that R. holtzi creates an inhibition zone only on B. cereus. The results showed that the secretion of none of the species doesn't have antibacterial activity on E. coli. The skin secretion of R. dalmatina showed the most activity against bacteria, while R. holtzi had the least.

O-028 DNA methylation testing with cervical secretion can serve as a non-invasive biomarker for predicting successful implantation in frozen-thawed embryo transfer

Abstract Study question Is cervical secretion gene methylation profile associated with embryo implantation outcome, and can be used as a non-invasive biomarker for pregnancy prediction in FET cycles? Summary answer Cervical secretion gene methylation profiles show significant associations with implantation outcomes, and can serve as a non-invasive predictor of successful pregnancy in FET cycles. What is known already Cause of implantation failure remains a black box in reproductive medicine, and the exact mechanism of how endometrial receptivity is still behind the curtain. Epigenetic modifications including DNA methylation regulate gene expression and may alter endometrium receptivity. Endometrial DNA methylation changes vigorously only during the reconstruction of a new endometrium throughout each menstrual cycle, suggesting a unique methylation landscape in each regenerated endometrium. Cervical secretions contain implantation-related cytokines and genetic material from endometrium, and the DNA methylation profiles of cervical secretions is highly correlated with endometrium. Therefore, cervical secretions can be used as non-invasive surrogates for investigating endometrial condition. Study design, size, duration This multi-centered, case-control study enrolled 72 infertile females who entered the FET cycle with at least one high-quality blastocyst available between Aug 2017 and Dec 2022. Embryos assessed by Gardner scoring system with grade 3BB or above were defined as high-quality. Cervical secretions were collected on the day right before embryo transfer, and then DNA methylation profile were assessed. Participants/materials, setting, methods These females aged between 20-45 years-old with regular menstruation cycle (24-35 days) and normal BMI (17.5–32.0 kg/m2). Uterine cavity lesions and hydrosalpinx without treatment, adenomyosis, large uterine myoma that distorted endometrium, and thin endometrium (<7mm) were excluded. The association between the cervical secretion gene methylation profile and pregnancy outcome was evaluated via quantitative methylation-specific PCR (qMSP), methylation array, and genome-wide analysis. Different machine learning algorithms were utilized for differential methylation analysis and implantation prediction. Main results and the role of chance Cervical secretion DNA methylome differed between receptive (pregnant) and non-receptive (non-pregnant) FET cycle with genome-wide methylation array. The accuracy of pregnancy outcome prediction using top 2000 differential methylation probes (DMPs) in promoter region can be as high as 86% in all tested sample and 96.4% in pregnant samples. Bioinformatic analysis of six selected candidate genes related to implantation from literature review showed significant differences in DNA methylation between receptive and non-receptive endometrium. The methylation level of the six candidate genes with eight regions were measured by the qMSP platform and showed varying degrees of differential methylation status between the pregnancy and non-pregnancy groups. The AUCs for a single gene/region ranged from 0.53–0.67 for implantation prediction. Ten different machine learning models were applied and the results showed that the combination of candidate genes’ DNA methylation profiles could differentiate pregnant from non-pregnant samples with an accuracy as high as 86.67% and an AUC of 0.81 by logistic regression model. The best model from different machine learning algorithms using selected DMPs of genes related to window of implantation (WOI) can predict ongoing pregnancy with accuracy rates as high as 86.78% and AUC values up to 0.91. Limitations, reasons for caution The results obtained from a relatively small cohort size, and the timing of sample collection is on the day right before embryo transfer. Further investigation of methylation profiles in late follicular phase along with large sample size with euploid embryo transfer are mandatory for validation and clinical applications. Wider implications of the findings We firstly demonstrated the feasibility of cervical secretion methylation profile as a non-invasive testing for pregnancy prediction. The promising result may contribute to predicting endometrial receptivity for cycles with a favorable endometrium and maximize the successful pregnancy rate of embryo transfer. Trial registration number not applicable

Heat-altered scent marks of males of a fossorial reptile still allow recognition by females but lose information on male quality

The efficacy of sexual signals in communication is often maximized under specific environmental conditions. Anthropic alterations of these conditions might, thus, negatively affect communication during reproductive behavior. In fossorial animals, inhabiting visually restricted environments, chemical senses are very important. We examined whether climatic episodes of heat-waves with unusual high temperatures may affect the information provided to females by the sexual chemical signals of males of a fossorial reptile, the amphisbaenian Blanus cinereus. The results showed that experimentally heat-altered substrate scent marks of males can still provide information to females about the presence of a male. Females spent more time on males’ scent marks, irrespective of the temperature treatment, than on control clean ones. However, heat-altered scent marks did not seem to convey information about the health state (immune response) of the producer. Females spent more time on unaltered scent marks of healthier males (probably indicating mating preferences for these males), while female preferences for some heat-altered scent marks were not related to size or immune response of the same individual males. Chemical analyses indicated that the overall chemical profile of precloacal secretions (used for scent marking) did not change with increased temperatures. However, the relationship between proportions of some compounds in secretions and males’ immune response found in unaltered secretions was lost in heat-altered ones. We conclude that unusual increased environmental temperatures may decrease the efficacy of underground sexual chemical signals in this amphisbaenian (i.e., a loss of information on male quality), and consequently, may negatively affect sexual selection and reproduction.

Volatiles of symbiotic bacterial origin explain ectoparasitism and fledging success of hoopoes

BackgroundSome parasites use olfactory cues to detect their hosts and, since bacterial symbionts are partially responsible for animal odours, they could influence host parasitism. By autoclaving nest materials of hoopoe (Upupa epops) nests before reproduction started, we explored the hypothetical links between host-associated bacteria, volatiles and parasitism. During the nestling stage, we (i) estimated the level of ectoparasitism by chewing lice (Suborder Mallophaga) in adult hoopoe females and by Carnus haemapterus flies in nestlings, and (ii) characterized microbial communities and volatile profiles of nest environments (nest material and nest cavity, respectively) and uropygial secretions.ResultsExperimental nests had less diverse bacterial communities and more diverse volatile profiles than control nests, while occupants experienced lower intensity of parasitism in experimental than in control nests. The experiment also affected beta diversity of the microbial communities of nest material and of the volatiles of the nestling uropygial secretions. Moreover, microbial communities of uropygial secretions and of nest materials covaried with their volatile profiles, while the volatile profile of the bird secretions explained nest volatile profile. Finally, a subset of the volatiles and bacteria detected in the nest material and uropygial secretions were associated with the ectoparasitism intensity of both adult females and nestlings, and with fledging success.ConclusionsThese results show that a component of animal odours is linked with the microbial communities of the host and its reproductive environment, and emphasize that the associations between bacteria, ectoparasitism and reproductive success are partially mediated by volatiles of bacterial origin. Future work should focus on mechanisms underlying the detected patterns.

Cutaneous glands of the striped toad, Rhinella crucifer (Wied-Neuwied, 1821) (Amphibia: Bufonidae): Histological study and bioactivities of glandular secretions

This study investigated the morphology of Rhinella crucifer cutaneous glands, as well as the protein/peptide profiles and bioactivities of body gland secretions (BGS) and parotoid macrogland secretions (PS). The parotoid as well as dorsal and ventral skin fragments of male and female individuals were processed for histological analysis. The protein and peptide profiles of male and female gland secretions were evaluated. Male secretions were also assessed for proteolytic, trypsin inhibiting, hemagglutinating, hemolytic, antimicrobial, and anticoagulant activities. The R. crucifer skin structure presented protuberances that are clearly visible and formed by the integument, which has cutaneous glands throughout the body. An average of 438 and 333 glands were identified in males in females, respectively. No significant differences were observed in the distribution of glands across the body as well as for area and perimeter of glands. Differences were observed in protein composition between the PS and BGS from males and females, and secretions from animals collected from undisturbed and anthropogenically disturbed areas. Proteins with similarities to catalase and elongation factor 1-alpha were detected in the PS. Zymography revealed proteolytic activity in both male BGS and PS. Male BGS showed antibacterial activity against Enterococcus faecalis and Escherichia coli and anticoagulant activity, being able to prolong prothrombin time by 6.34-fold and activated partial thromboplastin time by 2.17-fold. Finally, male PS and BGS caused a maximum hemolysis degree of 1.4%. The data showed that the cutaneous secretions of R. crucifer are potentially promising for biotechnological prospecting.

Dynamics of the content of immunoglobulin classes in blood serum and vaginal secretions of cows with genital mycoplasmosis with various methods of therapy

The economic efficiency of industrial animal husbandry in modern conditions provides not only for an increase in productivity, but also for maintaining the optimal level of basic physiological functions, including reproductive ones. The successful implementation of the task of preserving reproductive function is hindered by diseases of females of various etiologies that negatively affect their reproductive ability. One of these diseases is genital mycoplasmosis of cattle, which has a significant prevalence on dairy farms, both abroad and in the Russian Federation. The effectiveness of therapy for this disease largely depends on the initial state of the immune system. The objective of our research was to study the therapeutic efficacy and effect on the immunoglobulin composition of blood serum and vaginal secretions of patients with genital mycoplasmosis of cows of the use of the antibiotic tulathromycin and its combination with the immunomodulator thymalin. The results obtained demonstrated that the use of tulatromycin for treatment leads to the recovery of 75% of experimental animals and ensures the restoration of the immunoglobulin profile of vaginal secretions, whereas the immunodeficiency state observed in blood serum is only partially restored. The combination of an antibiotic and an immunomodulator ensured the recovery of all experimental animals and the restoration of the immunological characteristics of blood serum and vaginal secretions to the level of healthy cows.

Comprehensive Analysis of Bufadienolide and Protein Profiles of Gland Secretions from Medicinal Bufo Species.

Toad Venom (TV) is the dried product of toxic secretions from Bufo bufo gargarizans Cantor (BgC) or B. melanostictus Schneider (BmS). Given the increasing medical demand and the severe depletion of wild toads, a number of counterfeit TVs appeared on the market, posing challenges to its quality control. In order to develop an efficient, feasible, and comprehensive approach to evaluate TV quality, a thorough analysis and comparison of chemical compounds among legal species BgC and BmS, as well as the main confusion species B. andrewsi Schmidt (BaS) and B. raddei Strauch (BrS), were conducted by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), high performance liquid chromatography (HPLC), sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Nano LC-MS/MS analyses. We identified 126 compounds, including free or conjugated bufadienolides, indole alkaloids and amino acids, among the four Bufo species. The content of main bufadienolides, such as gamabufotalin, bufotalin, bufalin, cinobufagin, and resibufogenin, and the total protein contents varied widely among 28 batches of TV due to their origin species. The sum of the five bufadienolides within the BgC, BmS, BaS, and BrS samples were 8.15-15.93%, 2.45-4.14%, 11.15-13.50%, and 13.21-14.68%, respectively. The total protein content of BgC (6.9-24.4%) and BaS (19.1-20.6%) samples were higher than that of BmS (4.8-20.4%) and BrS (10.1-13.7%) samples. Additionally, a total of 1357 proteins were identified. There were differences between the protein compositions among the samples of the four Bufo species. The results indicated that BgC TV is of the highest quality; BaS and BrS TV could serve as alternative resources, whereas BmS TV performed poorly overall. This research provides evidence for developing approaches to evaluate TV quality and selecting the proper Bufo species as the origin source of TV listed in the Chinese pharmacopoeia.

Sex differences in peripheral immune cell activation: Implications for pain and pain resolution

Decades of research into chronic pain has deepened our understanding of the cellular mechanisms behind this process. However, a failure to consider the biological variable of sex has limited the application of these breakthroughs into clinical application. In the present study, we investigate fundamental differences in chronic pain between male and female mice resulting from inflammatory activation of the innate immune system. We provide evidence that female mice are more sensitive to the effects of macrophages. Injecting small volumes of media conditioned by either unstimulated macrophages or macrophages stimulated by the inflammatory molecule TNFα lead to increased pain sensitivity only in females. Interestingly, we find that TNFα conditioned media leads to a more rapid resolution of mechanical hypersensitivity and altered immune cell recruitment to sites of injury. Furthermore, male and female macrophages exhibit differential polarization characteristics and motility after TNFα stimulation, as well as a different profile of cytokine secretions. Finally, we find that the X-linked gene Tlr7 is critical in the facilitating the adaptive resolution of pain in models of acute and chronic inflammation in both sexes. Altogether, these findings suggest that although the cellular mechanisms of pain resolution may differ between the sexes, the study of these differences may yield more targeted approaches with clinical applications.

Effects of pharmacological castration on endocrinological and chemical profiles in captive red and fallow deer

In wildlife captive management, contraception is highly desirable due to inbreeding risk and needed control of intra- and inter-specific aggressions. Deer, for which surgical castration can cause abnormal growth of the antlers, are suitable candidates for pharmacological castration. The aim of this study was to evaluate the effect of pharmacological castration on the physiological patterns and volatile chemical profiles of two deer species. We placed a subcutaneous implant of deslorelin acetate (4.7 mg) on four male red deer (Cervus elaphus) and two male fallow deer (Dama dama) (N = 6) living together in a mixed-sex and -species group housed at a wildlife rescue centre in Tuscany, Italy. We combined hormone measurements and chemical investigation of metatarsal scent-gland odour secretions to test whether pharmacological castration will influence the volatile chemical profile, potentially modifying the olfactory communication during the rutting season. We used radioimmunoassay technique to determine faecal and plasma testosterone as well as faecal cortisol levels, and solid-phase microextraction coupled to gas chromatography-mass spectrometry to analyse the scent-gland odour secretions. We found that deer testosterone levels decreased after the subcutaneous deposition of the implant. However, castrated red deer exhibited more variability in testosterone concentrations throughout the study period compared to fallow deer, whose testosterone levels remained low after the implant. We detected a total of 124 (red deer) and 88 (fallow deer) volatile compounds in male scent-gland odour secretions, including naturally occurring ketones, alcohols, aldehydes, terpenes, carboxylic acids, esters, volatile fatty acids, and hydrocarbons. Odour richness, but not total abundance or ratios of compounds, changed with testosterone levels in red deer. In conclusion, our preliminary findings suggest that deslorelin acetate, causing a decrease of testosterone level and impacting the chemical profile of odour secretions, could work as a non-invasive contraception approach in human-managed populations of deer.

Brown bear skin-borne secretions display evidence of individuality and age-sex variation

Scent originates from excretions and secretions, and its chemical complexity in mammals translates into a diverse mode of signalling. Identifying how information is encoded can help to establish the mechanisms of olfactory communication and the use of odours as chemical signals. Building upon existing behavioural and histological literature, we examined the chemical profile of secretions used for scent marking by a solitary, non-territorial carnivore, the brown bear (Ursus arctos). We investigated the incidence, abundance, and uniqueness of volatile organic compounds (VOCs) from cutaneous glandular secretions of 12 wild brown bears collected during late and post-breeding season, and assessed whether age-sex class, body site, and individual identity explained profile variation. VOC profiles varied in the average number of compounds, compound incidence, and compound abundance by age-sex class and individual identity (when individuals were grouped by sex), but not by body site. Mature males differed from other age-sex classes, secreting fewer compounds on average with the least variance between individuals. Compound uniqueness varied by body site and age for both males and females and across individuals. Our results indicate that brown bear skin-borne secretions may facilitate age-sex class and individual recognition, which can contribute towards further understanding of mating systems and social behaviour.

Comparison of volatile compounds of anal sac secretions between the sexes of domestic dog (Canis lupus familiaris).

Volatile compounds of anal sac secretions are odorant chemicals used across the carnivores for social communication such as identifying individuals and group membership. Odor profiles taken from expressed anal sac secretions of some species of carnivores have been detected in previous studies. In this study, the volatile compounds of anal sac secretions between five male and five female domestic dogs (Canis lupus familiaris) were compared. Volatile chemicals were extracted, separated, and analyzed by gas chromatography-mass spectrometry with solid-phase micro-extraction and identified from their electron ionization mass spectra and Kovats retention indices. The results showed the presence of various types of compounds including organic fatty acids, ketones, aldehydes, esters, and alcohols in the anal sac secretions of dogs. Greater amounts of diversity and esters, and lower amounts of alcohols were detected in the anal sac secretions of females compared to males. This was accompanied by finding citrate and acetic acid ester only in the females. Furthermore, presence of some sex-specific organic compounds like dimethylcyclopentyl ethanone indicates that the volatile profiles of anal sac secretions in 10 domestic dogs are differentiated by host sex.

Bacteriological Profile and Antibiotic Susceptibility Pattern of Tracheal Secretions Isolates Among Intensive Care Unit Patients at Tertiary Care Hospital

Introduction: Respiratory infections are associated with high morbidity and mortality, especially in critically ill patients. The excessive use of broad-spectrum antibiotics has led to the development of drug resistance, thus resulting in the emergence of pathogens which are difficult to treat. Materials and Methods: A total of 632 tracheal secretions were received in the Bacteriology section of the microbiology department of a tertiary care hospital from November 2019 to February 2020. Tracheal secretions were processed for culture according to standard operating procedures. Identification, phenotype detection, and antibiotic sensitivity testing were performed by automated VITEK-2 Compact system. Results: In total, 632 tracheal secretions were received during the study period, 559 cultures yielded significant pathogens and no organisms were isolated in 73 cultures. Among the Gram-negative organism 540 (97%), Klebsiella pneumoniae (30%) was the most common isolates. Gram-positive organisms 12 (2%) and Candida spp. 7 (1%) were isolated. The most common phenotype detected in Escherichia coli and K. pneumoniae was extended-spectrum beta-lactamase producer. Conclusions: K. pneumoniae was the most common isolate from tracheal secretion among intensive care unit patients. Colistin, followed by tigecycline, was found to be the most susceptible antibiotics. K. pneumoniae was found to be sensitive to tigecycline (69%) with minimum inhibitory concentrations of ≤ 1. 0.6%. K. pneumoniae was colistin resistant.

Anogenital scent-marking signals fertility in a captive female Alaotran gentle lemur

The Lake Alaotra gentle lemur (Hapalemur alaotrensis) is one of the 25 most endangered primates in the world and shows low success rate in captive breeding programmes. It is therefore vital to further understand its reproductive biology. We studied a captive troop consisting of five individuals hosted at Jersey Zoo during breeding and non-breeding periods over 1 year. We collected behavioural data (n = 318 h) using all occurrence of some behaviours and ad libitum sampling methods, as well as faecal (n = 54) and anogenital scent (n = 35) samples of the breeding female. We measured sex hormone levels using enzyme immunoassay technique and investigated the volatile component of odour signals using solid-phase microextraction and gas chromatography-mass spectrometry. We observed sexual and aggressive behaviours occasionally during the breeding period. Our regression analysis showed that only period significantly predicted rates of female anogenital scent-marking, whereby the female performed anogenital scent-marking more frequently during the breeding rather than the non-breeding period. In contrast, female hormone levels did not significantly explain variation in rates of neither male nor female olfactory, sexual and affiliative behaviours, suggesting that individuals' behaviour alone is not an effective indicator of the ovulation window. The volatile chemical profile of anogenital odour secretions changed over the study, with four compounds distinguishing the fertile window during the breeding period. In conclusion, our findings suggest that anogenital scent-marking may signal the reproductive status of captive female gentle lemurs.

Genome-wide analysis of cervical secretions obtained during embryo transfer reveals the association between deoxyribonucleic acid methylation and pregnancy outcomes

To study whether the methylation status of cervical secretions can reflect the ability of the endometrium to allow embryo implantation. Case-control study. Invitro fertilization centers. Women undergoing embryo transfer cycles, in which at least 1 good-quality embryo was transferred. Collection of cervical secretions during the procedure of embryo transfer. Methylation profiles of cervical secretions in relation to pregnancy outcomes. Genome-wide methylation profiles differ between cervical secretions from pregnancy and nonpregnancy cycles. Clustering analysis on the basis of the top 2,000 differentially methylated probes of cervical secretions from 28 pregnancy and 29 nonpregnancy cycles correctly categorized 86.0% of the samples in terms of conceptional status, which was verified in selected genes by quantitative methylation-specific polymerase chain reaction and validated in another independent sample set. The combination of selected genes was estimated to predict pregnancy outcomes with a maximal area under the receiver operating characteristic curve of 0.83. The methylation profiles of cervical secretions were associated with pregnancy outcomes in embryo transfer cycles. Although not clinically useful at present, deoxyribonucleic acid methylation in cervical secretions may shed new light on the less invasive assessment of endometrial receptivity.

Altered immunometabolism in adipose tissue: A major contributor to the ageing process?

Altered immunometabolism in adipose tissue: A major contributor to the ageing process?

Microbiological profile and sensitivity pattern of endotracheal secretions in mechanically ventilated patients

Endotracheal intubation and mechanical ventilation are life-saving procedures done on emergency or elective basis to prevent or combat respiratory failure. Many clinical conditions warrant need for ventilatory support like, life threatening infections, sepsis and acute respiratory distress syndromes, neurological dysfunctions due to poisoning, drug toxicity, cerebrovascular accidents, trauma and others. The presence of an endotracheal tube in the airway, although critical for the management of the mechanically ventilated patient, also contributes to the development of ventilator associated pneumonia by disrupting normal protective mechanism which is associated with the intraluminal formation of biofilm by multidrug resistant organisms. Hence in our study endotracheal secretions were sent for bacteriological culture and sensitivity on the first day of intubation to identify the organisms that already existed at the time of intubation which would help in initiating and or modifying antibiotic therapy appropriately.

Abstract 13545: Atlas of Exosomal Microrna Secreted From Human Ipsc-derived Cardiac Cell Types

Exosomes are small particles secreted from cells through an endosomal sorting complexes required for transport (ESCRT)-mediated pathway that releases vesicles into the extracellular matrix following fusion of the endosomal multivesicular body to the plasma membrane. Initially thought to be a waste disposal system for the cell, exosomes are now recognized as important mediators in cellular communication, transporting nucleic acid, protein and lipid. We hypothesize that microRNA (miRNA) transcriptomic profiles isolated from exosomes are distinct for each cardiovascular lineage in healthy control patients. Using induced pluripotent stem cells (iPSC), established differentiation protocols and a high-yield mechanical-sorting device (ExoTIC), we characterized the transcriptomic profiles of exosomal secretions derived from cardiomyocytes (CM), cardiac fibroblasts (CF) and endothelial cells (EC). We identified 120 miRNAs from 94 miRNA genes to be secreted at appreciable levels. We discovered 1) let-7 miRNA precursors are depleted in iPSC secretomes; 2) only a subset of total cellular miRNAs are present; 3) a common core of miRNAs are secreted by all three cardiac cell types; and 4) distinct enrichment or depletion of different mature miRNAs in each exosome type. The exosomal transcriptomic profiles of each lineage was compared to a published atlas of miRNA from seventeen tissue types from the human body. miR-302c known to be specific to pluripotent cells and is enriched in iPSC exosomes and found in bone, reflecting hematopoietic stem cells. miR-1, critical for cardiac development and pathology, is secreted by iPSC-CM and enriched in striated muscle and myocardium. miR-151 is secreted by iPSC-CF and is expressed ubiquitously. The relative enrichment of miRNAs in different cardiac tissues could be used to quantifying the cellular content of heterogenous mixtures of iPSC-derived cardiac lineages. We developed a ratiometric cell purity assay that can sensitively detect iPSC contamination in iPSC-CM differentiations.

Proteome analysis of secretions from human monocyte-derived macrophages post-exposure to biomaterials and the effect of secretions on cardiac fibroblast fibrotic character

The use of exogenous biomolecules (BM) for the purpose of repairing and regenerating damaged cardiac tissue can yield serious side effects if used for prolonged periods. As well, such strategies can be cost prohibitive depending on the regiment and period of time applied. Alternatively, autologous monocytes/monocyte-derived macrophages (MDM) can provide a viable path towards generating an endogenous source of stimulatory BM. Biomaterials are often considered as delivery vehicles to generate unique profiles of such BM in tissues or to deliver autologous cells, that can influence the nature of BM produced by the cells. MDM cultured on a degradable polar hydrophobic ionic (D-PHI) polyurethane has previously demonstrated a propensity to increase select anti-inflammatory cytokines, and therefore there is good rationale to further investigate a broader spectrum of the cells’ BM in order to provide a more complete proteomic analysis of human MDM secretions induced by D-PHI. Further, it is of interest to assess the potential of such BM to influence cells involved in the reparative state of vital tissues such as those that affect cardiac cell function. Hence, this current study examines the proteomic profile of MDM secretions using mass spectrometry for the first time, along with ELISA, following their culture on D-PHI, and compares them to two important reference materials, poly(lactic-co-glycolic acid) (PLGA) and tissue culture polystyrene (TCPS). Secretions collected from D-PHI cultured MDM led to higher levels of regenerative BM, AGRN, TGFBI and ANXA5, but lower levels of pro-fibrotic BM, MMP7, IL-1β, IL-6 and TNFα, when compared to MDM secretions collected from PLGA and TCPS. In the application to cardiac cell function, the secretion collected from D-PHI cultured MDM led to more human cardiac fibroblast (HCFs) migration. A lower collagen gel contraction induced by MDM secretions collected from D-PHI was supported by gene array analysis for human fibrosis-related genes. The implication of these findings is that more tailored biomaterials such as D-PHI, may lead to a lower pro-inflammatory phenotype of macrophages when used in cardiac tissue constructs, thereby enabling the development of vehicles for the delivery of interventional therapies, or be applied as coatings for sensor implants in cardiac tissue that minimize fibrosis. The general approach of using synthetic biomaterials in order to induce MDM secretions in a manner that will guide favorable regeneration will be critical in making the choice of biomaterials for tissue regeneration work in the future. Statement of significanceImmune modulation strategies currently applied in cardiac tissue repair are mainly based on the delivery of defined exogenous biomolecules. However, the use of such biomolecules may pose wide ranging systemic effects, thereby rendering them clinically less practical. The chemistry of biomaterials (used as a potential targeted delivery modality to circumvent the broad systemic effects of biomolecules) can not only affect acute and chronic toxicity but also alters the timeframe of the wound healing cascade. In this context, monocytes/monocyte-derive macrophages (MDM) can be harnessed as an immune modulating strategy to promote wound healing by an appropriate choice of the biomaterial. However, there are limited reports on the complete proteome analysis of MDM and their reaction of biomaterial related interventions on cardiac tissues and cells. No studies to date have demonstrated the complete proteome of MDM secretions when these cells were cultured on a non-traditional immune modulatory ionomeric polyurethane D-PHI film. This study demonstrated that MDM cultured on D-PHI expressed significantly higher levels of AGRN, TGFBI and ANXA5 but lower levels of MMP7, IL-1β, IL-6 and TNFα when compared to MDM cultured on a well-established degradable biomaterials in the medical field, e.g. PLGA and TCPS, which are often used as the relative standards for cell culture work in the biomaterials field. The implications of these findings have relevance to the repair of cardiac tissues. In another aspect of the work, human cardiac fibroblasts showed significantly lower contractility (low collagen gel contraction and low levels of ACTA2) when cultured in the presence of MDM secretions collected after culturing them on D-PHI compared to PLGA and TCPS. The findings place emphasis on the importance of making the choice of biomaterials for tissue engineering and regenerative medicine applied to their use in cardiac tissue repair.

Comparative study of the chemical profile of the parotoid gland secretions from Rhaebo guttatus from different regions of the Brazilian Amazon

Amphibian cutaneous secretion has great potential for bioprospection and is a great tool in the development of bioproducts. Thus, the objective of the present work was to evaluate the comparative study of the chemical profile parotoid gland secretions from Rhaebo guttatus collected in two distinct regions of the Brazilian Amazon. For this, the chemical composition of six methanolic extracts of this species were analyzed by Liquid Chromatography in UV and MS Detection Ultra-Chromatography Systems (UFLC-DAD-micrOTOF). All obtained chromatograms presented two distinct regions; one referring to the more hydrophilic molecules (alkaloids), while the other refers to the more hydrophobic compounds (steroids). The steroid region resembles all samples, regardless of where they were collected. In the alkaloid region, there was a standardized variation for the samples from the southern Brazilian Amazon, but the same was not true for the samples collected in the Amazon-Cerrado transition region. Thus, the data suggest that the environment and diet of R. guttatus may be important in alkaloid production, but do not influence steroid content. These results add new information about the poison of the toad R. guttatus and raises new questions to be further investigated, thus contributing to the knowledge of the anuran fauna of the Brazilian Amazon.

MicroRNA profiles of dry secretions through the first three weeks of the dry period from Holstein cows

In dairy cows, the period from the end of lactation through the dry period and into the transition period, requires vast physiological and immunological changes critical to mammary health. The dry period is important to the success of the next lactation and intramammary infections during the dry period will adversely alter mammary function, health and milk production for the subsequent lactation. MicroRNAs (miRNAs) are small non-coding RNAs that can post transcriptionally regulate gene expression. We sought to characterize the miRNA profile in dry secretions from the last day of lactation to 3, 10, and 21 days post dry-off. We identified 816 known and 80 novel miRNAs. We found 46 miRNAs whose expression significantly changed (q-value < 0.05) over the first three weeks of dry-off. Additionally, we examined the slopes of random regression models of log transformed normalized counts and cross analyzed the 46 significantly upregulated and downregulated miRNAs. These miRNAs were found to be associated with important components of pregnancy, lactation, as well as inflammation and disease. Detailing the miRNA profile of dry secretions through the dry-off period provides insight into the biology at work, possible means of regulation, components of resistance and/or susceptibility, and outlets for targeted therapy development.