A Comparative Study of Effects of Biodegradable and Non-biodegradable Microplastics on the Growth and Development of Black Soldier Fly Larvae (Hermetia illucens).

Purpose: This study aimed to investigate the digestion process of biodegradable and non-biodegradable microplastics (MPs) within black soldier fly larvae (BSFL) and assess their impact on larval growth and development. The goal was to understand the fate of MPs within BSFL, considering their potential for waste conversion polluted with MPs. Methods: BSFL were exposed to two types of MPs, and their growth, development, potential accumulation and excretion of MPs were monitored. Results: The findings revealed that the MPs accumulated solely in the larval gut and had no adverse effects on the growth and development of BSFL. Larvae efficiently excreted MPs before reaching the pupation stage. Conclusion: This research emphasizes the potential of BSFL as a bioconversion agent for organic waste, even in the presence of MPs. The effective excretion of MPs by BSFL before pupation suggests their ability to mitigate potential harm caused by MP accumulation. The fact that BSFL may excrete MPs before pupation would contribute to their safe use as animal feedstock. A careful evaluation of the effects of using BSFL reared on contaminated substrates especially containing visually non-detectable residuals like nanoplastics, chemicals or toxic metals and further examination of the broader implications for waste management and sustainable livestock farming remains important. Graphical Abstract: Experimental design outlining the workflow for the analyses used to investigate the effect of two types of microplastics, polyamide (PA), and polylactic acid (PLA), on growth and development of black soldier fly larvae.

Performance of a predictive weight loss model in terms of rapid detection of inadequate weight loss after Roux-en-Y gastric bypass.

BACKGROUND: Predictive weight loss models can help patients meet their expectations after bariatric surgery and assist physicians in responding to deviations from the predicted weight. A model published by Seyssel et al. appears to accurately predict postoperative body mass index. OBJECTIVES: We aimed to demonstrate this model's performance in terms of rapid detection of insufficient weight loss (surgical nonresponse). SETTING: Cantonal Hospital, Switzerland. METHODS: We retrospectively analyzed weight and body mass index deviations at 2 years postoperatively, based on values predicted by the model of Seyssel et al. The primary outcome was the timing of detection of surgical nonresponse. The secondary outcome was how patients' weight loss expectations influenced their real weight loss. RESULTS: Between 2016 and 2019, 190 patients underwent Roux-en-Y gastric bypass. Of these patients, 36 were lost to follow-up and 154 were included in this study. At 24 months, 16 patients had surgical nonresponse, defined as a real weight of +1 standard deviation higher than that predicted. Among these patients, 44% had a weight of ≥+1 standard deviation higher than predicted at 3 months, and 63% at 12 months. The positive and negative predictive values at 12 months were 59% and 95%, respectively. Patients with a lower hypothetically wanted weight (38.5%) exhibited greater weight loss (P < .05). CONCLUSIONS: The predictive weight loss model of Seyssel et al. enables rapid detection of surgical nonresponse, allowing physicians to react as early as 3 months postsurgery. Patients' overestimation of postoperative weight loss was positively correlated with the actual weight loss achieved.

Building 3D Generative Models from Minimal Data.

We propose a method for constructing generative models of 3D objects from a single 3D mesh and improving them through unsupervised low-shot learning from 2D images. Our method produces a 3D morphable model that represents shape and albedo in terms of Gaussian processes. Whereas previous approaches have typically built 3D morphable models from multiple high-quality 3D scans through principal component analysis, we build 3D morphable models from a single scan or template. As we demonstrate in the face domain, these models can be used to infer 3D reconstructions from 2D data (inverse graphics) or 3D data (registration). Specifically, we show that our approach can be used to perform face recognition using only a single 3D template (one scan total, not one per person). We extend our model to a preliminary unsupervised learning framework that enables the learning of the distribution of 3D faces using one 3D template and a small number of 2D images. Our approach is motivated as a potential model for the origins of face perception in human infants, who appear to start with an innate face template and subsequently develop a flexible system for perceiving the 3D structure of any novel face from experience with only 2D images of a relatively small number of familiar faces.

The ultrastructure of the apical organ of Curini-Galletti's larva, a new polyclad larval type.

Polycladida are the only free-living flatworms with a planktonic larval stage in some species. Currently, it is not clear if a larval stage is ancestral in polyclads, and which type of larva that would be. Known polyclad larvae are Müller's larva, Kato's larva and Goette's larva, differing by body shape and the number of lobes and eyes. A valuable character for the comparison and characterisation of polyclad larval types is the ultrastructural composition of the apical organ. This organ is situated at the anterior pole of the larva and is associated with at least one ciliary tuft. The larval apical organ of Theama mediterranea features two multiciliated apical tuft sensory cells. Six unfurcated apical tuft gland cell necks are sandwiched between the apical tuft sensory cells and two anchor cells and have their cell bodies located lateral to the brain. Another type of apical gland cell necks is embedded in the anchor cells. Ventral to the apical tuft, ciliated sensory neurons are present, which are neighbouring the cell necks of two furcated apical tuft gland cells. Based on the ultrastructural organisation of the apical organ and other morphological features, like a laterally flattened wedge-shaped body and three very small lobes, we recognise the larva of T. mediterranea as a new larval type, which we name Curini-Galletti's larva after its first discoverer. The ultrastructural similarities of the apical organ in different polyclad larvae support their possible homology, that is, all polyclad larvae have likely evolved from a common larva.

SARS-CoV-2 vaccination in inflammatory bowel disease patients is not associated with flares: a retrospective single-centre Swiss study.

INTRODUCTION: Vaccination hesitancy is an important barrier to vaccination among IBD patients. The development of adverse events is the main concern reported. The purpose of this monocentric study was to assess SARS-CoV-2 vaccination safety in IBD patients by evaluating the postvaccination flare risk and incidence of overall adverse events. METHODS: Surveys were handed out on three consecutive months to each patient presenting at the Crohn-Colitis Centre, where they documented their vaccination status and any side effects experienced after vaccination.Dates of flares occurring in 2021 were recorded from their electronic medical records. Baseline and IBD characteristics and flare incidence were compared between the vaccinated and unvaccinated patients, and among the vaccinated population before and after their vaccination doses. The characteristics of patients who developed side effects and of those who did not were compared. RESULTS: We enrolled 396 IBD patients, of whom 91% were vaccinated. The proportion of patients who experienced flares was statistically not different between the vaccinated and the unvaccinated population (1.8 vs 2.6 flares per 100 person-months (p = 0.28)). Among vaccinated patients, there was no difference across the prevaccination, 1 month post any vaccination, and more than 1 month after any vaccination periods, and between the Spikevax and Cominarty subgroups. Overall, 46% of patients reported vaccination side effects, mostly mild flu-like symptoms. CONCLUSION: SARS-CoV-2 vaccination with mRNA vaccines seems safe, with mostly mild side effects. The IBD flare risk is not increased in the month following any vaccination.

Dehydrated human amnion/chorion membrane allograft with spongy layer to significantly improve the outcome of chronic non-healing wounds.

We investigated the healing effect of a new dehydrated amnion/chorion membrane with a spongy layer over a 30-month period in 32 patients with 53 chronic non-healing wounds of different aetiologies. Wounds with <40% surface reduction after 4 weeks of best wound treatment underwent weekly allograft application by a certified wound specialist based on national guidelines and a standardised protocol until complete healing or definite treatment interruption. The main outcome measure was the percentage of wound surface reduction from baseline calculated using digital planimetry follow-up photographs. Overall, 38 (71.7%) wounds presented a favourable outcome (70%-100% area reduction), with 35 (66%) completely healing over a median time of 77 days (range 29-350 days). Favourable outcomes were observed in 75% of traumatic wounds, surgical wounds, venous leg ulcers and pressure injuries, as well as in 50% of ischaemic wounds. Wounds being present <12 months were significantly more likely to have a favourable outcome than more long-standing wounds (χ2 = 7.799; p = 0.005; OR = 3.378; 95% CI, 1.410-8.092). Thus, treatment with dehydrated amnion/chorion membrane with a spongy layer improves the outcome of non-healing wounds of different aetiologies and, therefore, has to be considered early in the management of refractory wounds.

Convergent evolution of the sensory pits in and within flatworms.

BACKGROUND: Unlike most free-living platyhelminths, catenulids, the sister group to all remaining flatworms, do not have eyes. Instead, the most prominent sensory structures in their heads are statocysts or sensory pits. The latter, found in the family Stenostomidae, are concave depressions located laterally on the head that represent one of the taxonomically important traits of the family. In the past, the sensory pits of flatworms have been homologized with the cephalic organs of nemerteans, a clade that occupies a sister position to platyhelminths in some recent phylogenies. To test for this homology, we studied morphology and gene expression in the sensory pits of the catenulid Stenostomum brevipharyngium. RESULTS: We used confocal and electron microscopy to investigate the detailed morphology of the sensory pits, as well as their formation during regeneration and asexual reproduction. The most prevalent cell type within the organ is epidermally-derived neuron-like cells that have cell bodies embedded deeply in the brain lobes and long neurite-like processes extending to the bottom of the pit. Those elongated processes are adorned with extensive microvillar projections that fill up the cavity of the pit, but cilia are not associated with the sensory pit. We also studied the expression patterns of some of the transcription factors expressed in the nemertean cephalic organs during the development of the pits. Only a single gene, pax4/6, is expressed in both the cerebral organs of nemerteans and sensory pits of S. brevipharyngium, challenging the idea of their deep homology. CONCLUSIONS: Since there is no morphological or molecular correspondence between the sensory pits of Stenostomum and the cerebral organs of nemerteans, we reject their homology. Interestingly, the major cell type contributing to the sensory pits of stenostomids shows ultrastructural similarities to the rhabdomeric photoreceptors of other flatworms and expresses ortholog of the gene pax4/6, the pan-bilaterian master regulator of eye development. We suggest that the sensory pits of stenostomids might have evolved from the ancestral rhabdomeric photoreceptors that lost their photosensitivity and evolved secondary function. The mapping of head sensory structures on plathelminth phylogeny indicates that sensory pit-like organs evolved many times independently in flatworms.

Evolutionary dynamics of whole-body regeneration across planarian flatworms.

Regenerative abilities vary dramatically across animals. Even amongst planarian flatworms, well-known for complete regeneration from tiny body fragments, some species have restricted regeneration abilities while others are almost entirely regeneration incompetent. Here, we assemble a diverse live collection of 40 planarian species to probe the evolution of head regeneration in the group. Combining quantification of species-specific head-regeneration abilities with a comprehensive transcriptome-based phylogeny reconstruction, we show multiple independent transitions between robust whole-body regeneration and restricted regeneration in freshwater species. RNA-mediated genetic interference inhibition of canonical Wnt signalling in RNA-mediated genetic interference-sensitive species bypassed all head-regeneration defects, suggesting that the Wnt pathway is linked to the emergence of planarian regeneration defects. Our finding that Wnt signalling has multiple roles in the reproductive system of the model species Schmidtea mediterranea raises the possibility that a trade-off between egg-laying, asexual reproduction by fission/regeneration and Wnt signalling drives regenerative trait evolution. Although quantitative comparisons of Wnt signalling levels, yolk content and reproductive strategy across our species collection remained inconclusive, they revealed divergent Wnt signalling roles in the reproductive system of planarians. Altogether, our study establishes planarians as a model taxon for comparative regeneration research and presents a framework for the mechanistic evolution of regenerative abilities.

BOSS: Bones, organs and skin shape model.

A virtual anatomical model of a patient can be a valuable tool for enhancing clinical tasks such as workflow automation, patient-specific X-ray dose optimization, markerless tracking, positioning, and navigation assistance in image-guided interventions. For these tasks, it is highly desirable that the patient's surface and internal organs are of high quality for any pose and shape estimate. At present, the majority of statistical shape models (SSMs) are restricted to a small number of organs or bones or do not adequately represent the general population. To address this, we propose a deformable human shape and pose model that combines skin, internal organs, and bones, learned from CT images. By modeling the statistical variations in a pose-normalized space using probabilistic PCA while also preserving joint kinematics, our approach offers a holistic representation of the body that can be beneficial for automation in various medical applications. In an interventional setup, our model could, for example, facilitate automatic system/patient positioning, organ-specific iso-centering, automated collimation or collision prediction. We assessed our model's performance on a registered dataset, utilizing the unified shape space, and noted an average error of 3.6 mm for bones and 8.8 mm for organs. By utilizing solely skin surface data or patient metadata like height and weight, we find that the overall combined error for bone-organ measurement is 8.68 mm and 8.11 mm, respectively. To further verify our findings, we conducted additional tests on publicly available datasets with multi-part segmentations, which confirmed the effectiveness of our model. In the diverse TotalSegmentator dataset, the errors for bones and organs are observed to be 5.10mm and 8.72mm, respectively. Our work shows that anatomically parameterized statistical shape models can be created accurately and in a computationally efficient manner. The proposed approach enables the construction of shape models that can be directly integrated into to various medical applications.

Small intestinal perforation secondary to metastasis from skin squamous cell carcinoma: A case report and literature review.

INTRODUCTION AND IMPORTANCE: Primary and metastatic carcinoma of the small intestine are rare. While most of these malignancies are adenocarcinomas, squamous cell carcinoma (SCC) of the gastrointestinal tract is uncommon. We present a case report of a rare occurrence of skin SCC metastasizing to the ileum, highlighting diagnostic challenges and clinical implications. CASE PRESENTATION: An 83-year-old female had a history of cutaneous SCC excision in the right temporal region two years prior to the current emergency department visit, followed by metastatic recurrence in a right intra-parotid lymph node treated with radiotherapy. The patient exhibited septic shock and an acute abdomen, and an abdominal computed tomography scan revealed signs of intestinal perforation. Emergency exploratory laparotomy confirmed purulent peritonitis and perforation of the terminal ileum. Subsequently, a 20 cm intestinal resection was performed. Histopathological examination of the resected specimen revealed a 4 cm perforated SCC of the small intestine (pT4 pN0 L0 V1 Pn0 R0). CLINICAL DISCUSSION: Metastases of the small intestine are rare. The primary sites for these metastases are typically the uterus, cervix, colon, lung, breast, or melanoma. SCC of the small intestine is particularly rare and poses challenges in diagnosis owing to non-specific symptoms. The prognosis for SCC of the small intestine is generally poor, and the potentially aggressive behavior of some skin SCC emphasizes the need for increased awareness and vigilance in managing such cases. CONCLUSION: This case report underscores the importance of considering metastatic disease in the small bowel of patients with a history of skin SCC who present with new-onset abdominal symptoms.

Retrograde mesenteric perfusion from the deep femoral artery in a patient with a recurrent anastomotic aneurysm in the groin: a case report.

In patients with atherosclerotic disease in two of the three main vessels in the splanchnic circulation, symptoms of chronic mesenteric ischemia may arise, depending on the disease chronicity and the presence of mesenteric collateral pathways. The most commonly described collateral pathways are between the celiac artery (CA), superior mesenteric artery (SMA), and inferior mesenteric artery (IMA); and between the IMA and the internal iliac artery (IIA). Another collateral network between the deep femoral artery and the IIA can also become important, especially in patients with aorto-iliac occlusion. Here we report a patient with a symptomatic anastomotic aneurysm of the right femoral artery after a previous aorto-bi-femoral bypass. This patient's bowel viability relied on a well-developed collateral network from the ipsilateral deep femoral artery. This unusual anatomy required special surgical considerations and planning, to minimize the risk of perioperative mesenteric ischemia. During open repair, distal femoral debranching with a distal-to-proximal anastomotic sequence allowed minimizing of the ischemic time, and avoidance of potential ischemic complications from the visceral circulation. This case emphasizes the importance and benefit of the deep femoral artery and its collaterals as a reserve network of the splanchnic circulation. Favorable outcomes can be achieved with careful analysis of the preoperative imaging and proper planning, with adaptation of the surgical strategy.

A new species of Cycloporus from the Adriatic Sea, with an updated phylogeny of the families Euryleptidae and Stylostomidae (Polycladida, Platyhelminthes).

We describe Cycloporus pinkipus sp. n., a new polyclad flatworm species from the Adriatic coast of Croatia using live images, histological sections, and a molecular marker. It is the fifteenth described species of Cycloporus Lang, 1884 and the second described congener in the Mediterranean. The genus Cycloporus is characterised by a small oval body, tentacular bumps and the name-giving marginal pores. Cycloporus pinkipus sp. n. has a smooth dorsal surface, which is transparent creamy white with light brown to yellow spots, covered with prominent serial pink spots on the inner rim of the body margin. There is little variation of the genital organs between different species of the genus, therefore we recognise C. pinkipus sp. n. as a new species in particular due to its unique coloration, and a unique partial large nuclear ribosomal subunit (28S) sequence. In recent years the family Euryleptidae Stimpson, 1857, which also contains the genus Cycloporus, was discussed and revised in several molecular studies. In an updated molecular phylogeny of the Polycladida based on partial 18S and 28S rDNA marker genes, C. pinkipus sp. n. was recovered in a clade of many other Cycloporus species within Euryleptidae.

Cooperation, coordination, or collaboration? A structured review of buyers' partnerships to support sustainable sourcing in supply chains.

The growing need of today's global supply chains for a paradigm shift to sustainability requires all supply chain members to establish partnership relationships. However, the existing literature fails to provide a comprehensive understanding of these partnerships. The present research contributes to understanding the nature and structure of the buyers' partnerships for enhancing sustainable sourcing. A structured review methodology is adopted to collect information about supply chain partnerships from the literature on sustainable sourcing. Then, a content analysis is conducted on the collected information using a comprehensive partnership framework, i.e., the McNamara framework. The framework suggests ten interconnected facets to characterize the structure of a partnership and classifies the nature of partnerships into three categories called cooperation, coordination, and collaboration. The findings indicate that cooperative partnerships are not effective for promoting sustainable sourcing as they lack the exchange of resources between the participating organizations. In contrast, coordinative partnerships are mainly effective for tactical and operational initiatives to address reactive, end-of-pipe solutions for sustainable sourcing. Finally, collaborative partnerships should be mainly initiated at the strategic level to design proactive solutions for sustainable sourcing. Some practical implications are provided to facilitate the transition of supply chains to sustainability. Some open questions for future research are also provided.

The ultrastructure of the apical organ of the Müller's larva of the tiger flatworm Prostheceraeus crozieri.

The tiger flatworm Prostheceraeus crozieri (Polycladida) develops via an eight-lobed, and three-eyed planktonic Müller's larva. This larva has an apical organ, ultrastructural details of which remain elusive due to a scarcity of studies. The evolution and possible homology of the polyclad larva with other spiralian larvae is still controversial. Here, we provide ultrastructural data and three-dimensional reconstructions of the apical organ of P. crozieri. The apical organ consists of an apical tuft complex and a dorso-apical tuft complex. The apical tuft complex features a central tuft of five long cilia, which emerge from four or five individual cells that are themselves encircled by two anchor cells. The necks of six multibranched gland cells are sandwiched between ciliated tuft cell bodies and anchor cells. The proximal parts of the ciliated cell bodies are in contact with the lateral brain neuropil via gap junctions. Located dorsally of the apical tuft complex, the dorso-apical tuft complex is characterized by several long cilia of sensory neurons, these emerge from an epidermal lumen and are closely associated with several gland cells that form a crescent apically around the dorsal anchor cell, and laterally touch the brain neuropil. Such ciliated sensory neurons emerging from a ciliated lumen are reminiscent of ampullary cells of mollusc and annelid larvae; a similar cell type can be found in the hoplonemertean decidula larva. We hypothesize that the ampullary-like cells and the tuft-forming sensory cells in the apical organs of these spiralian larvae could be homologous.

Length of the Remnant Cystic Duct and Bile Duct Stone Recurrence: a Case‒Control Study.

BACKGROUND: Since the introduction of the Critical View of Safety approach in laparoscopic cholecystectomy, exposure of the common bile duct, and common hepatic duct is not recommended, therefore, the length of the cystic duct remnant is no longer controlled. The aim of this case‒control study is to evaluate the relationship between the length of the cystic duct remnant and the risk for bile duct stone recurrence after cholecystectomy. METHODS: All MRIs with dedicated sequences of the biliary tract taken between 2010 and 2020 from patients who underwent prior cholecystectomy were reviewed. The length of the cystic duct remnant was measured and compared between the patients with and without bile duct stones using multivariate logistic regression analysis. RESULTS: A total of 362 patients were included in this study, 23.5% of whom had bile duct stones on MRI. The cystic duct remnant was significantly longer in the patients with stones than in the control group (median 31 mm versus 18 mm, P < 0.001). In the MRIs performed > 2 years after cholecystectomy, the cystic duct remnant was also significantly longer in the patients with bile duct stones (median 32 mm versus 21 mm, P < 0.001). A cystic duct remnant ≥ 15 mm in length increased the odds of stones (OR = 2.3, P = 0.001). Overall, the odds of bile duct stones increased with an increasing cystic duct remnant length (≥ 45 mm, OR = 5.0, P < 0.001). CONCLUSIONS: An excessive cystic duct remnant length increases the odds of recurrent bile duct stones after cholecystectomy.

A phylogenomic approach to resolving interrelationships of polyclad flatworms, with implications for life-history evolution.

Platyhelminthes (flatworms) are a diverse invertebrate phylum useful for exploring life-history evolution. Within Platyhelminthes, only two clades develop through a larval stage: free-living polyclads and parasitic neodermatans. Neodermatan larvae are considered evolutionarily derived, whereas polyclad larvae are hypothesized to be ancestral due to ciliary band similarities among polyclad and other spiralian larvae. However, larval evolution has been challenging to investigate within polyclads due to low support for deeper phylogenetic relationships. To investigate polyclad life-history evolution, we generated transcriptomic data for 21 species of polyclads to build a well-supported phylogeny for the group. The resulting tree provides strong support for deeper nodes, and we recover a new monophyletic clade of early branching cotyleans. We then used ancestral state reconstructions to investigate ancestral modes of development within Polycladida and more broadly within flatworms. In polyclads, we were unable to reconstruct the ancestral state of deeper nodes with significant support because early branching clades show diverse modes of development. This suggests a complex history of larval evolution in polyclads that likely includes multiple losses and/or multiple gains. However, our ancestral state reconstruction across a previously published platyhelminth phylogeny supports a direct developing prorhynchid/polyclad ancestor, which suggests that a larval stage in the life cycle evolved along the polyclad stem lineage or within polyclads.

Limitations and prospects in the management of IPMN: a retrospective, single-center observational study.

BACKGROUND: With increasing use and enhanced accuracy of cross-sectional imaging, the diagnosis of intraductal papillary mucinous neoplasms (IPMNs) of the pancreas has increased over the last few decades. The extent to which malignant transformation occurs remains unclear, making the management of IPMNs controversial. The aim of this study was to evaluate the progression rate and outcome of follow-up in patients with IPMNs. METHODS: A database of all patients diagnosed with IPMN at the Cantonal Hospital HFR Fribourg, Switzerland, between January 2006 and December 2019 with a follow-up of at least 6 months was analyzed retrospectively. Descriptive statistics were performed on patient demographics, IPMN characteristics, and follow-up data. RESULTS: A total of 56 patients were included in this study. Ten patients underwent primary surgery, 46 were enrolled in a surveillance program.21.7% (n = 5) of patients under surveillance presented with worrisome features of IPMN; progression rates were significantly higher in these patients (p = 0.043). Most progression occurred in the early follow-up period. Five patients underwent surgery due to progression, of which 2 presented high-grade dysplasia and 2 malignancy on postoperative histology. CONCLUSIONS: The limited predictive value of current guidelines may lead to surgical overtreatment, and the decision to proceed with surgical resection should be made with caution. Further prospective analyses and the development of novel biomarkers are needed to better understand the natural history of IPMN and improve diagnostic precision.

Do Not Lose Your Head Over the Unequal Regeneration Capacity in Prolecithophoran Flatworms.

One of the central questions in studying the evolution of regeneration in flatworms remains whether the ancestral flatworm was able to regenerate all body parts, including the head. If so, this ability was subsequently lost in most existent flatworms. The alternative hypothesis is that head regeneration has evolved within flatworms, possibly several times independently. In the well-studied flatworm taxon Tricladida (planarians), most species are able to regenerate a head. Little is known about the regeneration capacity of the closest relatives of Tricladida: Fecampiida and Prolecithophora. Here, we analysed the regeneration capacity of three prolecithophoran families: Pseudostomidae, Plagiostomidae, and Protomonotresidae. The regeneration capacity of prolecithophorans varies considerably between families, which is likely related to the remaining body size of the regenerates. While all studied prolecithophoran species were able to regenerate a tail-shaped posterior end, only some Pseudostomidae could regenerate a part of the pharynx and pharynx pouch. Some Plagiostomidae could regenerate a head including the brain and eyes, provided the roots of the brain were present. The broad spectrum of regeneration capacity in Prolecithophora suggests that head regeneration capacity is not an apomorphy of Adiaphanida.

Sticking Together an Updated Model for Temporary Adhesion.

Non-parasitic flatworms are known to temporarily attach to the substrate by secreting a multicomponent bioadhesive to counteract water movements. However, to date, only species of two higher-level flatworm taxa (Macrostomorpha and Proseriata) have been investigated for their adhesive proteins. Remarkably, the surface-binding protein is not conserved between flatworm taxa. In this study, we sequenced and assembled a draft genome, as well as a transcriptome, and generated a tail-specific positional RNA sequencing dataset of the polyclad Theama mediterranea. This led to the identification of 15 candidate genes potentially involved in temporary adhesion. Using in situ hybridisation and RNA interference, we determined their expression and function. Of these 15 genes, 4 are homologues of adhesion-related genes found in other flatworms. With this work, we provide two novel key components on the flatworm temporary adhesion system. First, we identified a Kringle-domain-containing protein (Tmed-krg1), which was expressed exclusively in the anchor cell. This in silico predicted membrane-bound Tmed-krg1 could potentially bind to the cohesive protein, and a knockdown led to a non-adhesive phenotype. Secondly, a secreted tyrosinase (Tmed-tyr1) was identified, which might crosslink the adhesive proteins. Overall, our findings will contribute to the future development of reversible synthetic glues with desirable properties for medical and industrial applications.

Optimized Isolation and Characterization of C57BL/6 Mouse Hepatic Stellate Cells.

To obtain meaningful results of hepatic stellate cell (HSC) function, it is crucial to use highly pure HSC populations. Our aim was to optimize HSC isolation from mice livers without exploiting the characteristically transient vitamin A autofluorescence of HSC. HSCs were isolated from C57BL/6 mice using a two-step collagenase digestion and Nycodenz gradient separation followed by CD11b-negative sorting step in order to remove contaminating macrophages and dendritic cells. Isolated cells were analyzed for yield, viability, purity, and potential new markers using immunofluorescence and flow cytometry. We obtained a yield of 350,595 ± 100,773 HSC per mouse liver and a viability of isolated cells of 92.4 ± 3.1%. We observed a low macrophage/dendritic cell contamination of 1.22 ± 0.54%. Using flow cytometry, we demonstrated that CD38 was expressed at the surface of HSC subpopulations and that all expressed intracellular markers specific for HSC in the liver. This isolation method, avoiding fluorescent activated cell sorting (FACS), allowed isolation of HSCs with high purity. Further, flow cytometry analysis suggests that CD38 may be a reliable marker of HSCs and may include subpopulations of HSCs without retinoid droplets.