Tissue-Specific Accumulation of Orally Administered Short- and Medium-Chain Chlorinated Paraffins in Honeybees (Apis mellifera L.).

The prevalence and distribution of chlorinated paraffins (CPs) have been extensively studied in various matrices and organisms; however, there is a lack of information about insects, particularly in honeybees. To address this gap, we studied young honeybee workers exposed to short- and medium-chain CPs (SCCPs and MCCPs) at an environmentally relevant concentration of 10 mg/L for 7 days, followed by a 7-day elimination period. Results indicated that CPs could transfer into the head after oral consumption and SCCPs and MCCPs exhibited clear bioaccumulation trends: midgut > hindgut > head. An evaluation of congener group distribution patterns demonstrated that the dominant congener groups in all target tissues were C11-13Cl7-8 and C14Cl7-8 for SCCPs and MCCPs, respectively, consistent with the treated CP standards. In honeybees, a significant negative relationship was observed for the log concentration of MCCP congener groups and their log KOW, but not with their log KOA. Conversely, no such correlation was found for SCCPs. These findings suggest that honeybees have a high potential to bioaccumulate MCCPs, particularly those with a low log KOW, and exhibit weak selectivity for SCCPs.

A novel paraffin-based N/P controlled-release material for biostimulation of phenol biodegradation in groundwater.

To address the problem of the weak natural restoration ability of oligotrophic groundwater environments, a novel N/P controlled-release material (CRM) for biostimulation, prepared by an improved method, was developed. CRMs can encapsulate N and P (N/P) salts for sustained release in aquifers. Paraffin-based CRMs can be used to control N/P release rates by adjusting the particle size of CRMs and the mass ratio of the paraffin. The developed CRMs had a more remarkable adaptability to groundwater than other materials. Specifically, 0.4-cm CRMs released N/P stably and efficiently over a wide temperature range (7-25 â„ƒ), and the release properties of various CRMs were not affected by pH. The release of N/P followed Fickian diffusion, and a dissolution-diffusion model was established to elucidate the mechanism of the controlled release. In contrast to bare N/P, CRMs obviously enhanced the biodegradation rate of phenol and prolonged the effectiveness of supplying N/P. The degradation rate of phenol in the CRM system increased by 20.8 %. The different supply modes of N/P, CRMs and bare N/P, resulted in differences in salinity. Metagenomic analysis showed that this difference changed the proportion of various phenol-degrading genera and thus changed the abundance of genes associated with the phenol degradation pathway.

Parafilm® M and Strat-M® as skin simulants in in vitro permeation of dissolving microarray patches loaded with proteins.

In vitro permeation studies play a crucial role in early formulation optimisation before extensive animal model investigations. Biological membranes are typically used in these studies to mimic human skin conditions accurately. However, when focusing on protein and peptide transdermal delivery, utilising biological membranes can complicate analysis and quantification processes. This study aims to explore Parafilm®M and Strat-M® as alternatives to dermatomed porcine skin for evaluating protein delivery from dissolving microarray patch (MAP) platforms. Initially, various MAPs loaded with different model proteins (ovalbumin, bovine serum albumin and amniotic mesenchymal stem cell metabolite products) were prepared. These dissolving MAPs underwent evaluation for insertion properties and in vitro permeation profiles when combined with different membranes, dermatomed porcine skin, Parafilm®M, and Strat-M®. Insertion profiles indicated that both Parafilm®M and Strat-M® showed comparable insertion depths to dermatomed porcine skin (in range of 360-430 µm), suggesting promise as membrane substitutes for insertion studies. In in vitro permeation studies, synthetic membranes such as Parafilm®M and Strat-M® demonstrated the ability to bypass protein-derived skin interference, providing more reliable results compared to dermatomed neonatal porcine skin. Consequently, these findings present valuable tools for preliminary screening across various MAP formulations, especially in the transdermal delivery of proteins and peptides.

The effect of different doses of nonylphenol on the blood-testicular barrier integrity, hormone level, and DNA damage in the testes of rats.

Determining the molecular characteristics of the damage caused by NP exposure in the testis is very important for understanding the source of the damage and developing treatment methods accordingly. Therefore, in this study, it is aimed to evaluate the toxic effects that different doses of NP may cause in the testis, including blood-testicular barrier integrity and sperm DNA damage. For this purpose, 50 adult male Wistar albino rats were used in the study. Low, medium, and high-dose NP groups and the corn oil group were formed. After NP administration at determined doses for 15 days, the testis tissue taken under anesthesia was fixed in formaldehyde. Paraffin blocks were embedded using the routine histological tissue follow-up method. Histopathological and immunohistochemical analyses were performed by taking 5 µm thick sections from paraffin blocks. The other testicular tissue was taken for the Western blot, Elisa, and comet methods, and the findings of sperm DNA analysis and the blood-testicular barrier were examined. NP caused the seminiferous epithelium to be disorganized and have significantly fewer cells in the testes of rats in different dose NP-induced groups. Compared with the control group, mTOR, Cx43, SCF, and HSP70 protein levels were decreased, while the expression of MMP-9 levels was increased in the different NP dose groups. Furthermore, tissue testosterone and inhibin B levels and SF-1 immunoreactivity intensity gradually decreased depending on the dose increase of NP. DNA damage of testicular tissues were increased in NP groups depending on NP dose. These results suggest that it is evident that NP, a commonly used industrial chemical, is an endocrine disrupting chemical (EDC) with estrogenic activity exerting adverse effects on health and that urgent measures are needed regarding the use.

Immune activity of Ki-67 during nasal polyp development.

OBJECTIVE: Nasal polyps are non-cancerous, soft painless growth of nasal mucosa. In this study, our aim was to investigate the Ki-67 expression level in nasal polyps by immunohistochemical method. PATIENTS AND METHODS: 30 patients with nasal polyps were included in this study. Nasal polyps were processed for paraffin wax embedding protocol. Samples were fixed and embedded in paraffin blocks. 5 µm sections were stained with Hematoxylin-Eosin dye and immune stained with Ki-67 antibody. Sections were analyzed under light microscope. RESULTS: Blood parameters showed that white blood cells, hematocrit and platelet were higher than normal range. In sections of hematoxylin-eosin staining, elevated basal cells, thin basement membrane, leukocyte infiltration, collagen fibers degeneration were observed. Masson trichrome staining revealed that degenerative epithelial cells, detached basement membrane and edema were observed. Ki-67 expression was observed in mucosal epithelial cells, vascular endothelial cells and plasma cells in immune staining. CONCLUSIONS: Epithelial degeneration in nasal polyps and leukocyte infiltration induce nasal adenoma. Ki-67 expression may be a diagnostic tool for epithelial leukocyte formation.

Multiplex immunohistochemistry and high-throughput image analysis for evaluation of spatial tumor immune cell markers in human breast cancer.

BACKGROUND: The clinicopathological significance of spatial tumor-infiltrating lymphocytes (TILs) subpopulations is not well studied due to lack of high-throughput scalable methodology for studies with large human sample sizes. OBJECTIVE: Establishing a cyclic fluorescent multiplex immunohistochemistry (mIHC/IF) method coupled with computer-assisted high-throughput quantitative analysis to evaluate associations of six TIL markers (CD3, CD8, CD20, CD56, FOXP3, and PD-L1) with clinicopathological factors of breast cancer. METHODS: Our 5-plex mIHC/IF staining was shown to be reliable and highly sensitive for labeling three biomarkers per tissue section. Through repetitive cycles of 5-plex mIHC/IF staining, more than 12 biomarkers could be detected per single tissue section. Using open-source software CellProfiler, the measurement pipelines were successfully developed for high-throughput multiplex evaluation of intratumoral and stromal TILs. RESULTS: In analyses of 188 breast cancer samples from the Nashville Breast Health Study, high-grade tumors showed significantly increased intratumoral CD3+CD8+ cytotoxic T lymphocyte density (P= 0.0008, false discovery rate (FDR) adjusted P= 0.0168) and intratumoral PD-L1 expression (P= 0.0061, FDR adjusted P= 0.0602) compared with low-grade tumors. CONCLUSIONS: The high- and low-grade breast cancers exhibit differential immune responses which may have clinical significance. The multiplexed imaging quantification strategies established in this study are reliable, cost-efficient and applicable in regular laboratory settings for high-throughput tissue biomarker studies, especially retrospective and population-based studies using archived paraffin tissues.

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To understand the development mechanism of the epiphyllous bud of waterlily, we examined the morphological anatomy of the leaf-navel epiphyllous bud by paraffin section technique at four stages, and compared the differences of carbohydrate metabolism between viviparous and non-viviparous waterlily leaves. Three tropical waterlily cultivars of Brachyceras were used, including two viviparous cultivars Nymphaea 'Margaret Mary', Nymphaea 'Ruby', and a non-viviparous cultivar Nymphaea 'Pink Star'. The results showed that parenchyma cells below the epidermis of leaf-navel divided and grew continuously after the leaf unfolded, forming a closely arranged cell cluster in viviparous waterlily and raised upward to a spherical shape. In contrast, no change was observed in leaf-navel of non-viviparous waterlily with the expansion of leaves. With the development of leaves, the contents of all physiological variables except sucrose and enzyme activities in the leaves of viviparous waterlily showed a first increase and then a decrease, which was significantly higher than those of non-viviparous waterlily. The carbohydrate contents in different parts showed the order of leaf > leaf-navel > petiole (except for starch content, which was highest in the leaf-navel). The activities of sucrose synthase (SS) and acid invertase (AI) were higher than those of sucrose phosphate synthase (SPS) and neutral invertase (NI). The activities of SPS and NI in different tissues of viviparous waterlily were significantly higher than those in non-viviparous one, but SS and AI did not show pronounced cultivar advantage in viviparous cultivars. AI activity varied greatly among cultivars, whereas NI activity varied less and was at a low level in different tissues. The sucrose of Nymphaea 'Ruby' leaves was positively correlated with the SPS and AI, and significantly associated with NI. The accumulation of sucrose content increased the activities of SS and NI of waterlily leaves, which was conducive to promoting the formation of epiphyllous buds.

Immunohistochemical detection of urea transporter-A in the tear-producing part of the lacrimal system.

OBJECTIVE: Urea constitutes a physiological and presumably well-regulated constituent of tear fluid. Its lacrimal concentration is significantly decreased in dry eye disease. Urea homeostasis within the tear fluid may also depend on the expression of urea transporters. The present study reports on the expression patterns of urea transporter A (UT-A) in the cells and tissues of the ocular surface and the lacrimal glands. METHODS: UT-A immunohistochemistry was performed on 5 µm paraffin sections of paraformaldehyde-fixed human, porcine, and murine corneas, eyelids, and lacrimal glands (n = 5 each). RESULTS: UT-A immunostaining was largely comparable in all three species. UT-A signals were detected in the corneal epithelium and endothelium, in the conjunctival epithelium, in the acinar cells and excretory ducts of the lacrimal gland, Meibomian gland, and in the glands of Moll and Zeis. The Meibomian glands and the glands of Zeis exhibited a marked UT-A-positive staining in the basal cells of the alveolar epithelia and in the ductal epithelia. CONCLUSION: UT-A shows comparable expression patterns to UT-B (previous study) at the ocular surface and in the lacrimal glands, as determined by immunohistochemistry. The presence of both urea transporters in the lacrimal functional unit suggests that they are essential for the normal function of the lacrimal system and the integrity of the tear film. Potential alterations in urea transporter expression might be associated with the significant reduction of urea found in the tear fluid of dry eye patients. They may thus play an important role in the pathogenesis of dry eye disease.

Density of TMEM119-positive microglial cells in postmortem cerebrospinal fluid as a surrogate marker for assessing complex neuropathological processes in the CNS.

Routine coronal paraffin-sections through the dorsal frontal and parieto-occipital cortex of a total of sixty cases with divergent causes of death were immunohistochemically (IHC) stained with an antibody against TMEM119. Samples of cerebrospinal fluid (CSF) of the same cases were collected by suboccipital needle-puncture, subjected to centrifugation and processed as cytospin preparations stained with TMEM119. Both, cytospin preparations and sections were subjected to computer-assisted density measurements. The density of microglial TMEM119-positive cortical profiles correlated with that of cytospin results and with the density of TMEM119-positive microglial profiles in the medullary layer. There was no statistically significant correlation between the density of medullary TMEM119-positive profiles and the cytospin data. Cortical microglial cells were primarily encountered in supragranular layers I, II, and IIIa and in infragranular layers V and VI, the region of U-fibers and in circumscribed foci or spread in a diffuse manner and high density over the white matter. We have evidence that cortical microglia directly migrate into CSF without using the glympathic pathway. Microglia in the medullary layer shows a strong affinity to the adventitia of deep vessels in the myelin layer. Selected rapidly fatal cases including myocardial infarcts and drowning let us conclude that microglia in cortex and myelin layer can react rapidly and its reaction and migration is subject to pre-existing external and internal factors. Cytospin preparations proved to be a simple tool to analyze and assess complex changes in the CNS after rapid fatal damage. There is no statistically significant correlation between cytospin and postmortem interval. Therefore, the quantitative analyses of postmortem cytospins obviously reflect the neuropathology of the complete central nervous system. Cytospins provide forensic pathologists a rather simple and easy to perform method for the global assessment of CNS affliction.

GTL synthetic paraffin oil shows low liver and tissue retention compared to mineral oil.

Oral exposure to mineral oil may result in a narrow fraction of mineral oil saturated hydrocarbon (MOSH) being retained in tissues. Excess of MOSH hepatic retention may lead to the formation of lipogranuloma caused by predominantly multiring cycloalkanes (naphthenics) in a critical range of C25-C35. Although hepatic lipogranuloma is of low pathological concern, MOSH tissue deposition could be minimized by using an oil of similar quality but devoid of naphthenic structures to decrease hepatic retention. Synthetic Gas to liquid (GTL) oils offer an alternative to petroleum derived mineral oils, because they do not contain naphthenic structures. To demonstrate this point, SD rats were fed either GTL oil (99% iso-alkanes) or naphthenic mineral oil (84% cycloalkanes) at 200 mg/kg bw/day for 90 or 134 days with a recovery group. Liver, fat and mesenteric lymph nodes were analyzed for alkane sub-type levels using Online-HPLC-GC-FID and GCxGC-TOF-MS. Results indicate that at equal external dose, GTL hydrocarbons result in lower tissue levels and more rapid excretion than MOSH. GTL retained hepatic fractions were also qualitatively different than MOSH constituents. Because chemical composition differences, GTL oil show low absorption and tissue retention potential and thus an advantageous alternative to conventional mineral oil.

Short- and Medium-Chain Chlorinated Paraffins in Foods from the Sixth Chinese Total Diet Study: Occurrences and Estimates of Dietary Intakes in South China.

Food consumption has been identified as a major pathway for human exposure to short-chain chlorinated paraffins (SCCPs) and medium-chain chlorinated paraffins (MCCPs), but evaluations of SCCP and MCCP intake from major dietary sources are limited. We used the sixth Chinese Total Diet Study to perform a comprehensive investigation of SCCPs and MCCPs in cereals, vegetables, potatoes, legumes, eggs, milk, meats, and aquatic foods from nine southern provinces. The geographical distribution of CP concentrations showed higher levels in Jiangsu, Hubei, and Zhejiang provinces. The CP concentrations in most animal-origin foods were higher than those in foods of plant origin. The total estimated daily intakes (EDIs) of SCCPs and MCCPs, with average values of 7.0 × 102 and 4.7 × 102 ng kg-1 day-1, respectively, were mostly contributed by cereals, vegetables, and meats. Risk assessment indicated the EDIs of CPs posed no significant risk to residents in South China.

Evaluating age and temporal trends of chlorinated paraffins in pooled serum collected from males in Australia between 2004 and 2015.

Chlorinated paraffins (CPs) are high production volume chemicals of which some show resistance to environmental degradation, long-rang transport, bioaccumulation and toxicity potential. Information regarding their presence in humans is limited, including their human bioaccumulation potential. The present study aimed to evaluate CP levels in human serum from Australia in order to better understand their exposure and current pollution status as well as trends associated with age and time between 2004 and 2015. For this, we selected a male sub-group of the Australian population under 60 years old (n = 16 pools, total 1600 serum samples). While long-chain CP (C18-20) and most short-chain CP (C10-13, SCCPs) levels were below method detection limits (MDL), medium-chain CPs (C14-17, MCCPs) were found in most serum samples (detection frequency 94%) as well as CPs with a carbon chain length of nine (detection frequency 76%). The levels of ΣSCCPs and ΣMCCPs ranged from

Community succession in an anaerobic long-chain paraffin-degrading consortium and impact on chemical and electrical microbially influenced iron corrosion.

Community compositional changes and the corrosion of carbon steel in the presence of different electron donor and acceptor combinations were examined with a methanogenic consortium enriched for its ability to mineralize paraffins. Despite cultivation in the absence of sulfate, metagenomic analysis revealed the persistence of several sulfate-reducing bacterial taxa. Upon sulfate amendment, the consortium was able to couple C28H58 biodegradation with sulfate reduction. Comparative analysis suggested that Desulforhabdus and/or Desulfovibrio likely supplanted methanogens as syntrophic partners needed for C28H58 mineralization. Further enrichment in the absence of a paraffin revealed that the consortium could also utilize carbon steel as a source of electrons. The severity of both general and localized corrosion increased in the presence of sulfate, regardless of the electron donor utilized. With carbon steel as an electron donor, Desulfobulbus dominated in the consortium and electrons from iron accounted for ∼92% of that required for sulfate reduction. An isolated Desulfovibrio spp. was able to extract electrons from iron and accelerate corrosion. Thus, hydrogenotrophic partner microorganisms required for syntrophic paraffin metabolism can be readily substituted depending on the availability of an external electron acceptor and a single paraffin-degrading consortium harbored microbes capable of both chemical and electrical microbially influenced iron corrosion.

Phenotypic Adaptations Help Rhodococcus erythropolis Cells during the Degradation of Paraffin Wax.

During crude oil extraction, the reduction in temperature and pressure results in the precipitation of paraffin wax that contains 20-40 carbon chain hydrocarbons. The paraffin wax may accumulate inside production tubes, pipelines, and processing facilities, and also in tankers during petroleum transportation. There are few bacterial strains that are able to degrade solid substrates. In the present study, the biodegradation of paraffin is evaluated using Rhodococcus erythropolis cells. This bacterium is able to grow using paraffin wax from an oil refinery plant as the sole carbon source. The cells grow as a thick biofilm over the solid substrate, make scale-like structures that increase the area of the initially smooth surface of paraffin, produce biosurfactants, and become more negatively charged than ethanol- or glucose-grown cells. When paraffin wax is supplied as microparticles, to increase the cell-substrate contact area and to simulate paraffin precipitation, the cells also adjust the composition of the fatty acids of the phospholipids of the cellular membrane to decrease its fluidity and paraffin biodegradation increases considerably. The study suggests that the phenotypic adaptation of R. erythropolis cells may be used to degrade paraffin wax under real conditions.

Bioaccumulation and biomagnification of short-chain chlorinated paraffins in marine organisms from the Pearl River Estuary, South China.

Short-chain chlorinated paraffins (SCCPs) are a series of new persistent organic pollutants, posing a risk of significant adverse effects to biota. Increasing attention has been paid to SCCP pollution in China as large amounts of chlorinated paraffin (CP) products containing SCCPs have been produced and used there. However, knowledge of the bioaccumulation of SCCPs in marine organisms from the Pearl River Estuary (PRE), Southern China, is still scarce. In this study, SCCP concentrations were measured in seawater, sediments, and marine organisms from the PRE. SCCP concentrations ranged from 180 to 460 ng/L in seawater, from 180 to 620 ng/g dry weight (dw) in sediments, and from 870 to 36,000 ng/g lipid weight (lw) in marine biota samples. C10-11 SCCPs were the predominant homologues in all the samples, with an average abundance of 68% in seawater, 57% in sediments, and 56-77% in marine organisms. However, chlorine patterns of SCCPs in seawater, sediments, and marine organisms were different. Cl8-10 SCCPs dominated in sediments, whereas Cl5-7 SCCPs were the predominant SCCP homologues in water and most organism species. The logarithm bioaccumulation factors (BAFs) of SCCPs ranged from 1.6 to 3.0, and increased significantly with the increase of Kow values for most marine biota species, indicating that Kow was the major factor controlling the bioaccumulation of SCCPs and that SCCPs with higher lipophilicity were more prone to being bioaccumulated from water. Opposite to that observed for log BAFs, biota-sediment accumulation factors of specific SCCPs (range: 0.01-30) decreased significantly with the increase of Kow values. The biomagnification factor of total SCCPs for oyster-mangrove crab was 2.40, implying the potential biomagnification of SCCPs for benthos in the PRE.

Bioaccessibility of short chain chlorinated paraffins in meat and seafood.

Bioaccessibility of short chain chlorinated paraffins (SCCPs), which is important for estimation of dietary exposure, has not been evaluated in previous studies. In the present study, we determined the bioaccessibility of SCCPs in meat (pork, beef and chicken) and seafood (fish, clams, and prawns) using the colon-extended physiologically based extraction test as an in vitro model. The bioaccessibility percentages (BAs) ranged from 33% to 84% in the fed state and from 41% to 63% in the unfed state. The BAs observed in the fed state were lower than in the unfed state in most samples, except for pork sample, which had the highest lipid content. This could be attributed to the effects caused by dietary components added in the fed state. The effects of lipid and protein in samples on bioaccessibility were investigated. In food with a high lipid content, like pork in this study, lipid was the main factor controlling SCCP bioaccessibility. In the other five foods, which had low-medium lipid contents, BA in the unfed state was positively correlated with lipid content (p < 0.05) and negatively correlated with the protein-to-lipid content ratio (p < 0.05). No significant relationships between lipid and protein content and BA were found in the fed state. As to SCCP congener groups, a positive relationship between the BAs of SCCP congener groups and their octanol-water partition coefficients (log Kow) was found in pork sample in the fed state (p < 0.05). The BAs obtained in samples from fish, prawn, clam, and pork in the unfed state and that obtained in clam sample in the fed state were negatively correlated with log Kow (p < 0.05). We calculated more accurate estimated dietary intakes of SCCPs using our SCCP bioaccessibility data. These results will contribute to more reliable dietary risk assessments of SCCPs.

Validation of a HRGC-ECNI/LRMS method to monitor short-chain chlorinated paraffins in human plasma.

Short-chain chlorinated paraffins (SCCPs) are produced in high volume and have the high potential to pose a threat to human health. However, little information is available for SCCP contamination in human blood/plasma/serum, mainly due to the difficulty of sample preparation and quantitative analysis. A method using high resolution gas chromatography coupled with electron capture negative ionization low resolution mass spectrometry (HRGC-ECNI/LRMS) was developed and validated to measure SCCPs in human plasma. The pretreatment process included protein denaturation and lipid elimination, liquid-liquid extraction with a mixture of n-hexane/dichloromethane (1:1, V/V), and cleanup on a multi-layer silica column. The blank controls, including procedural blank, vacuum blood collection tube blank, and instrumental blank, were the most pivotal points for the reliable analysis of SCCPs. The average value of procedural blanks was 9.0ng/g; and the method detection limit (MDL), calculated as the sum of the average procedural blank value and 3 times of the standard deviation of the procedural blanks, was 12.6ng/g plasma. The validated method was applied to measure the concentrations of the total SCCPs (∑SCCPs) in 50 plasma samples from a general population. The measured plasma concentrations of ∑SCCPs ranged from

Isolation and characterization of biosurfactant producing and oil degrading Bacillus subtilis MG495086 from formation water of Assam oil reservoir and its suitability for enhanced oil recovery.

The strains isolated from the formation water were characterized and screened considering their crude oil degradation capability and biosurfactant production ability. The growth kinetics study of isolated Bacillus subtilis MG495086 was carried out by varying growth parameters i.e. carbon source, temperature, pH and salinity. The biosurfactant production was optimized adopting RSM-CCD considering carbon source (1-5%), pH (3-11) and temperature (25-65 °C) as matrix parameters. The optimum biosurfactant production (6.3 ±â€¯0.1 g/L) and the minimum surface tension 29.85 mN/m were obtained after 96 h of incubation under optimal conditions i.e. 3.8% (v/v) of light-paraffin oil as sole carbon source at 62.4 °C and pH 7.7 with the maximum oil degradation capability of 91.3 ±â€¯5%. Critical micelle concentration value of crude biosurfactant was found to be 40 mg/L with high emulsification activity of 72.45 ±â€¯0.85%. The produced biosurfactant was identified as lipopeptide (Surfactin) and characterized using various analytical techniques to establish its suitability for microbial enhanced oil recovery.

Pancreatic Islet Embedding for Paraffin Sections.

Experiments using isolated pancreatic islets are important for diabetes research, but islets are expensive and of limited abundance. Islets contain a mixed cell population in a structured architecture that impacts function, and human islets are widely variable in cell type composition. Current frequently used methods to study cultured islets include molecular studies performed on whole islets, lumping disparate islet cell types together, or microscopy or molecular studies on dispersed islet cells, disrupting islet architecture. For in vivo islet studies, paraffin-embedded pancreas sectioning is a powerful technique to assess cell-specific outcomes in the native pancreatic environment. Studying post-culture islets by paraffin sectioning would offer several advantages: detection of multiple outcomes on the same islets (potentially even the exact-same islets, using serial sections), cell-type-specific measurements, and maintaining native islet cell-cell and cell-substratum interactions both during experimental exposure and for analysis. However, existing techniques for embedding isolated islets post-culture are inefficient, time consuming, prone to loss of material, and generally produce sections with inadequate islet numbers to be useful for quantifying outcomes. Clinical pathology laboratory cell block preparation facilities are inaccessible and impractical for basic research laboratories. We have developed an improved, simplified bench-top method that generates sections with robust yield and distribution of islets. Fixed islets are resuspended in warm histological agarose gel and pipetted into a flat disc on a standard glass slide, such that the islets are distributed in a plane. After standard dehydration and embedding, multiple (10+) 4 - 5 µm sections can be cut from the same islet block. Using this method, histological and immunofluorescent analyses can be performed on mouse, rat, and human islets. This is an effective, inexpensive, time-saving approach to assess cell-type-specific, intact-architecture outcomes from cultured islets.

Bioaccumulation of organic pollutants in Indo-Pacific humpback dolphin: A review on current knowledge and future prospects.

Indo-Pacific humpback dolphin (Sousa chinensis) are chronically exposed to organic pollutants since they inhabit shallow coastal waters that are often impacted by anthropogenic activities. The aim of this review was to evaluate existing knowledge on the occurrence of organic pollutants in Indo-Pacific humpback dolphins, identify knowledge gaps, and offer recommendations for future research directions. We discussed the trends in the bioaccumulation of organic pollutants in Indo-Pacific humpback dolphins focusing on sources, physicochemical properties, and usage patterns. Furthermore, we examined factors that influence bioaccumulation such as gender, age, dietary intake and tissue-specific distribution. Studies on bioaccumulation in Indo-Pacific humpback dolphin remain scarce, despite high concentrations above 13,000 ng/g lw we previously detected for PFOS, ∑PBDE and chlorinated paraffins. The maximum concentration of organochlorines detected was 157,000 ng/g wt. Furthermore, variations in bioaccumulation were shown to be caused by factors such as usage patterns and physicochemical properties of the pollutant. However, restrictions in sampling inhibit investigations on exposure pathway and toxicity of organic pollutants in Indo-Pacific humpback dolphin. We proposed the use of biopsy sampling, predictive bioaccumulation and toxicity modeling, and monitoring other emerging contaminants such as microplastics and pharmaceuticals for future health risk assessment on this critically endangered marine mammal species.