Heavy metal levels and human health risk implications associated with fish consumption from the lower Omo river (Lotic) and Omo delta lake (Lentic), Ethiopia.

This study is the first to determine the levels of heavy metals in commercially important fish species, namely Lates niloticus and Oreochromis niloticus and the potential human health risks associated with their consumption. A total of 120 fish samples were collected from the lower Omo river and Omo delta, with 60 samples from each water source. The fish tissue samples (liver and muscle) were analyzed using a flame atomic absorption spectrometer for nine heavy metals (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn). The human health risk assessment tools used were the target hazard quotient (THQ), the hazard index (HI), and the target cancer risk (TCR). The mean levels of heavy metals detected in the liver and muscle of L. niloticus from the lower Omo river generally occurred in the order Fe > Zn > Pb> Cu > Mn> Cr > Co > Ni and Pb > Cu > Mn > Co > Ni, respectively. The mean levels of metals in the muscle and liver tissues of O. niloticus were in the order Fe > Pb > Zn > Mn > Cu > Cr > Co > Ni and Pb > Zn > Mn > Fe > Cu > Co > Ni, respectively. Similarly, the mean levels of heavy metals detected in the liver and muscle of L. niloticus from Omo delta occurred in the order Fe > Zn > Pb > Cu > Mn > Cr > Co > Ni and Fe > Pb > Zn > Mn > Cu > Co > Cr > Ni, respectively. The mean levels in the muscle and liver tissues of O. niloticus from the Omo delta were in the order Fe > Pb > Zn > Mn > Cu > Cr > Co > Ni and Pb > Fe > Zn > Mn > Co > Cu > Ni, respectively. The study revealed that the THQ values were below 1, indicating that consumption of L. niloticus and O. niloticus from the studied sites does not pose a potential non-carcinogenic health risk. Although the TCR values for Pb in this study were within the tolerable range, it's mean concentration in the muscle and liver tissues of both fish species from the two water bodies exceeded the permissible limit established by FAO/WHO. This is a warning sign for early intervention, and it emphasizes the need for regular monitoring of freshwater fish. Therefore, it is imperative to investigate the pollution levels and human health risks of heavy metals in fish tissues from lower Omo river and Omo delta for environmental and public health concerns.

Investigation of the Alternations in the Muscle Quality of Swimming Crab (Ovalipes punctatus) during Cold-Chain Transportation Using Physicochemical and TMT-Based Quantitative Proteomic Analysis.

Physicochemical properties and protein alterations in Ovalipes punctatus during cold-chain transportation were examined via sensory scores, water-holding capacity (WHC), glucose (GLU) content, catalase (CAT) activity, urea nitrogen (UN) content, and tandem mass tag (TMT)-based proteomic analysis. The results revealed that sensory characteristics and texture of crab muscle deteriorated during transportation. Proteomic analysis revealed 442 and 470 different expressed proteins (DEPs) in crabs after 18 h (FC) and 36 h (DC) of transportation compared with live crabs (LC). Proteins related to muscle structure and amino acid metabolism significantly changed, as evidenced by the decreased WHC and sensory scores of crab muscle. Glycolysis, calcium signaling, and peroxisome pathways were upregulated in the FC/LC comparison, aligning with the changes in GLU content and CAT activity, revealing the stress response of energy metabolism and immune response in crabs during 0-18 h of transportation. The downregulated tricarboxylic acid (TCA) cycle and carcinogenesis-reactive oxygen species pathways were correlated with the decreasing trend in CAT activity, suggesting a gradual retardation in both energy and antioxidant metabolism in crabs during 18-36 h of transportation. Furthermore, the regulated purine nucleoside metabolic and nucleoside diphosphate-related processes, with the increasing changes in UN content, revealed the accumulation of metabolites in crabs.

Analysing polypeptide antibiotics residues in animal muscle tissues: The crucial role of HRMS.

A confirmatory method for the determination of polypeptide antibiotics (bacitracin, colistin, and polymyxin B) in muscle samples has been developed. Extraction is performed with acidified methanol, and a clean-up step by solid-phase extraction with polymeric cartridges is applied. Separation by ultra-high performance liquid chromatography (UHPLC) is carried out using a solid core C18 column and gradient elution with water/acetonitrile containing 0.2% formic acid. High-resolution mass spectrometry (HRMS) (Q-Orbitrap) detection using different working modes has proved to be highly advantageous in eliminating interfering signals from endogenous matrix components. The analytical method has been successfully validated according to Commission Regulation 2021/808/EU and is currently used in a public health laboratory involved in veterinary medicines residue surveillance activities.

Towards a comprehensive understanding of the muscle proteome in Schizothorax labiatus: Insights from seasonal variations, metabolic responses, and reproductive signatures in the River Jhelum.

Proteomics is a very advanced technique used for defining correlations, compositions and activities of hundreds of proteins from organisms as well as effectively used in identifying particular proteins with varying peptide lengths and amino acid counts. In the present study, an endeavour has been put forth to create muscle proteome expression of snow trout, Schizothorax labiatus. Liquid chromatography-mass spectrometry (LC-MS) using label free quantification (LFQ) technique has extensively been carried out to explore changes in protein metabolism and its composition to discriminate across species, clarify functions and pinpoint protein biomarkers from organisms. In LFQ technique, the abundances of proteins are determined based on the signal intensities of their corresponding peptides in mass spectrometry. The main benefit of using this method is that it doesn't require pre-labelling proteins with isotopic tags, which streamlines the experimental procedure and gets rid of any bias that might have been caused by the labelling process. LFQ techniques frequently offer a wider dynamic range, making it possible to detect and quantify proteins over a broad range of abundances obtained from the complex biological materials including fish muscle. The results of proteomic analysis could provide an insight in understanding about how various proteins are expressed in response to environmental challenges. For proteomic study, two different weight groups of S. labiatus were taken from River Jhelum based on biological, physiological and logistical factors. These groups corresponded to different life stages, such as younger size and adults/brooders in order to capture potential variations in the muscle proteome related to growth and development. The proteomic analysis of S. labiatus depicted that an overall of 220 proteins in male and 228 in female fish of group 1 were noted. However, when male and female S. labiatus were examined based on spectral count and peptide abundance using ProteinLynx Global Software, a total of 10 downregulated and 32 upregulated proteins were found. In group 2 of S. labiatus, a total of 249 proteins in male and 301 in female fish were documented. When the two genders of S. labiatus were likened to one another by LFQ technique, a total of 41 downregulated and 06 upregulated proteins were identified. The variability in the protein numbers between two fish weight groups reflected biological differences, influenced by factors such as age, developmental stages, physiological condition and reproductive activities. During the study, it was observed that S. labiatus exhibited downregulated levels of proteins that were involved in feeding and growth. The contributing factors to this manifestation could be explained by lower feeding and metabolic activity of fish and decreased food availability during winter in River Jhelum. Contrarily, the fish immune response proteins were found to be significantly over-expressed in S. labiatus, indicating that the environment was more likely to undergo increased microbial infection, pollution load and anthropogenic activities. In addition, it was also discovered that there was an upregulated expression of the reproductive proteins in S. labiatus, which could be linked to the fish's pre-spawning time as the fish used in this study was collected in the winter season which is the pre-spawning period of the fish. Therefore, the present study would be useful in obtaining new insights regarding the molecular makeup of species, methods of adaptation and reactions to environmental stresses. This information contributes to our understanding of basic science and may have applications in environmental monitoring, conservation and preservation of fish species.

A non-lethal stable isotope analysis of valued freshwater predatory fish using blood and fin tissues as alternatives to muscle tissue.

Stable isotope analysis (SIA) is widely used to study trophic ecology and food webs in aquatic ecosystems. In the case of fish, muscle tissue is generally preferred for SIA, and the method is lethal in most cases. We tested whether blood and fin clips can be used as non-lethal alternatives to muscle tissue for examining the isotopic composition of two freshwater predatory fish, European catfish (Silurus glanis) and Northern pike (Esox lucius), species of high value for many freshwater systems as well as invasive species in many others. Blood samples from the caudal vein, anal fin clips, and dorsal muscle obtained by biopsy punch were collected from four catfish and pike populations (14-18 individuals per population). Subsequently, these samples were analyzed for δ13C and δ15N. The effects of alternative tissues, study site, and fish body mass on the isotopic offset were investigated. Both species showed a correlation between the isotopic offset and the tissue type, as well as the study site, but no significant relationship with the body mass. The isotopic offsets between tissues were used to calculate the conversion equations. The results demonstrated that both blood and fin clips are suitable and less invasive alternative to muscle in SIA studies focused on European catfish and Northern pike. Blood provided better correspondence to muscle isotope values. However, our results clearly demonstrated that isotopic offsets between tissues vary significantly among populations of the same species. Therefore, obtaining a muscle biopsy from several individuals in any population is advisable to gain initial insights and establish a possible population-specific inter-tissue conversion.

Evaluating the effects of graphene nanoparticles combined radio-frequency thawing on the physicochemical quality and protein conformation in hairtail (Trichiurus lepturus) dorsal muscle.

BACKGROUND: The thawing process is an essential step for a frozen marine fish. The present study aimed to investigate the effects of graphene magnetic nanoparticles combined radio-frequency thawing methods on frozen hairtail (Trichiurus lepturus) dorsal muscle. Seven thawing methods were used: air thawing, 4 °C cold storage thawing, water thawing, radio-frequency thawing (RT), radio frequency thawing combined with graphene nanoparticles (G-RT), radio frequency thawing combined with graphene oxide nanoparticles (GO-RT) and radio-frequency thawing combined with graphene magnetic nanoparticles (GM-RT). The thawing loss and centrifugal loss, electric conductivity, total volatile basic nitrogen, thiobarbituric acid reactive substances and color of thawed hairtail dorsal muscle were determined. The carbonyl content, total sulfhydryl groups, Ca2+ -ATPase activity, raman spectroscopy measurements and Fourier-transform infrared spectrometry measurements were determined using myofibrillar extracted from the dorsal muscle of hairtail. The water distribution was determined using low-field NMR techniques. RESULTS: The results demonstrated that the RT, G-RT, GO-RT and GM-RT could significantly shorten the thawing time. Moreover, GO-RT and GM-RT efficiently preserved the color of fish dorsal muscle and reduced the impact of thawing on fish quality by reducing lipid and protein oxidation. Meanwhile, the myofibrillar protein structure thawed by GO-RT and GM-RT were more stable and had a more stable secondary structure, which maintained strong systemic stability at the same time as slowing down protein oxidation. CONCLUSION: The results showed that GO-RT and GM-RT can significantly improve the thawing efficiency at the same time as effectively maintaining and improving the color and texture of thawed fish, slowing down the oxidation of proteins and lipids, and maintaining a good quality of thawed fish meat. © 2023 Society of Chemical Industry.

Non-lethal sampling method for the analysis of white muscle fatty acid profiles in European sea bass (Dicentrarchus labrax).

This study presents a novel non-lethal sampling method for assessing fatty acid (FA) composition in juvenile European sea bass (Dicentrarchus labrax) using subcutaneous white muscle biopsies. This research aimed to evaluate the suitability of the biopsy for FA analysis using two lipid extraction protocols and comparing them to a lethal routine method. The results showed that a mass of fresh tissue as low as 1.4 mg provided good quality FA chromatograms for both reserve and membrane lipids. Although the biopsy method displayed high variability in terms of FA quantity among intra-individual replicates, it showed good FA profile repeatability in both reserve and membrane lipids. The study highlights the potential of this non-lethal approach for studying FA dynamics in fish, with its application being particularly promising for ecological and experimental studies. However, careful biopsy implementation is recommended to account for potential lipid droplet and lipid distribution variability within the tissue.

Comparison of Two Total Mercury Screening and Assessment Methods in Fishes: Biopsy Punch and Dried Muscle Samples.

Rapid and effective quantification of total mercury concentrations ([THg]) in fish muscle is an important part of ongoing monitoring to provide reliable and near real-time public health guidance. Methods for quantifying THg in fish muscle frequently require the use of large sample mass and numerous preparation steps. Wet (aka fresh weight) biopsy punch samples of fish muscle have been used to quantify THg directly, without drying and homogenization. Both methods have advantages and disadvantages. We compare the use of fresh weight biopsy punches for quantifying THg to using larger, dried homogenized samples. The [THg] determination for the two sampling methods was EPA method 7473. Three separate biopsy punch samples and a large muscle sample were taken from each fish and analyzed on a Direct Mercury Analyzer. There were no statistical differences between mean log transformed wet weight [THg] from biopsy punches and homogenized muscle across all samples or within individual species. Similarly, across the range of [THg] (7.5-612.7 ng/g ww), linear regression of [THg] from biopsy punch and homogenized muscle samples was not different from a 1:1 linear relationship. Linear regression statistics of [THg] with fish fork length produced similar results for both biopsy punch and homogenized muscle samples. However, the coefficient of variation among biopsy punch replicates for individual fish was frequently above the acceptable threshold of 15%. We recommend biopsy punches be used as an effective tool for broad-scale rapid monitoring of fish resources for Hg, while homogenized muscle samples be used for fine-scale ecological and health questions.

Analysis of metabolite differences between South Korean and Chinese yellow goosefish (Lophius litulon) using capillary electrophoresis time-of-flight mass spectrometry.

The yellow goosefish is a benthic fish that belongs to the family Lophiidae and order Lophiiformes and is distributed in the Yellow and East China Seas. This study aimed to distinguish between yellow goosefish from different geographical origins by analyzing their metabolites. Capillary electrophoresis time-of-flight mass spectrometry was used to analyze metabolite profiles in the muscle tissues of yellow goosefish to distinguish between Korean and Chinese yellow goosefish. In total, 271 putative metabolites were extracted using 50% acetonitrile in water. Principal component analysis and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to distinguish different geographical origins using the metabolite profiles obtained. The R2 and Q2 values of the OPLS-DA model were 0.856 and 0.695, respectively, indicating that the model was well-fitted and had good predictability. The heat map revealed that nucleic acid and amino compounds differed between the Korean and Chinese fish, and the variable importance in the projection scores obtained from OPLS-DA showed that there were geographical differences in the primary metabolites (5'-methylthioadenosine, adenosine, uridine 5-diphosphate, guanosine 5-diphosphate, urea, homocarnosine, O-acetylcarnitine, cycloleucine, cycloleucine S-adenosylmethionine, S-adenosylhomocysteine, ethanolamine, myo-inositol 1-phosphate), which were identified as potential candidate biomarkers.

A Panel-Based Method for the Reproduction of Distinct Molecular Subtype Classifications of Muscle-Invasive Urothelial Bladder Cancer.

Transcriptome-based molecular subtype classification of muscle-invasive urothelial bladder cancer was shown to have prognostic and therapy-predictive relevance and thus may help to inform therapeutic decision-making. However, current classification systems rely on whole transcriptome analysis, which is expensive, requires higher amounts of tissue samples, and therefore is not compatible with the daily clinical routine. Therefore, we developed a simple and robust gene panel-based classifier method to reproduce various relevant molecular classification systems (TCGA, MDA, GSC, LundTax, and Consensus). This approach was then tested on institutional cohorts of frozen and formalin-fixed and paraffin-embedded tissue samples using reverse transcription quantitative PCR and NanoString analyses. Here, we provide a step-by-step description of our panel-based subtype classifier method.

A simple and sensitive method for the determination of methylene blue and its analogues in fish muscle using UPLC-MS/MS.

A new, simple and sensitive method for determining and confirming methylene blue and its analogues such as azure A, azure B, azure C, thionine, and new methylene blue in fish muscles have been developed. The method is based on acetonitrile extraction followed by extract purification using dispersive solid-phase extraction (dSPE) with basic aluminium oxide (ALN) and solid-phase extraction (SPE) using primary and secondary amines (PSA) sorbent in matrix adsorption mode. The separation and detection of the dyes in the fish extract are achieved within 5 min by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) using an octadecyl analytical column with a mixture of acetonitrile, methanol and 0.1% formic acid as a mobile phase in gradient elution. The developed method has been in-house validated according to European law. The method recovery for fish muscle was 98.3-103.1%, whereas the decision limit (CCα) was from 0.45 to 0.49 µg kg-1.

TWEAK and Fn14 are overexpressed in immune-mediated necrotizing myopathy: implications for muscle damage and repair.

OBJECTIVES: TNF-like weak inducer of apoptosis (TWEAK) and its sole receptor fibroblast growth factor-inducible 14 (Fn14) are involved in various inflammatory conditions. This study was performed to investigate the potential role of TWEAK/Fn14 in immune-mediated necrotizing myopathy (IMNM). METHODS: Muscle biopsies from patients with IMNM (n = 37) and controls (n = 11) were collected. Human muscle cells were treated with TWEAK in vitro. Muscle biopsies and cultured muscle cells were analysed by immunostaining and quantitative PCR. Serum levels of TWEAK and Fn14 were detected by ELISA. RESULTS: TWEAK and Fn14 were overexpressed in IMNM muscle biopsies. The percentage of Fn14-positive myofibers correlated with disease severity, myonecrosis, regeneration and inflammation infiltrates. Fn14-positive myofibers tended to be surrounded or invaded by CD68+ macrophages. TWEAK treatment had a harmful effect on cultured muscle cells by inducing the production of multiple chemokines and pro-inflammatory cytokines. Serum Fn14 levels were increased in patients with IMNM and correlated with muscle weakness. CONCLUSIONS: TWEAK/Fn14 signalling was activated in IMNM, most likely aggravating muscle damage via amplifying inflammatory response and macrophages chemotaxis. Fn14 seems to be a biomarker for assessing disease severity in IMNM. In addition, Fn14 may also contribute to muscle injury repair.

Meta-analysis and health risk assessment of toxic heavy metals in muscles of commercial fishes in Caspian Sea.

Heavy metals from contaminated seafood can have serious consequences for human health. Several studies on the amount of heavy metals in Caspian Sea fish have been conducted to ensure their food safety. This meta-analysis aimed to investigate the levels of five toxic heavy metals; Lead (Pb), Cadmium (Cd), Mercury (Hg), Chromium (Cr), and Arsenic (As) in muscles of commercial fishes of the Caspian Sea with assessing their health risk of developing oral cancer in terms of fish collection place and fish type. A systematic search was performed, and the random effect model was applied for meta-analysis. Finally, 14 studies with 30 different sets of results were included. Our results showed that the pooled estimates for Pb, Cd, Hg, Cr, and As were 0.65 mg/kg (0.52, 0.79), 0.08 mg/kg (0.07, 0.10), 0.11 mg/kg (0.07, 0.15), 1.77 mg/kg (1.26, 2.27), and 0.10 mg/kg (- 0.06, 0.26) respectively. The levels of Pb and Cd were higher than their (FAO/WHO) maximum limits. The Estimated Daily Intake (EDI) of Pb and Cd in Mazandaran, and Hg in Gilan were more than Total Daily Intake (TDI) limits. The consumers' non-carcinogenic risk (THQ) for Hg in Mazandaran and Gilan, and As in Gilan, was unsafe. Carcinogenic risk (CR) for Cr and Cd in all three provinces, and for As in Mazandaran and Gilan, was greater than 1*10 - 4 and was unsafe. The lowest and highest risk levels of oral cancer were for Rutilus kutum and Cyprinus carpio respectively.

A combined survey and exposure study reveals the distribution characteristic of chromium in fish head.

In this study, the content of chromium (Cr) in three main edible tissues (dorsal muscle, ventral muscle, and head) of 14 different fish species was determined. As expected, Cr level in fish head is generally higher than that in fish muscle. To elucidate the deposition of Cr in the fish head, bighead carp (Hypophthalmichthys nobilis) was chosen as the model fish and a 14-day-exposure experiment of 1 mg/L Cr was subsequently conducted. The fish head was divided into 7 parts, and the results showed that, in different parts of the head, Cr was significantly accumulated in the brain, the muscle after gills, and the muscle of others, and the bone after the fish was exposed to 1-mg/L Cr for 14 days. Additionally, the health risk assessment showed that there was no potential non-carcinogenic risk of consuming the dorsal muscle, the ventral muscle, and the head of the 14 fish species to children, teenagers, and adults as the THQ values were less than 1. However, there was potential carcinogenic risk of consuming the fish head for adults in the 14 fish species. It is worth noting that, in the exposure experiment, the consuming of the lips and the bone of fish head also posed potential carcinogenic risk to adults while there was no potential risk of consuming the total fish head. Overall, compared to the fish muscle, the fish head usually had a high accumulation of Cr, and the Cr was mainly deposited in the position of the bone, which may pose potential carcinogenic risk to adults. Thus, the potential health risk of consuming the fish head is mainly associated with the content of the bone in the head, and, in the health risk assessment, the influence of the bone in the head should be considered, which may overstate the risks as fish head bone is usually not to be eaten.

The pattern of MHC class I expression in muscle biopsies from patients with myositis and other neuromuscular disorders.

OBJECTIVE: Diagnostic muscle biopsies are routinely immunostained for major histocompatibility complex class I (MHC-I) protein. In this study we analysed the prevalence and patterns of MHC-I immunostaining in biopsies from patients with different types of myopathies and neurogenic disorders. METHODS: All 357 diagnostic muscle biopsies processed at the Johns Hopkins Neuromuscular Pathology Laboratory from August 2013 to January 2017 were immunostained for MHC-I. The prevalence and patterns of MHC-I immunostaining were compared between patients with histologically normal muscle biopsies (n = 31), idiopathic inflammatory myopathies (IIMs; n = 170), non-inflammatory myopathies (n = 60) and neurogenic disorders (n = 96). RESULTS: MHC-I immunostaining was abnormal in most patients with DM (98%), sporadic IBM (sIBM; 100%), immune-mediated necrotizing myopathy (IMNM; 100%) and polymyositis (77%). In contrast, MHC-I immunostaining was less frequently present in non-inflammatory myopathies (32%) or neurogenic disorders (30%). Overall, abnormal MHC-I immunostaining had a sensitivity of 0.95 and a specificity of 0.82 for diagnosing IIMs. A focal MHC-I staining pattern was associated with IMNM, whereas a global pattern was more prevalent in sIBM and a perifascicular pattern was significantly more common in dermatomyositis. Among 18 DM biopsies without perifascicular atrophy, 50% had a perifascicular MHC-I staining pattern. Sarcoplasmic upregulation staining was more common than sarcolemmal staining across all groups. CONCLUSION: MHC-I immunostaining was useful to distinguish IIMs from non-inflammatory myopathies or neurogenic disorders. Of note, a perifascicular MHC-I staining pattern was present only in those with DM, including half of those without perifascicular atrophy; many of these biopsies may not otherwise have been diagnostic for DM.

Evaluation of the Effect of Different Cooking Methods on the Heavy Metal Levels in Crayfish Muscle.

The current study investigated the effects of various cooking styles (boiling, frying, and steaming) and seasoning methods (home cooking and ready-to-eat commodity) on levels of nine heavy metals in the crayfish (Procambarus clarkii) muscle. The estimated daily intake (EDI), target hazard quotients (THQ), and target cancer risk (TCR) were used to assess the health risk in the crayfish muscle. The results showed that cooking processes significantly increased the concentration of Cu, which raises a potential risk for children (the THQ values > 1). The levels of toxic heavy metals in the ready-to-eat crayfish muscle were significantly higher than those in household cooking. Especially for As, the THQ values rose to 7.1 and 13.2 for adults and children respectively. Therefore, home cooking is safer than ready-to-eat crayfish, and children should consume crayfish within a limited range. The recommended consumption of the cooked abdominal muscle of crayfish should be 257 and 58 g/day, for children (16 kg) and adults (70 kg), respectively.

Cyhalofop-butyl and pyribenzoxim-induced oxidative stress and transcriptome changes in the muscle of crayfish (Procambarus clarkii).

Cyhalofop-butyl and pyribenzoxim are commonly used herbicides in rice-crayfish co-culture fields. In actual production, weed control in paddy fields is inseparable from cyhalofop-butyl and pyribenzoxim, while its risk to P. clarkii is still unclear. The present study investigated the risk of acute and subchronic toxicity of cyhalofop-butyl and pyribenzoxim to P. clarkii. The results showed that cyhalofop-butyl and pyribenzoxim exposure for 28 days could accumulate in P. clarkii muscle and inhibit P. clarkii growth. Further research found that the malondialdehyde (MDA) level and glutathione-S-transferase (GST) activity in muscle of P. clarkii were significantly increased after exposure to cyhalofop-butyl and pyribenzoxim (4 days and 28 days), and the superoxide dismutase (SOD) and catalase (CAT) activities were significantly altered. Histological results also confirmed cyhalofop-butyl and pyribenzoxim-induced muscle damage in P. clarkii. Additionally, after 28 days exposure to 1.02 mg/L cyhalofop-butyl and 10.4 mg/L pyribenzoxim, transcriptome analysis identified 2029 and 4246 differentially expressed genes (DEGs), respectively. Exposure to 1.02 mg/L cyhalofop-butyl significantly altered metabolism-related pathways, such as drug metabolism-other enzymes, glutathione metabolism, drug metabolism-cytochrome P450, fatty acid biosynthesis and fatty acid degradation. While the pathways related to antioxidant system and nutrient substances synthesis and metabolic were significantly enriched after exposure to 10.4 mg/L pyribenzoxim. This research has significant implications for scientific and rational use of herbicides under rice-crayfish co-culture and will contribute to the development of the highly productive agricultural model.

Health risk assessment of heavy metal concentration in muscle of Chelon auratus and Chelon saliens from the southern Caspian Sea.

Heavy metals are one of the most serious pollutants in aquatic ecosystems, and their accumulation in fish products causes harmful effects on human health. In this context, we set out to determine the concentrations of heavy metals in the muscle of two fish species of commercial interest, Chelon auratus and Chelon saliens on the south coast of the Caspian Sea. We aimed to assess the degree of environment contamination in this area and to estimate the potential risk to human health derived from the consumption of fish. The mean concentrations of the different metals analysed were very varied in both species. In fact, some concentrations exceeded the permissible limits for the protection of human health for Cd and Pb, and some values of As were above those referenced by other authors in same species in the Caspian Sea. None of the estimated daily intake values exceeded the tolerable intake based on the consumption under consideration. Nonetheless, the accumulative hazard values evidenced a potential risk to human health, Pb and Hg being those giving a higher target hazard quotient. The cancer risk from exposure to As from fish consumption in children was above the "acceptable" risk to life. Thus, in view of the accumulative nature of heavy metals, a moderate and non-abusive fish consumption in this area, particularly in children, would be recommendable.

Health risk assessment of heavy metals (Zn, Pb, Cd, and Hg) in water and muscle tissue of farmed carp species in North Iran.

This cross-sectional study was conducted to determine and compare the concentrations of heavy metals (Zn, Pb, Cd, and Hg) in carp-farming water and muscle of various carp species including common carp (Cyprinus carpio), bighead carp (Hypophthalmichthys nobilis), silver carp (Hypophthalmichthys molitrix), and grass carp (Ctenopharyngodon idella) collected from three major warm-water fish farms in Mazandaran Province (Iran) during March 2018 to March 2019. In addition, bioaccumulation of heavy metals (BCFs) and carcinogenic and non-carcinogenic risk assessments of consumers exposed to heavy metals through fish consumption were estimated. The water concentration of all metals in this study was lower than permissible limits. The concentration of Zn in the water (10.21-17.11 µg L-1) was higher than that of other metals in all sites, followed by Pb > Cd > Hg. In fish muscle, Zn concentration in silver carp was the highest, and the lowest concentrations were related to Hg and Cd in common carp and grass carp, respectively. The target hazard quotients (THQ) indicated that the non-carcinogenic health risk to humans was relatively low by consuming four farmed carp species products. The carcinogenic risk of inorganic Pb was 1.24E-04 (common carp) to 2.11E-04 (grass carp) for adults, which is within the acceptable range. The values of BCFs for all metals demonstrated that farmed carp muscle could not be considered a bioaccumulative tissue for heavy metals. The results indicated that the concentrations of heavy metals in the farmed carp species in North Iran were relatively low and did not cause considerable human health risks.

Urine PD-L1 is a tumor tissue candidate substitute and is associated with poor survival in muscle-invasive bladder cancer patients.

Programmed death molecule ligand 1 (PD-L1) expression in urothelial carcinoma is a predictive marker used to guide immunotherapy. As expression of PD-L1 may be heterogeneous in the tumor tissue space, it cannot be accurately determined by immunohistochemical analysis. In this study, we examined PD-L1 protein levels in preoperative urine samples from bladder cancer patients, evaluated the prevalence of PD-L1 in urine, examined the usefulness of urine as a surrogate for PD-L1 expression in tumors, and compared PD-L1 expression in postoperative pathological sections. We found that PD-L1 in urine and tumor tissue correlated well and that it may be able to some extent serve as a surrogate for tissues in bladder cancer and thus predict risk of recurrence in muscle-invasive bladder cancer (MIBC) patients. Our findings reveal the clinical relevance of urine PD-L1 as a noninvasive prognostic indicator for immunotherapy and offer clinical translational suggestions for eventual development of a prognostic model for immunotherapy for bladder cancer.