Liver Specimens Research Articles

Integrative multiomic analysis identifies distinct molecular subtypes of NAFLD in a Chinese population

Nonalcoholic fatty liver disease (NAFLD) has become a common health care burden worldwide. The high heterogeneity of NAFLD remains elusive and impairs outcomes of clinical diagnosis and pharmacotherapy. Several NAFLD classifications have been proposed on the basis of clinical, genetic, alcoholic, or serum metabolic analyses. Yet, accurately predicting the progression of NAFLD to cirrhosis or hepatocellular carcinoma (HCC) in patients remains a challenge. Here, on the basis of a Chinese cohort of patients, we classified NAFLD into three distinct molecular subtypes (NAFLD-mSI, NAFLD-mSII, and NAFLD-mSIII) using integrative multiomics including whole-genome sequencing (WGS), proteomics, phosphoproteomics, lipidomics, and metabolomics across a broad range of liver, blood, and urine specimens. We found that NAFLD-mSI had higher expression of CYP1A2 and CYP3A4, which alleviate hepatic steatosis through mediating free fatty acid/bile acid–mTOR–FXR/PPARα signaling. NAFLD-mSII displayed an elevated risk of liver cirrhosis along with increased hepatic infiltration of M1 and M2 macrophages because of lipid-triggered hepatic CCL2 and CRP production. NAFLD-mSIII exhibited a potential risk for HCC development by increased transcription of CEBPB- and ERCC3-regulated oncogenes because of activation of the EGF-EGFR/CHKA/PI3K-PDK1-AKT cascade. Next, we validated the existence of these three NAFLD molecular subtypes in an external cohort comprising 92 patients with NAFLD across three different Chinese hospitals. These findings may aid in understanding the molecular features underlying NAFLD heterogeneity, thereby facilitating clinical diagnosis and treatment strategies with the aim of preventing the development of liver cirrhosis and HCC.

Discovery of live material of Marginella (Roseamarginella) namaqua

Recently (March 2024) the authors undertook another field trip to the Kalahari and northern West Coast of South Africa. In March 2021 the authors had an expedition through the Kalahari, Karoo and West Coast of South Africa that took them to a little-known fisher’s town, Hondeklip Bay. For background on the town, in 1846, Thomas Grace (a ship’s captain) discovered a small natural harbor and he established a trading station, naming the town after a gneiss rock which vaguely resembles a dog. The name of the town is “Hondeklipbaai”, an Afrikaans name that translates as “dog stone bay” in English. It became the preferred harbor of export for copper ore, until the harbor in Port Nolloth was developed in the 1850s. Alluvial diamond-mining operations started in the area in 1970 and constitutes the main business of mines in that area of the Northern Cape (Mohomed 2006). Just north of the Hondeklip Bay lighthouse a lesser-known bay is found, “Moordenaarsbaai”, where the study material that were used for describing the species was collected. Unfortunately, at that stage the authors could only spend one and a half day in the area, and no live material could be collected due to the neat tide and stormy seas. The recent trip to Hondeklip Bay by the authors did not disappoint, the tide timing was perfect with springtide and good weather. A bit of scratching between the rocks and sea grass in rough black-muddy sand revealed three live specimens. What makes this special is that this species so far only been found in this area and this is the first livematerial.

Cleotychini planthoppers from Perth region in Western Australia: The new genus Femotyche gen. nov., and more… (Hemiptera: Fulgoromorpha: Dictyopharidae)

The new genus Femotyche gen. nov. is described to accommodate two new species from Western Australia: Femotyche kerryae sp. nov. from Perth area and Femotyche hortorum sp. nov. from York. Illustrations of male and female type specimens and a distribution map are provided. The male terminalia as well as the female terminalia, habitat, host plant and live specimens of the new species are illustrated when available. The tribe Cleotychini now contains two genera, the genus Cleotyche Emeljanov, 1997 with two subgenera and five species and the genus Femotyche gen. nov. with two species. Data are provided on an additional, undescribed genus from the same geographic area, recorded from two species known only from photographs.

A self‐cooling self‐humidifying mosquito carrier backpack for transporting live adult mosquitoes on foot over long distances under challenging field conditions

AbstractIt is often necessary to use motorised transport to move live mosquitoes from distant field collection points into a central insectary, so that their behavioural and/or physiological phenotypes can be assessed under carefully controlled conditions. However, a survey of heritable insecticide susceptibility traits among wild‐caught Anopheles arabiensis mosquitoes, collected across an extensive study area composed largely of wilderness in southern Tanzania, necessitated that live mosquitoes were carried on foot over distances up to 25 km per day because most of the area was impassable by car, motorcycle or even bicycle during the rains. A self‐cooling, self‐humidifying carrier backpack was therefore developed that allows live adult mosquito specimens to be transported across rugged miombo woodland and floodplain terrain throughout the year. This wettable backpack was fabricated from stitched Tanzanian kitenge cotton fabric and polyvinyl chloride–coated fibreglass netting that allows easy circulation of air in and out. An outer cover flap made of cotton towelling embedded inside a kitenge envelope overhangs the fibreglass netting upper body of the bag, to protect mosquitoes from direct sunlight, and can be soaked with water to maintain low temperature and high humidity inside. Mean survival of insectary‐reared female An. arabiensis transported through nine different mobile camps inside the 509 km2 Ifakara‐Lupiro‐Mang'ula wildlife management area (ILUMA WMA), over up to 143 km and 25 days, was statistically indistinguishable from those left in the field insectary over the same period. Although considerable variance of survival was observed between different batches of mosquitoes from the insectary and between individual cups of mosquitoes, the different levels and positions inside the backpack had no influence on this outcome. Temperature and humidity inside the backpack were maintained at standard insectary conditions throughout, despite much more extreme conditions immediately outside. When the backpack was used to transport wild An. arabiensis and Anopheles quadriannulatus across a much larger study area of >4000 km2, encompassing the ILUMA WMA, some nearby villages and adjacent parts of Nyerere National Park (NNP), it achieved a mean survival rate of 58.2% (95% confidence interval 47.5–68.2). Encouragingly, no difference in survival was observed between ILUMA WMA and NNP even though transport back from NNP involves much longer distances, sometimes involving lengthy journeys by car or boat. Overall, this mosquito carrier backpack prototype appears to represent a viable and effective method for transporting live wild‐caught mosquitoes on foot across otherwise impassable terrain under challenging weather conditions with minimal detrimental impact on their survival.

High PD-1 and CTLA-4 expression correlates with host immune suppression in patients and a mouse model infected with Echinococcus multilocularis

BackgroundAlveolar echinococcosis (AE), a fatal disease caused by Echinococcus multilocularis, often affects the liver, with tumor-like growth. However, the mechanism by which E. multilocularis evades host immune surveillance remains unclear.MethodsWe collected liver specimens from hepatic alveolar echinococcosis (HAE) patients and established a mouse model of E. multilocularis infection. Immunofluorescence staining and flow cytometry were performed to analyze programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte associated antigen 4 (CTLA-4) expression in human samples, while flow cytometry and quantitative real-time polymerase chain reaction (PCR) were performed for similar analyses in mouse samples. Cell proliferation and protoscolex (PSC) killing assays were designed to explore how E. multilocularis induces host immunosuppression.ResultsAn inflammatory reaction band with high PD-1 and CTLA-4 expression was found in close liver tissue (CLT). The ratio of regulatory T cells (Tregs) was higher in CLT than in distant liver tissue (DLT), and Tregs in CLT tended to express higher levels of PD-1 and CTLA-4 than those in DLT from HAE patients. Echinococcus multilocularis-infected mice showed significantly elevated expression of PD-1 and CTLA-4 on splenocytes and peritoneal cells. PD-1/PD-L1 or CTLA-4 pathway blockade could relieve the immunosuppressive effects of Tregs from infected mice and enhance PSC killing by mouse splenocytes.ConclusionsE. multilocularis regulated the function of T cells via the PD-1/PD-L1- and CTLA-4-dependent pathways and subsequently evaded host immune attacks. These findings provide insights for investigating the pathogenic mechanism of AE.Graphical

Features of liver structural organization in posttraumatic stress disorder

Posttraumatic stress disorder (PTSD) is a severe psychological condition resulting from traumatic experiences, leading to long-term physiological changes in the body. It affects mental health and induces hormonal imbalances that disrupt the hypothalamus-pituitary-adrenal axis, influencing multiple organ systems. While PTSD is often linked with comorbid conditions like depression and anxiety, its effects on the hepatobiliary system are less understood. Aim. This study aimed to assess PTSD's impact on liver morphology in rats of different sexes and compare the severity of changes between males and females, contributing to the understanding of stress-related physiological effects. Materials and Methods. The experiment involved 40 Wistar rats, divided into control and PTSD-induced groups for both sexes. A validated stress induction protocol was used, followed by a thorough morphological analysis of liver specimens fixed in 10% formalin and stained with hematoxylin-eosin. Histopathological evaluation focused on liver architecture, hepatocyte condition, vascular integrity, and cellular organization. Results. Control animals displayed standard liver structure, while PTSD-induced groups exhibited significant morphological changes, such as dystrophic alterations in hepatocytes, congestion in hepatic veins, and the formation of blood clots. Female rats showed more pronounced changes than males, including hepatocyte swelling and vacuolization, indicating heightened cellular stress and reduced regenerative capacity. Conclusions. The findings highlight that PTSD-related liver changes were more significant in female rats, emphasizing the importance of considering gender differences in stress-related disorders. These alterations suggest a complex interaction between chronic psychological stress and liver physiology, warranting further research on mechanisms by which PTSD affects the hepatobiliary system and potential therapeutic interventions. for at-risk populations.

EEF1A2 identified as a hub gene associated with the severity of metabolic dysfunction-associated steatotic liver disease.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent chronic liver disease that ranges from metabolic dysfunction-associated steatotic liver (MASL) to metabolic dysfunction-associated steatohepatitis (MASH), and may eventually progress to cirrhosis and hepatocellular carcinoma (HCC). The underlying mechanism of MASLD remains incompletely understood. This study aimed to identify key gene implicated in MASLD pathogenesis and validate its correlation with disease severity through an integration of bioinformatics and experimental approaches. Liver transcriptome data from MASLD patients were obtained from the Gene Expression Omnibus (GEO) database. A diet-induced MASLD mouse model was developed, and liver RNA-sequencing was performed. Liver specimens and clinical data from patients were collected for further analysis. A total of 120 differentially expressed genes (DEGs) were shared between datasets GSE89632 and GSE213621, with functional enrichment in inflammatory, metabolic, and cell cycle-related pathways. Protein-protein interaction (PPI) network analysis identified three modules associated with MASLD, with the cell cycle-related module being the most notable. EEF1A2 was identified as a novel hub gene and revealed to be elevated with MASLD progression through dataset analysis. EEF1A2 was confirmed to be highly expressed in the livers of both MASLD mouse models and patients. Moreover, the increased expression of EEF1A2 in MASH was positively correlated with higher serum alanine aminotransferase (ALT), alanine aminotransferase (AST), total cholesterol (TC), and body mass index (BMI). In conclusion, EEF1A2 is a novel hub gene significantly associated with MASLD severity and is a promising biomarker and therapeutic target for MASLD.

A conspectus of the genus Pigea (Violaceae) in India

An updated conspectus of the genus Pigea in India is provided based on live and herbarium specimens. We recognize three species, namely P. enneasperma, P. indica and P. travancorica and exclude three species from the National Flora, namely P. vatsavayae ( Hybanthus vatsavayae), P. stellarioides ( H.stellarioides) and P. puberula ( H. puberulus). Detailed notes are provided.

Caracterización morfológica de pelos de guardia de seis especies mamíferos pequeños no voladores del occidente de Ecuador

This study presents the morphological characterization of guard hairs from six species of small non-flying mammals. Guard hair samples were obtained from both live individuals and museum specimens, with cuticular and medullary impressions made from the middle portions of the shafts and shields, respectively. A total of 112 samples were collected from the following species: Marmosa simonsi, Metachirus myosorus, Sigmodon peruanus, Aegialomys xanthaeolus, Rattus sp., and Mus musculus. This information can be used for taxonomic identification and, for instance, in the identification of prey items in carnivore and nocturnal raptor diet studies, which are areas lacking information in western Ecuador.

Two new records of sea anemones (Cnidaria, Anthozoa, Actiniaria) from the Eastern Pacific coast of El Salvador

Two species of the genus Actinostella Duchassaing, 1850 are documented for the first time on the coasts of El Salvador: A. bradleyi (Verrill, 1869) and A. californica (McMurrich, 1893). Both species are briefly described, and images of live specimens showing their main external features are provided, as well as images of histological sections showing some taxonomic characteristics of their internal anatomy. These two species are distinguished from each other primarily by the number of tentacles, the number of mesentery pairs, the arrangement of gametogenic tissue, and some differences in their cnidom. The record of these two species in El Salvador increases the number of sea anemones recorded for the country to 12. Additionally, an updated list of sea anemone species documented in El Salvador is provided.

Evaluation of Minimum-to-Severe Global and Macrovesicular Steatosis in Human Liver Specimens: A Portable Ambient Light-Compatible Spectroscopic Probe.

Hepatic steatosis (HS), particularly macrovesicular steatosis (MaS), influences transplant outcomes. Accurate assessment of MaS is crucial for graft selection. While traditional assessment methods have limitations, non-invasive spectroscopic techniques like Raman and reflectance spectroscopy offer promise. This study aimed to evaluate the efficacy of a portable ambient light-compatible spectroscopic system in assessing global HS and MaS in human liver specimens. A two-stage approach was employed on thawed snap-frozen human liver specimens under ambient room light: biochemical validation involving a comparison of fat content from Raman and reflectance intensities with triglyceride (TG) quantifications and histopathological validation, contrasting Raman-derived fat content with evaluations by an expert pathologist and a "Positive Pixel Count" algorithm. Raman and reflectance intensities were combined to discern significant (≥ 10%) discrepancies in global HS and MaS. The initial set of 16 specimens showed a positive correlation between Raman and reflectance-derived fat content and TG quantifications. The Raman system effectively differentiated minimum-to-severe global and macrovesicular steatosis in the subsequent 66 specimens. A dual-variable prediction algorithm was developed, effectively classifying significant discrepancies (> 10%) between algorithm-estimated global HS and pathologist-estimated MaS. Our study established the viability and reliability of a portable spectroscopic system for non-invasive HS and MaS assessment in human liver specimens. The compatibility with ambient light conditions and the ability to address limitations of previous methods marks a significant advancement in this field. By offering promising differentiation between global HS and MaS, our system introduces an innovative approach to real-time and quantitative donor HS assessments. The proposed method holds the promise of refining donor liver assessment during liver recovery and ultimately enhancingtransplantation outcomes.

Are the postmortem concentration changes of the synthetic cannabinoid cumyl-5F-P7AICA and its N-pentanoic acid metabolite dependent on the environmental conditions? – A systematic study following pulmonary administration to pigs

The number of fatal cases involving synthetic cannabinoids (SCs) is increasing. However, interpretation of postmortem (PM) toxicological findings is challenging, due to unknown PM intervals and possible redistribution processes or instabilities. Only sparse data on SCs are available. Therefore, a controlled pig study was performed to determine the PM stability of cumyl-5F-P7AICA under different environmental conditions. Ten pigs inhalatively received 200 µg/kg body weight cumyl-5F-P7AICA each. Six hours later, they were euthanized and biopsies of the main tissues and body fluids were taken. Animals were stored in air or water (n=5 each) and samples were repeatedly taken for three days. Quantification of cumyl-5F-P7AICA and its N-pentanoic acid metabolite (NPA) was performed using standard addition or a fully validated method (blood) followed by LC-MS/MS. Time-dependent concentration changes of cumyl-5F-P7AICA were observed in liver, bile fluid and muscle specimens at both conditions. Concentrations of NPA only changed considerably in duodenum content of animals stored in air. Environment-dependent concentrations changes were only observed for cumyl-5F-P7AICA in kidney and the NPA metabolite in duodenum content. Overall, cumyl-5F-P7AICA and its metabolite seem to be quite stable in PM specimens. Hence, also central blood might be analyzed, if no peripheral blood is available.

Characterizing alcohol-related and metabolic dysfunction-associated steatotic liver disease cirrhosis via fibrotic pattern analysis

This study aimed to address the diagnostic challenges in distinguishing between alcohol-related liver disease (ALD) and metabolic dysfunction-associated steatotic liver disease (MASLD). We utilized whole-slide imaging technology to conduct a comprehensive digital analysis of liver specimens collected from patients undergoing transplantation. This study included 36 and 17 patients with ALD and MASLD cirrhosis, respectively, who underwent transplantation at our institution. Digital slides were analyzed for fibrosis patterns using FibroNest™. Patient background characteristics were comparable between ALD (n = 36) and MASLD (n = 17) groups, except for sex. The ALD group exhibited thicker collagen per strand, longer and more flexural fibrosis, and a more heterogeneous distribution than the MASLD group. In patients with ALD and concomitant metabolic dysfunction, fiber distribution became relatively uniform, resembling MASLD. Application of the phenotypic fibrosis composite score achieved 100% sensitivity and specificity for ALD/MASLD diagnosis. Digital pathological analysis of the fibrosis patterns showed morphological differences between ALD and MASLD. This approach holds promise for histological differentiation, providing valuable insights beyond the current definitions based solely on alcohol intake. This study emphasizes the potential of digital pathology in refining the diagnostic criteria for hepatic disorders.

Reproductive ecology and egg parasitism of the Samoan swallowtail butterfly

AbstractWe investigated the reproductive ecology and effects of egg parasitism on the Samoan swallowtail butterfly (Papilio godeffroyi), which survives only on Tutuila Island, American Samoa, after having disappeared from the much larger islands of Upolu and Savai‘i in independent Samoa. During monthly surveys of its only known host plant, Micromelum minutum, across eight sites in 2013 and 2014, we collected eggs, eggshells, larvae, pupae, and pupal exuviae. Live specimens were reared under laboratory conditions to determine reproductive outcomes, developmental rates, and sex ratios, as well as parasitoid attack frequencies, brood sizes, and sex ratios. Sixty‐six of 448 (14.7%) eggs produced larvae, 47 of which became adults. The sex ratio was approximately even overall and within each developmental stage. Eggs were slightly larger on individual host trees and in host tree stands that yielded more eggs per unit of foliage, indicating that ovipositing females responded to some features of host trees and stands. Eggs hatching female or male larvae were similar in size, and the sexes developed at similar rates. A newly described species of parasitoid wasp, Ooencyrtus pitosina (Encyrtidae), emerged from 73.6% of 382 butterfly eggs that failed to hatch in the laboratory (62.7% of 448 eggs overall). Forty‐one other eggs contained dead parasitoid larvae. An additional, unidentified Ooencyrtus wasp species emerged from a single P. godeffroyi egg. No parasitoids were reared from P. godeffroyi larvae or pupae. Of 656 P. godeffroyi eggshells collected in the field and examined in the laboratory, 62.2% showed signs of having been parasitized by O. pitosina. There was no evidence that parasitism rates were density‐dependent. O. pitosina brood sizes ranged from 1 to 5, with the sex ratio skewed toward females (2.40 F:1.00 M). Larger parasitoid broods were associated with slightly larger host eggs, indicating that female wasps may adjust brood size according to host egg size or that fewer wasp larvae are able to complete development in smaller eggs. Techniques used to rear both P. godeffroyi and O. pitosina in the laboratory could be applied to a captive‐rear, wild‐release program, which may facilitate reestablishment of the species in Samoa.

Outcomes of CEM43 in Predicting Thermal Damage Induced by Focal Laser Ablation in Controlled Ex Vivo Experiments: A Comparison to Histology and MRI.

Focal laser ablation (FLA) serves as a targeted therapy for prostate cancer (PCa). Clinical studies have demonstrated significant variations in ablation volumes with consistent fiber configurations. Consequently, a prediction model is needed for the safe application of FLA in treating PCa. This study aimed to evaluate the reproducibility of FLA-induced temperature profiles in controlled ex vivo experiments using clinical laser treatment protocols. Additionally, it sought to examine the effectiveness of the CEM43 model in predicting the zone of irreversible damage (ZID) and to compare these findings with outcomes derived from the Arrhenius model. Freshly excised postmortem human prostate and porcine liver specimens were used for controlled ex vivo ablation. Tissues were secured in a Perspex sample holder for precise placement of the laser fiber and thermocouples. FLA was conducted with a 1064-nm Nd:YAG laser at 3 W in continuous-wave mode for 10 min. Pre- and post-FLA 3D T1-weighted 7 T MRI scans were obtained to assess the treatment area. Whole-mount hematoxylin and eosin histological slides were prepared and digitized. On histology, the ZID was defined as the total of vaporized, carbonized, and coagulated tissue. A 2D thermal development map was created from temperature data, using bi-cubic interpolation. The cumulative equivalent thermal isoeffect dose at 43°C in minutes (CEM43) model was applied to predict the ZID, with 240 equivalent minutes (240-CEM43) used as the damage threshold. Additionally, the Arrhenius thermal model was used for comparison of CEM43 results. Predicted ZIDs were compared to MRI and histology. FLA treatment was performed on ex vivo human prostate samples (n = 2) and porcine liver specimens (n = 5). For human prostate tissue, FLA did not result in an identifiable ZID upon histological macroscopic examination or a lesion on MRI. Ex vivo porcine liver samples showed a clearly demarcated oval-shaped hyperintense lesion surrounding the laser fiber tip on post-FLA MRI. The MRI lesion (range 1.6-2.1 cm2) corresponded with the shape and location of the ZID on histology, but was smaller (median 1.7 vs. 3.2, p = 0.02). Histological examination of porcine liver samples revealed ZIDs ranging from 2.1 to 4.1 cm2, whereas 240-CEM43-predicted ZIDs ranged from 3.3 to 3.8 cm2. Although the median 240-CEM43-predicted ZID was not significantly larger than the histology ZID (3.8 vs. 3.2 cm2, p = 0.22), it tended to overpredict the histological results in most experiments. The median Arrhenius-predicted ZID was similar to the histologicalZID (3.2 vs. 3.2 cm2, p = 0.56), but varied in size when comparing individual experiments (range 2.5-3.2 cm2). FLA on ex vivo human prostate showed no thermal damage on histopathology or MRI. Ex vivo porcine liver FLA resulted in identifiable ZID on histology and lesions on MRI. 240-CEM43 generally overestimated the ZID and had less variability compared to histology. Results from the Arrhenius model were in better agreement with the histology findings, but still did not predict the individual FLA-induced histological thermal damage. Inter-experiment ZID variability underlines the need for developing a more comprehensive predictive dosimetry model for FLA in PCa treatment.

The hepatoprotective effect of sodium butyrate on hepatic inflammatory injury mediated by the NLRP3 inflammatory pathway in subchronic fluoride-exposed mice.

Excessive subchronic fluoride exposure can cause severe damage to detoxification organs, including the liver. Sodium butyrate has anti-inflammatory, antitumor, antioxidant and immunomodulatory properties. However, relatively few studies have investigated the effects of sodium butyrate on liver injury caused by subchronic fluoride exposure. The purpose of this research was to investigate the effect and mechanism of sodium butyrate on fluoride-induced hepatic inflammatory injury via the expression of nod-like receptor protein 3 (NLRP3). Mice were subjected to randomization into four groups, control group (C), fluorosis group (F), sodium butyrate alone group (S), and treatment group (Y). The mice in groups F and F + S drank 100mg/L sodium fluoride-containing distilled water freely every day. After fluoride exposure lasted for 3 months, the mice in group S and F + S were gavaged with sodium butyrate daily at a concentration of 1000mg/kg. Following the treatment regimen, liver specimens were collected for analysis. The mRNA and protein expression levels of inflammatory factors and NLRP3 and its downstream gene were measured by RT-qPCR and western blotting. The histological hematoxylin and eosin (H&E) staining of liver showed that the subchronic fluoride-exposed group were chronic inflammation. The liver of treatment group were less vacuolar degeneration and inflammatory infiltration. The results of the biochemical assay showed that the subchronic fluoride-exposed group were liver injury. In addition, the detection of oxidative stress indicators showed that chronic subchronic fluoride exposure could lead to an increase in the level of oxidative stress in the liver, and the treatment alleviated this increase. RT-qPCR results showed that compared with those in the control group, the mRNA levels of the inflammatory factors TNF-α, IL-6 and IL-1β, the NLRP3 inflammasome and its downstream factors NLRP3, caspase-1, gasdermin D (GSDMD) and IL-18 increased in the liver tissue of mice in the subchronic fluoride-exposed group. Sodium butyrate released inflammatory factors during subchronic fluoride exposure and inhibited the protein expression of activated NLRP3 to a certain extent. Sodium butyrate may play a protective role by antagonizing the production of activated inflammasomes and their downstream inflammatory factors in the livers of subchronic fluoride-exposed mice.

Four new species of the lanternfly genus Zanna Kirkaldy, 1902 from Cambodia and Vietnam (Hemiptera: Fulgoromorpha: Fulgoridae)

Four new species of Zanna Kirkaldy, 1902 are described: two from Cambodia, Z. chartieri Constant sp. nov. from Tatai in Koh Kong Province and Z. limbourgi Constant sp. nov. from Phnom Aural Wildlife Sanctuary in Kampong Speu Province and Kbal Spean in Siem Reap Province, and two from Vietnam: Z. bidoupana sp. nov. from Bidoup-Nui Ba National Park in Lam Dong Province and Z. kusamae sp. nov. from Dong Nai Biosphere Reserve in Dong Nai Province. Illustrations of the holotypes and male genitalia, photographs of live specimens and nymphs, a distribution map and host plants records are provided. The type of Zanna chinensis (Distant, 1893) is also illustrated for comparison. The genus Zanna now contains 37 species.

A new species of alien land flatworm in the Southern United States.

Specimens of a flat and dark brown land planarian were found in a plant nursery in North Carolina, USA in 2020. On the basis of examination of photographs of the live specimens only, the specimens were considered as belonging to Obama nungara, a species originally from South America, which has now invaded a large part of Europe. Unexpectedly, a molecular analysis revealed that the specimens did not belong to this species, neither to the genus Obama. We then undertook its histological study, which finally confirmed that the species is a member of the genus Amaga: the species is herein described as a new species, Amaga pseudobama n. sp. The species has been found in three locations in North Carolina and some infested plants were from Georgia. We reinvestigated specimens collected in Florida in 2015 and found that they also belong to this species. Citizen science observations suggest its presence in other states. Therefore, it is likely that A. pseudobama has already invaded a part of south-east USA and that the invasion took place more than ten years ago. The complete 14,909 bp long mitochondrial genome was obtained. The mitogenome is colinear with those of other Geoplanidae and it was possible to find and annotate a tRNA-Thr, which has been reported missing in several geoplanids. Amaga pseudobama shares with other Geoplaninae the presence of alternative start codons in three protein-coding genes of its mitogenome. The availability of this new genome helped us to improve our annotations of the ND3 gene, for which an ATT start codon is now suggested. Also, the sequence of the ATP6 gene raised questions concerning the use of genetic code 9 to translate the protein-coding genes of Geoplanidae, as the whole translated protein would not contain a single methionine residue when using this code. Two maximum likelihood phylogenies were obtained from genomic data. The first one was based on concatenated alignments of the partial 28S, Elongation Factor 1-alpha (EF1) and cox1 genes. The second was obtained from a concatenated alignment of the mitochondrial proteins. Both strictly discriminate A. pseudobama from O. nungara and instead associate it with Amaga expatria. We note that the nine species currently accepted within Amaga can be separated into two groups, one with extrabulbar prostatic apparatus, including the type species A. amagensis, and one with intrabulbar prostatic apparatus, including the new species A. pseudobama. This suggests that species of the latter group should be separated from Amaga and constitute a new genus. This finding again illustrates the possible emergence of new invasive species in regions naturally devoid of large land planarians, such as North America. Amaga pseudobama thus deserves to be monitored in the USA, although its superficial resemblance to O. nungara and Geoplana arkalabamensis will complicate the use of photographs obtained from citizen science. Our molecular information provides tools for this monitoring.

BRUCE liver-deficiency potentiates MASLD/MASH in PTEN liver-deficient background by impairment of mitochondrial metabolism in hepatocytes and activation of STAT3 signaling in hepatic stellate cells.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is currently the most common liver disease, affecting up to 25% of people worldwide, featuring excessive fat accumulation in hepatocytes. Its advanced form, metabolic dysfunction-associated steatohepatitis (MASH), is a serious disease with hepatic inflammation and fibrosis, increasing the need for liver transplants. However, the pathogenic mechanism of MASLD and MASH is not fully understood. We reported that BRUCE ( BIRC6) is a liver cancer suppressor and is downregulated in MASLD/MASH patient liver specimens, though the functional role of BRUCE in MASLD/MASH remains to be elucidated. To this end, we generated liver-specific double KO (DKO) mice of BRUCE and PTEN, a major tumor suppressor and MASLD/MASH suppressor. By comparing liver histopathology among 2-3-month-old mice, there were no signs of MASLD or MASH in BRUCE liver-KO mice and only onset of steatosis in PTEN liver-KO mice. Interestingly, DKO mice had developed robust hepatic steatosis with inflammation and fibrosis. Further analysis of mitochondrial function with primary hepatocytes found moderate reduction of mitochondrial respiration, ATP production and fatty acid oxidation in BRUCE KO and the greatest reduction in DKO hepatocytes. Moreover, aberrant activation of pro-fibrotic STAT3 signaling was found in hepatic stellate cells (HSCs) in DKO mice which was prevented by administered STAT3-specific inhibitor (TTI-101). Collectively, the data demonstrates by maintaining mitochondrial metabolism BRUCE works in concert with PTEN to suppress the pro-fibrogenic STAT3 activation in HSCs and consequentially prevent MASLD/MASH. The findings highlight BRUCE being a new co-suppressor of MASLD/MASH.

Putative role of 6-chogaol against tramadol-induced hepatotoxicity in albino rats via anti-inflammatory, antifibrotic, and antiapoptotic effects

Tramadol is a commonly used drug to relieve pain and avoid premature ejaculation in males with hepatotoxic effects, and 6-chogaol has potent anti-inflammatory and hepatoprotective properties.The work impetus is probing the hepatoprotective mechanisms of 6-chogaol against tramadol hepatoxicity. Twenty adult male rats were enrolled to obtain four equal groups [control group (G1), 6-chogaol group (G2), tramadol group (G3), and 6-chogaol+tramadol group (G4)]. Liver specimens were excised and processed to evaluate hepatocyte injury through histopathological (HP), immunohistochemical (IHC), flow cytometry, and biochemical investigations. The HP study exhibited hepatic injury in G3 hepatocytes (inflammatory cell infiltration, hepatic fibrosis, and disturbed liver structure). The IHC study showed a significant rise in caspase-3 and reduced PCNA immuno-expression (IE). Likewise, the flow cytometry and biochemical experiments exhibited a substantial elevation of apoptotic hepatocytes and the serum levels of IL-1β, IL-6, TNF-α, ALP, ALT, and AST in G3. In contrast, G4 rats significantly improved in all HP, IHC, flow cytometry, and biochemical parameters. Collectively, tramadol intake exerted harmful toxic effects on hepatocytes, whereas 6-Shogaol hampered these changes and served as a natural hepatoprotective agent. Therefore, we advise concurrent intake of 6-Shogaol supplement with tramadol to preserve the integrity of hepatic tissues.