Histological Observations Research Articles

Sugar import mediated by sugar transporters and cell wall invertases for seed development in Camellia oleifera

Abstract Seed development and yield depend on the transport and supply of sugar. However, an insufficient supply of nutrients from maternal tissues to embryos results in seed abortion and yield reduction in Camellia oleifera. In this study, we systematically examined the route and regulatory mechanisms of sugar import into developing C. oleifera seeds using a combination of histological observations, transcriptome profiling and functional analysis. Labelling with the tracer carboxyfluorescein revealed a symplasmic route in the integument and an apoplasmic route for post-phloem transport at the maternal-filial interface. Enzymatic activity and histological observation showed that at early stages (180-220 DAP) of embryo differentiation, the high hexose/sucrose ratio was primarily mediated by acid invertases and the micropylar endosperm/suspensor provides a channel for sugar import. Through Camellia genomic profiling, we identified three plasma membrane-localized proteins including CoSWEET1b, CoSWEET15 and CoSUT2 and one tonoplast-localized protein CoSWEET2a in seeds and verified their ability to transport various sugars via transformation in yeast mutants and calli. In situ hybridization and profiling of glycometabolism-related enzymes further demonstrated that CoSWEET15 functions as a micropylar endosperm-specific gene, together with the cell wall acid invertase CoCWIN9, to support early embryo development, while CoSWEET1b, CoSWEET2a and CoSUT2 function at transfer cells and chalazal nucellus coupled with CoCWIN9 and CoCWIN11 responsible for sugar entry in bulk into the filial tissue. Collectively, our findings provide the first comprehensive evidence of the molecular regulation of sugar import into and within C. oleifera seeds and provide a new target for manipulating seed development.

Apelin receptor modulation mitigates letrozole-induced polycystic ovarian pathogenesis in mice

AimsPolycystic ovarian syndrome (PCOS) is one of the most common (about 5–20%) reproductive disorders in women of reproductive age; it is characterized by polycystic ovaries, hyperandrogenism, and oligo/ anovulation. The levels and expression of ovarian adipokines are deregulated in the PCOS. Apelin is an adipokine that acts through its receptor (APJ) and is known to express in the various tissues including the ovary. It has also been suggested that apelin and APJ could be targeted as therapeutic adjuncts for the management of PCOS. However, no study has been conducted on the management of PCOS by targeting the apelin system. Thus, we aimed to evaluate its impact on combating PCOS-associated ovarian pathogenesis. MethodsThe current work employed a letrozole-induced-hyperandrogenism PCOS-like mice model to investigate the effects of apelin13 and APJ, antagonist ML221. The PCOS model was induced by oral administration of letrozole (1 mg/kg) for 21 days. A total of four experimental groups were made, control, PCOS control, PCOS + aplein13, and PCOS + ML221. The treatment of apelin13 and ML221 was given from day 22 for two weeks.Key findings: The letrozole-induced PCOS-like features such as hyperandrogenism, cystic follicle, decreased corpus luteum, elevated levels of LH/FSH ratio, and up-regulation of ovarian AR expression were ameliorated by apelin13 and ML221 treatment. However, the PCOS-augmented oxidative stress and apoptosis were suppressed by apelin 13 treatments only. ML221 treatment still showed elevated oxidative stress and stimulated apoptosis as reflected by decreased antioxidant enzymes and increased active caspase3 and Bax expression. The expression of ERs was elevated in all groups except control. Furthermore, the PCOS model showed elevated expression of APJ and apelin13 treatment down-regulated its own receptor. Overall, observing the ovarian histology, corpus luteum formation, and decreased androgen levels by both apelin13 and ML221 showed ameliorative effects on the cystic ovary. SignificanceDespite the similar morphological observation of ovarian histology, apelin13 and ML221 exhibited opposite effects on oxidative stress and apoptosis. Therefore, apelin13 (which down-regulates APJ) and ML221 (an APJ antagonist) may have suppressed APJ signalling, which would account for our findings on the mitigation of polycystic ovarian syndrome. In conclusion, both apelin13 and ML221 mediated mitigation have different mechanisms, which need further investigation.

Gene structure, expression and function analysis of the MyoD gene in the Pacific white shrimp Litopenaeus vannamei

The Pacific white shrimp Litopenaeus vannamei is a representative species of decapod crustacean and an economically important marine aquaculture species worldwide. However, research on the genes involved in muscle growth and development in shrimp is still lacking. MyoD is recognized as a crucial regulator of myogenesis and plays an essential role in muscle growth and differentiation in various animals. Nonetheless, little information is available concerning the function of this gene among crustaceans. In this study, we identified a sequence of the MyoD gene (LvMyoD) with a conserved bHLH domain in the L. vannamei genome. Phylogenetic analysis revealed that both the overall protein sequence and specific functional sites of LvMyoD are highly conserved with those of other crustacean species and that they are evolutionarily closely related to vertebrate MyoD and Myf5. LvMyoD expression is initially high during early muscle development in shrimp and gradually decreases after 40 days post-larval development. In adults, the muscle-specific expression of LvMyoD was confirmed through RT–qPCR analysis. Knockdown of LvMyoD inhibited the growth of the shrimp in body length and weight. Histological observation and transcriptome sequencing of muscle samples after RNA interference (RNAi) revealed nuclear agglutination and looseness in muscle fibers. Additionally, we observed significant effects on the expression of genes involved in heat shock proteins, myosins, actins, protein synthesis, and glucose metabolism. These findings suggest that LvMyoD plays a critical role in regulating muscle protein synthesis and muscle cell differentiation. Overall, this study highlights the involvement of LvMyoD in myogenesis and muscle growth, suggesting that it is a potentially important regulatory target for shrimp breeding efforts.

The ever-changing microenvironment of Staphylococcus aureus in cutaneous infections.

Staphylococcus aureus is responsible for the majority of skin and soft tissue infections, which are often diagnosed at a late stage, thereby impacting treatment efficacy. Our study was designed to reveal the physiological changes at different stages of infection by S. aureus through the combined analysis of variations in the skin microenvironment, providing insights for the diagnosis and treatment of S. aureus infections. We established a murine model of skin and soft tissue infection with S. aureus as the infectious agent to investigate the differences in the microenvironment at different stages of infection. By combining analysis of the host immune status and histological observations, we elucidate the progression of S. aureus infection in mice. The results indicate that the infection process in mice can be divided into at least two stages: early infection (1-3 days post-infection) and late infection (5-7 days post-infection). During the early stage of infection, notable symptoms such as erythema and abundant exudate at the infection site were observed. Histological examination revealed infiltration of numerous neutrophils and bacterial clusters, accompanied by elevated levels of cytokines (IL-6, IL-10). There was a decrease in microbial alpha diversity within the microenvironment (Shannon, Faith's PD, Chao1, Observed species, Simpson, Pielou's E). In contrast, during the late stage of infection, a reduction or even absence of exudate was observed at the infected site, accompanied by the formation of scabs. Additionally, there was evidence of fibroblast proliferation and neovascularization. The levels of cytokines and microbial composition gradually returned to a healthy state. This study reveals synchrony between microbial composition and histological/immunological changes during S. aureus-induced SSTIs.

Graft cell expansion from hiPSC-RPE strip after transplantation in primate eyes with or without RPE damage

Clinical studies using suspensions or sheets of human pluripotent cell-derived retinal pigment epithelial cells (hiPSC-RPE) have been conducted globally for diseases such as age-related macular degeneration. Despite being minimally invasive, cell suspension transplantation faces challenges in targeted cell delivery and frequent cell leakage. Conversely, although the RPE sheet ensures targeted delivery with correct cell polarity, it requires invasive surgery, and graft preparation is time-consuming. We previously reported hiPSC-RPE strips as a form of quick cell aggregate that allows for reliable cell delivery to the target area with minimal invasiveness. In this study, we used a microsecond pulse laser to create a local RPE ablation model in cynomolgus monkey eyes. The hiPSC-RPE strips were transplanted into the RPE-ablated and intact sites. The hiPSC-RPE strip stably survived in all transplanted monkey eyes. The expansion area of the RPE from the engrafted strip was larger at the RPE injury site than at the intact site with no tumorigenic growth. Histological observation showed a monolayer expansion of the transplanted RPE cells with the expression of MERTK apically and collagen type 4 basally. The hiPSC-RPE strip is considered a beneficial transplantation option for RPE cell therapy.

The effects of Anchusa azurea methanolic extract on burn wound healing: Histological, antioxidant, and anti-inflammatory evaluation

Anchusa azurea one of the medicinal plants that has been traditionally used for treat burn wounds. However, the traditional claim that A.azurea can hasten burn wound healing has not been supported by scientific studies. This experiment used a male Wistar rats model to investigate the activity of A. azurea aerial parts methanolic extract in burn wound healing. To determine their ability to help in healing burn wounds in rat models, the active components of the aerial parts of A. azurea were extracted with 80% methanol, then, 1% and 10% ointments were prepared from the extract, and applied topically. The LCMS chromatography of A. azurea plant extract showed different active ingredients, including phenolic compounds, flavonoids, fatty acids, and others. The plant extract's investigated as anti-inflammatory, antioxidant, and histological effects on the burn wound healing process. The results showed a significant (p-value < 0.025) rate of burn wound healing with 78.6% and 84.8% contraction, respectively using 1% and 10% (w/w) extract ointments after 12 days. These results were corroborated by histological observations such as collagen deposition, re-epithelialization, and repair of the remaining skin tissues without any sign of cutaneous toxicity. The plant extract showed significant (p-value < 0.025) antioxidant effect at the highest tested dose of 500 µg/mL, scavenging 89.78% of the DPPH with an IC50 of 213.6 µg/mL. These results confirmed by histological changes observations of collagen deposition, re-epithelialization, and reformation of remaining skin tissues without any signs of dermal toxicity. The plant extract exhibited significant (p-value < 0.025) level of antioxidant agents, by scavenging 89.78% of the DPPH at 500 µg/mL with IC50 of 213.6 µg/mL. Additionally, all pro-inflammatory cytokines examined, including IL-6 and IL-10, the results exhibited reduction in IL-6 level and increase IL-10 level. The aerial extract of the A. azurea plant revealed a wealth of several significant active ingredients, including phenolic compounds, flavonoids, fatty acids, and others, suggesting the potential for anti-inflammatory, burn wound-healing, and antioxidant medications. These findings can open an avenue to find new therapeutics for burn wounds healing, anti-inflammatory and antioxidant properties.

Effects of luteolin supplementation on growth, histology, antioxidant capacity, non−specific immunity and intestinal microbiota of the red swamp crayfish (Procambarus clarkii)

To investigate the effects of luteolin (LUT) on the growth, antioxidant capacity, non−specific immunity and intestinal microbiota of the red swamp crayfish (Procambarus clarkii), 300 individual P. clarkii (4.17 ± 0.50g average body weight and 50.19 ± 1.40mm average body length) were randomly divided into five groups and fed with LUT doses of 0mg/kg (CG), 50mg/kg (LG50), 100mg/kg (LG100), 300mg/kg (LG300), and 500mg/kg (LG500) for 8 weeks, respectively. Firstly, the results demonstrated that survival rates were significantly higher in all LUT added groups compared to the CG group, with no significant differences in hepatosomatic index, feed coefficient and muscle components. Secondly, dietary supplementation with LUT improves the growth performance of P. clarkii. Thirdly, histological observations displayed that the hepatopancreas and intestine tissues were not damaged in the LUT−supplemented group, and the length of intestinal folds was significantly increased in the LG100 and LG300 groups. Fourthly, the LG50, LG100, and LG300 groups produced significantly increased hepatopancreatic antioxidant enzyme activities and hemolymph immune indicators compared with the CG group. In all experimental groups, malondialdehyde contents were significantly reduced, as were aspartate aminotransferase and alanine aminotransferase activities. Adding LUT to the diet specifically improved the structure and optimized the function of the gut microbiota. Compared to the CG group, the LG group (100mg/kg) displayed a significant increase in the index of Chao1 and Observed species, an increase in the relative abundance of Proteobacteria and Actinobacteria, a decrease in the ratio of Bacteroidetes to Firmicutes, and an increase in the abundance of Thermomonas, Turicibacter, Bdellovibrio and Enterococcus. PICRUSt analysis demonstrated a significant improvement in the KEGG pathway associated with isoflavone biosynthesis in the LG group. In conclusion, the present study confirms that dietary supplementation with 50−300mg/kg LUT favors growth, non−specific immunity and antioxidant function in P. clarkii, with optimal additions ranging from 102−165mg/kg. Furthermore, it optimized gut microbiota function, suggesting that LUT can be used as a potential feed additive for P. clarkii. These findings provide new insights for healthy P. clarkii culture.

Expression analysis of nuclear factor kappa B (NF-κB) in oral squamous cell carcinoma

IntroductionOral squamous cell carcinoma (OSCC) is the most common cancer in the head and neck region. More than 45 % of the cancer cases in India are diagnosed as squamous cell carcinoma. Although much progress has been made in controlling OSCC in recent years, the management or treatment option for OSCC remains challenging. Interestingly, nuclear factor kappa B (NF-κB) is a protein complex that controls transcription of DNA, cytokine production, and cell survival. Thus, this study aims to evaluate the levels of NF-κB in OSCC patients and their adjacent normal tissue. Materials and methodA total of 20 OSCC and adjacent normal tissues were collected from the biopsy samples of Department of Oral and Maxillofacial Surgery, Saveetha Dental College, and Hospitals. The tissues were processed for Hematoxylin and Eosin staining and NF-κB gene expression studies. The data were shown as mean ± standard deviation and p<0.05* was statistically significant. Results and conclusionOur histological observation findings showed that there was epithelial cell injury in the OSCC samples, which contributed to the creation of more keratin pearls. Additionally, our findings showed that as compared to normal tissue, OSCC samples had considerably greater NF-kB expression levels. As a result, we conclude that NF-κB was found to be an important oncogene in the tumor growth and metastasis of various cancers, including OSCC. Thus, inhibition of NF-κB can be postulated to be a potential treatment against OSCC.

Developmental characteristics of aggregated lymphoid nodules area in the abomasum of fetal Bactrian camels (Camelus bactrianus)

BackgroundBactrian camel is one of the important economic animals in northwest China. They live in arid desert, and their gestation period is about 13 months, which is longer than other ruminants (such as cattle and sheep). The harsh living conditions have made its unique histological characteristics a research focus. Aggregated lymphoid nodules area (ALNA) in the abomasum of Bactrian camels, as one of the most important sites for the induction of the immune response, provide a comprehensive and effective protective role for the organism, and their lack of information will affect the feeding management, reproduction and epidemic prevention of Bactrian camels. In this study, the histological characteristics of the fetal ALNA in the abomasum of Bactrian camels at different developmental gestation have been described by using light microscopy and histology .ResultsThe ALNA in the abomasum of the Chinese Alashan Bactrian camel is a special immune structure that was first discovered and reported by Wen-hui Wang. To further establish the developmental characteristics of this special structure in the embryonic stage, the abomasum ALNA of 8 fetuses of Alashan Bactrian camels with different gestational ages (5~13 months) were observed and studied by anatomy and histology. The results showed that the aggregation of reticular epithelial cells (RECs) surrounded by a very small number of lymphoid cells was detected for the first time in the abomasum of fetal camel at 5 months gestation, which was presumed to be primitive ALNA. At 7 months gestation, the reticular mucosal folds region (RMFR) appeared, but the longitudinal mucosal folds region (LMFR) was not significant, and histological observations showed that there were diffusely distributed lymphocytes around the RECs. At 10months gestation, RMFR and LMFR were clearly visible, lymphoid follicles appeared in histological observation, lymphocytes proliferated vigorously. By 13 months, the volume of lymphoid follicles increased, forming the subepithelial dome (SED), and there was a primitive interfollicular area between the lymphoid follicles, which contained high endothelial vein (HEV), but no germinal center (GC) was found. In summary, ALNA of Bactrian camels is not fully mature before birth.ConclusionsGenerally, the small intestine PPs of ruminants (such as cattle and sheep) is already mature before birth, while the ALNA in the abomasum of Bactrian camels is not yet mature in the fetal period. During the development of ALNA in Bactrian camel, the development of lymphoid follicles extends from submucosa to Lamina propria. Interestingly, the deformation of FAE changes with age from simple columnar epithelium at the beginning of pregnancy to Simple cuboidal epithelium, which is opposite to the FAE deformation characteristics of PPs in the small intestine of fetal cattle and sheep. These results are the basis of further research on the specificity of ALNA in the abomasum of Bactrian camels.

Ecotoxicological evaluation of chitosan biopolymer films particles in adult zebrafish (Danio rerio): A comparative study with polystyrene microplastics

To mitigate the environmental impact of microplastics (MPs), the scientific community has innovated sustainable and biodegradable polymers as viable alternatives to traditional plastics. Chitosan, the deacetylated form of chitin, stands as one of the most thoroughly investigated biopolymers and has garnered significant interest due to its versatile applications in both medical and cosmetic fields. Nevertheless, there is still a knowledge gap regarding the impact that chitosan biopolymer films (CBPF) may generate in aquatic organisms. In light of the foregoing, this study aimed to assess and compare the potential effects of CBPF on the gastrointestinal tract, gills, brain, and liver of Danio rerio against those induced by MPs. The findings revealed that both CBPF and MPs induced changes in the levels of oxidative stress biomarkers across all organs. However, it is essential to note that our star plots illustrate a tendency for CBPF to activate antioxidant enzymes and for MPs to produce oxidative damage. Regarding gene expression, our findings indicate that MPs led to an up-regulation in the expression of genes associated with apoptotic response (p53, casp3, cas9, bax, and bcl2) in all fish organs. Meanwhile, CBPF produced the same effect in genes related to antioxidant response (nrf1 and nrf2). Overall, our histological observations substantiated these effects, revealing the presence of plastic particles and tissue alterations in the gills and gastrointestinal tract of fish subjected to MPs. From these results, it can be concluded that CBPF does not represent a risk to fish after long exposure.

Bulked Segregant RNA-Seq Reveals Different Gene Expression Patterns and Mutant Genes Associated with the Zigzag Pattern of Tea Plants (Camellia sinensis).

The unique zigzag-patterned tea plant is a rare germplasm resource. However, the molecular mechanism behind the formation of zigzag stems remains unclear. To address this, a BC1 genetic population of tea plants with zigzag stems was studied using histological observation and bulked segregant RNA-seq. The analysis revealed 1494 differentially expressed genes (DEGs) between the upright and zigzag stem groups. These DEGs may regulate the transduction and biosynthesis of plant hormones, and the effects on the phenylpropane biosynthesis pathways may cause the accumulation of lignin. Tissue sections further supported this finding, showing differences in cell wall thickness between upright and curved stems, potentially due to lignin accumulation. Additionally, 262 single-nucleotide polymorphisms (SNPs) across 38 genes were identified as key SNPs, and 5 genes related to zigzag stems were identified through homologous gene function annotation. Mutations in these genes may impact auxin distribution and content, resulting in the asymmetric development of vascular bundles in curved stems. In summary, we identified the key genes associated with the tortuous phenotype by using BSR-seq on a BC1 population to minimize genetic background noise.

MiR-202-3p Targets Calm1 and Suppresses Inflammation in a Mouse Model of Acute Respiratory Distress Syndrome.

Acute respiratory distress syndrome (ARDS) is regarded as a type of respiratory failure. Emerging evidence has demonstrated the significant roles of microRNAs in various disorders. Nevertheless, the role of miR-202-3p in ARDS is unclear. Forty male C57BL/6 mice treated with phosphate buffer saline/lipopolysaccharide (PBS/LPS) and administrated with NC/miR-202-3p agomir were divided into four groups. A reverse transcription-quantitative polymerase chain reaction was used to evaluate the level of miR-202-3p, its target genes, and proinflammatory factors. Hematoxylin‑eosin was utilized for histological observation of the lung tissues. The Wet/Dry ratio, myeloperoxidase activity, and total protein concentration in bronchoalveolar lavage fluid were assessed to determine pulmonary edema. Western blotting was used for quantifying protein levels of proinflammatory factors, nuclear factor kappa B (NF-κB), and NLR family pyrin domain containing 3 (NLRP3) signaling-associated proteins. Calmodulin 1 (Calm1) protein expression in murine lung tissues was evaluated by immunohistochemistry. The binding relation between miR-202-3p and Calm1 was assessed by luciferase reporter assay. The results showed that miR-202-3p was lowly expressed in the lung tissues of ARDS mice. Overexpressed miR-202-3p relieved LPS-induced edema, reduced proinflammatory factors, and inactivated NF-κB/NLRP3 signaling in murine lung tissues. Calm1 was targeted by miR-202-3p and displayed a high level of LPS-induced ARDS. In conclusion, miR-202-3p targets Calm1 and suppresses inflammation in LPS-induced ARDS, thereby inhibiting the pathogenesis of ARDS in a mouse model.

Gene expression profiles of Japanese precious coral Corallium japonicum during gametogenesis.

Corallium japonicum, a prized resource in Japan, plays a vital role in traditional arts and fishing industries. Because of diminished stock due to overexploitation, ongoing efforts are focused on restoration through transplantation. This study aimed to enhance our understanding of the reproductive biology of these valuable corals and find more efficient methods for sex determination, which may significantly contribute to conservation initiatives. We used 12 three-month aquarium reared C. japonicum colony fragments, conducted histological analysis for maturity and sex verification, and performed transcriptome analysis via de novo assembly and mapping using the C. rubrum transcriptome to explore gene expression differences between female and male C.japonicum. Our histological observations enabled sex identification in 33% of incompletely mature samples. However, the sex of the remaining 67% of samples, classified as immature, could not be identified. RNA-seq yielded approximately 21-31 million short reads from 12 samples. De novo assembly yielded 404,439 highly expressed transcripts. Among them, 855 showed significant differential expression, with 786 differentially expressed transcripts between females and males. Heatmap analysis highlighted 283 female-specific and 525 male-specific upregulated transcripts. Transcriptome assembly mapped to C. rubrum yielded 28,092 contigs, leading to the identification of 190 highly differentially expressed genes, with 113 upregulated exclusively in females and 70 upregulated exclusively in males. Blastp analysis provided putative protein annotations for 83 female and 72 male transcripts. Annotation analysis revealed that female biological processes were related to oocyte proliferation and reproduction, whereas those in males were associated with cell adhesion. Transcriptome analysis revealed sex-specific gene upregulation in incompletely mature C. japonicum and shared transcripts with C. rubrum, providing insight into its gene expression patterns. This study highlights the importance of using both de novo and reference-based assembly methods. Functional enrichment analysis showed that females exhibited enrichment in cell proliferation and reproduction pathways, while males exhibited enrichment in cell adhesion pathways. To the best of our knowledge, this is the first report on the gene expressions of each sex during the spawning season. Our findings offer valuable insights into the physiological ecology of incompletely mature red Japanese precious corals and suggest a method for identifying sex using various genes expressed in female and male individuals. In the future, techniques such as transplantation, artificial fertilization, and larval rearing may involve sex determination methods based on differences in gene expression to help conserve precious coral resources and ecosystems.

Different Responses of Histology, Antioxidant, and Inflammation in Gill and Kidney of Yellow Catfish Pelteobagrus fulvidraco under Three Dietary Fat Levels

This experiment investigated the influences of different dietary fat levels on histology, oxidative status, and immune response in gill and kidney of yellow catfish (Pelteobagrus fulvidraco). Three diets with different fat levels of 63.1 g/kg (low-fat, LF), 93.3 g/kg (medium-fat, MF), and 153.2 g/kg (high-fat, HF) were prepared to feed yellow catfish. The experiment continued for 56 days, and at the end of the experiment, gill and kidney tissues were sampled. As a result, both gill and kidney showed different degrees of tissue damage in HF group in terms of histology observation. HF increased the malondialdehyde content in gill but showed no effect on kidney, indicating that gill is more susceptible to injury than kidney under high-energy intake conditions. Additionally, HF diet significantly increased the activities of total-superoxide dismutase and catalase to eliminate excess peroxides both in gill and kidney. Moreover, HF diet significantly upregulated the expressions of pro-inflammatory cytokines (il6 and tnfα) and downregulated the expression of anti-inflammatory cytokines (il10), indicating that HF-diet-induced inflammatory response both in gill and kidney. These findings reveal the potential regulatory approach for fish gill and kidney health by dietary fat level, which will help to understand the adverse impacts of dietary lipid imbalance on the health of fish.

Thymic Mimetic Cells: Ontogeny as Immunology.

Medullary thymic epithelial cells (mTECs) generate immunological self-tolerance by ectopically expressing peripheral-tissue antigens (PTAs) within the thymus to preview the peripheral self to maturing Tcells. Recent work, drawing inspiration from old histological observations, has shown that subtypes of mTECs, collectively termed mimetic cells, co-opt developmental programs from throughout the organism to express biologically coherent groups of PTAs. Here, we review key aspects of mimetic cells, especially as they relate to the larger contexts of molecular, cellular, developmental, and evolutionary biology. We highlight lineage-defining transcription factors as key regulators of mimetic cells and speculate as to what other factors, including Aire and the chromatin potential of mTECs, permit mimetic cell differentiation and function. Last, we consider what mimetic cells can teach us about not only the thymus but also other tissues.

Annual and lunar-cycle-related breeding rhythmicity in the acorn barnacle Tetraclita kuroshioensis Chan, Tsang &amp; Chu, 2007 (Cirripedia: Balanomorpha: Tetraclitidae) in Okinawa, Japan

Abstract We investigated the reproduction of the barnacle Tetraclita kuroshioensisChan, Tsang &amp; Chu, 2007 (Cirripedia: Thoracica: Tetraclitidae) at different heights in the intertidal zone in Okinawa, Japan with a focus on gonadal development, the brooding of eggs, and unhatched larvae, as well as the role of environmental factors in breeding rhythmicity. Histological observations collected around each full moon from September 2019 to October 2020 showed various stages of oocyte development in September and October 2019 and from March to September 2020, whereas mature testes were observed from April to October 2020. The breeding season of this species thus lasts for at least seven months under long-day conditions at a water temperature above 21 °C. Barnacles collected from three tidal levels on multiple occasions during April, August, and October 2020 displayed little vertical variation in their stage of ovarian development, but there was a noticeable difference in the percentage of brooding individuals in August and October, as well as an increased prevalence of brooding during the new-moon phase at the upper and middle tidal heights. Our findings indicate that larval release by T. kuroshioensis undergoes seasonal variation, apparently influenced by the cycles of lunar (major factor) and tide (minor factor) in their habitat.

Morphological characteristics of sex dimorphism in Gobiopterus sp. (Gobiiformes: Oxudercidae) from Ranu Grati Lake, Pasuruan District, East Java, Indonesia

Abstract. Anitasari S, Arfiati D, Susilo, Marhendra APW. 2024. Morphological characteristics of sex dimorphism in Gobiopterus sp. (Gobiiformes: Oxudercidae) from Ranu Grati Lake, Pasuruan District, East Java, Indonesia. Biodiversitas 25: 1214-1222. Ranu Grati is a volcanic lake located in Pasuruan, East Java, Indonesia which has high ecological value as a habitat for a unique endemic fish found only in this lake, namely the Lempuk fish (Gobiopterus sp.). The aim of this research is to determine the morphological characters based on sexual dimorphism of Lempuk fish. in Ranu Grati Lake. The result showed the body color of male and female Lempuk fish is transparent. Lempuk fish do not have sexual dichromatism or their sex cannot be distinguished based on their body color. Lempuk fish have vomer teeth and canine teeth. Types of male and female ctenoid scales have a comb-like shape. The position of the mouth of the Lempuk Fish is located near the top or head of the fish but not precisely at the tip. There are variations in the superior subterminal mouth type between male and female fish, especially in terms of anatomical details and mouth structure. Based on meristic observations of the Lempuk fish, it can be explained that this fish has a fusiform body shape, this shows that gobies generally have a small fusiform body and also has pectoral fins, pelvic fins, dorsal fins, tail fins and anal fins. In this study, we found from histological observations of the Lempuk fish, mature oocytes and spermatids were found in one histological tissue sample at male fish with gonad maturity level II. This change from female to male occurs at the size of the fish with fish size of 21.21-30.58 ± 2.14mm. It indicates that is Lempuk fish is a species hermaphrodite protogyn.

The calcaneofibular ligament groove at the inferior fibula, an ultrasonographic anatomical landmark.

Calcaneofibular ligament (CFL) injuries are harder to diagnose than anterior talofibular ligament (ATFL) ones. This study aimed to clarify the fibular attachment of the CFL and verify the bony landmark for evaluating the CFL on ultrasonography. Fifty-nine ankles were used in this anatomical study. To confirm the control function of the CFL, we performed passive movement manually using cadaveric ankles and observed the ankle positions where the CFLs were tense. Histological observation of CFL attachment of the fibula was performed using Masson's trichrome stain. The ATFL and CFL were removed, and the bone morphology of the CFL attachment and inferior fibular end was imaged using a stereomicroscope and a 3D scanner. Using ultrasonography, we evaluated the bone morphology of the fibular attachment of the CFL in short-axis images of 27 healthy adult ankles. The CFL was tensed according to ankle motions: supination, maximum dorsi flexion, maximum plantar flexion, and mild plantar flexion-external rotation. Below the CFL attachment of the fibula was a slight groove between the inferior tip and the obscure tubercle of the fibula. This groove was observed in 81.5% of cases using short-axis ultrasonography. The CFL was tensed in various ankle positions to control the movements of the talocrural and subtalar joints. There was a slight groove at the inferior end of the fibula where the CFL coursed downward. We called it the CFL groove and proposed that it could serve as a landmark for the short-axis image of ultrasonography.

A novel magnetic compression technique for establishment of a vesicovaginal fistula model in Beagle dogs

Vesicovaginal fistula lacks a standard, established animal model, making surgical innovations for this condition challenging. Herein, we aimed to non-surgically establish vesicovaginal fistula using the magnetic compression technique, and the feasibility of this method was explored using eight female Beagle dogs as model animals. In these dogs, cylindrical daughter and parent magnets were implanted into the bladder and vagina, respectively, after anesthesia, and the positions of these magnets were adjusted under X-ray supervision to make them attract each other, thus forming the structure of daughter magnet-bladder wall-vaginal wall-parent magnet. Operation time and collateral damage were recorded. The experimental animals were euthanized 2 weeks postoperatively, and the vesicovaginal fistula gross specimens were obtained. The size of the fistula was measured. Vesicovaginal fistula was observed by naked eye and under a light microscope. Magnet placement was successful in all dogs, and remained in the established position for the reminder of the experiment. The average operation time was 14.38 min ± 1.66 min (range, 12–17 min). The dogs were generally in good condition postoperatively and were voiding normally, with no complications like bleeding and urine retention. The magnets were removed from the vagina after euthanasia. The vesicovaginal fistula was successfully established according to gross observation, and the fistula diameters were 4.50–6.24 mm. Histological observation revealed that the bladder mucosa and vaginal mucosa were in close contact on the internal surface of the fistula. Taken together, magnetic compression technique is a simple and feasible method to establish an animal model of vesicovaginal fistula using Beagle dogs. This model can help clinicians study new surgical techniques and practice innovative approaches for treating vesicovaginal fistula.

Pioneering a chick embryo model to explore the intrauterine etiology of developmental dysplasia of the hip in oligohydramnios conditions

ObjectiveTo explore the impact of oligohydramnios on fetal movement and hip development, given its association with Developmental Dysplasia of the Hip (DDH) but unclear mechanisms. MethodsChick embryos were divided into four groups based on the severity of oligohydramnios induced by amniotic fluid aspiration (control, 0.2 mL, 0.4 mL, 0.6 mL). Fetal movement was assessed by detection of movement and quantification of residual amniotic fluid volume. Hip joint development was assessed by gross anatomic analysis, micro-computed tomography (micro-CT) for cartilage assessment, and histologic observation at multiple time points. In addition, a subset of embryos from the 0.4 mL aspirated group underwent saline reinfusion and subsequent evaluation. ResultsIncreasing volumes of aspirated amniotic fluid resulted in worsening of fetal movement restrictions (e.g., 0.4mL aspirated and control group at E10: frequency difference -7.765 [95% CI: -9.125, -6.404]; amplitude difference -0.343 [95% CI: -0.588, -0.097]). The 0.4mL aspirated group had significantly smaller hip measurements compared to controls, with reduced acetabular length (-0.418 [95% CI: -0.575, -0.261]) and width (-0.304 [95% CI: -0.491, -0.117]) at day E14.5. Histological analysis revealed a smaller femoral head (1.084±0.264 cm) and shallower acetabulum (0.380±0.106 cm) in the 0.4mL group. Micro-CT showed cartilage matrix degeneration (13.6% [95% CI: 0.6%, 26.7%], P=0.043 on E14.5). Saline reinfusion resulted in significant improvements in the femoral head to greater trochanter (0.578 [95% CI: 0.323, 0.833], P=0.001). ConclusionsOligohydramnios can cause DDH by restricting fetal movement and disrupting hip morphogenesis in a time-dependent manner. Timely reversal of oligohydramnios during the fetal period may prevent DDH.