Intensity Of Expression Research Articles

De Novo Synthesis of Reticuline and Taxifolin Using Re-engineered Homologous Recombination in Yarrowia lipolytica.

Yarrowia lipolytica has been widely engineered as a eukaryotic cell factory to produce various important compounds. However, the difficulty of gene editing and the lack of efficient neutral sites make rewiring of Y. lipolytica metabolism challenging. Herein, a Cas9 system was established to redesign the Y. lipolytica homologous recombination system, which caused a more than 56-fold increase in the HR efficiency. The fusion expression of the hBrex27 sequence in the C-terminus of Cas9 recruited more Rad51 protein, and the engineered Cas9 decreased NHEJ, achieving 85% single-gene positive efficiency and 25% multigene editing efficiency. With this system, neutral sites on different chromosomes were characterized, and a deep learning model was developed for gRNA activity prediction, thus providing the corresponding integration efficiency and expression intensity. Subsequently, the tool and platform strains were validated by applying them for the de novo synthesis of (S)-reticuline and (2S)-taxifolin. The developed platform strains and tools helped transform Y. lipolytica into an easy-to-operate model cell factory, similar to Saccharomyces cerevisiae.

Individual Genomic Loci, Transcript Level and Serum Profile of Immune, Antioxidant and Hormonal Markers Associated with Sheep Arthritis

Arthritis is a leading cause of economic loss in livestock farming including sheep. This study examined the changes in gene expression, antioxidants, pro-inflammatory cytokines, acute-phase proteins (APPs), hormonal assays and iron profiles linked to sheep arthritis, as well as the diagnostic utility of these markers. Blood samples were obtained from 30 apparently healthy rams and 30 rams with arthritis for gene expression and biochemical analyses. Gene expression intensities were much higher in the arthritis-affected rams than in the healthy ones for the genes IL-1α, IL-1β, IL-6, IL-10, TNFα, NCF4, NFKB, TMED, FCAMR, iNOS and COX18. The SOD3, CAT, GPX and ATOX1 genes were expressed at substantially lower levels in arthritis-affected rams. Disparities in the nucleotide sequence variants for the amplified DNA bases linked to arthritis for the studied genes were found in the PCR-DNA sequence verdicts of the affected and healthy rams. Immunological, acute-phase protein (APP), antioxidant, hormonal and iron profiles were estimated in both groups and statistically analyzed. The arthritic group in relation to the healthy one showed a significant (p < 0.05) increase in pro-inflammatory cytokines, APPs, free radicals, immunoglobulins, cortisol, GH, TSH, ferritin, TIBC and UIBC and a significant (p ˂ 0.05) decrease in anti-inflammatory cytokines, antioxidants, complements, insulin, T3, T4, SI, and Tf and Tf sat.% serum levels. The estimated pro-inflammatory cytokines and APPs achieved high values of sensitivity and specificity, positive predictive values (PPVs), negative predictive values (NPVs), a high accuracy rate and a moderate likelihood ratio (LR). The study concluded that ovine arthritis stimulates innate and humeral immunity, resulting in prominent alterations in gene expression, pro-inflammatory cytokines, APP assays and antioxidant profiles, which could be valuable indicators of sheep arthritis.

Exploring Nectin-4 expression in hepatocellular carcinoma.

632 Background: Nectin cell adhesion protein 4 (Nectin-4) is overexpressed in various malignancies, contributing to cancer progression and poor prognosis. We aimed to assess Nectin-4 expression in hepatocellular carcinoma (HCC). Methods: Patients who underwent primary liver tumor resection for HCC upon initial diagnosis at National Taiwan University Hospital in 2019 were included. Exclusion criteria consisted of prior systemic or loco-regional therapy, or death within three months post-surgery. Immunohistochemical (IHC) staining for Nectin-4 (EPR15613-68, Abcam) was conducted and independently reviewed by two pathologists. Nectin-4 expression patterns (membranous, cytoplasmic, and nuclear) and intensity were scored on a 0–3 scale, with normal eccrine glands (2+) serving as a positive control. Nectin-4 expression was further assessed in primary and paired progressive or metastatic tissue samples from patients with available paired samples. Results: Of the 193 patients, 175 had specimens available for IHC analysis. Nectin-4 expression was detected in 22.9% of these patients, with intensity scores of 0 in 77.1%, 1 in 22.3%, and 2 in 0.6%. The majority of staining pattern was cytoplasmic (94.3%), with membranous staining observed in 5.1% of cases. Nectin-4 expression was significantly associated with higher pathological grades ( p = 0.005) but showed no correlation with recurrence-free survival ( p = 0.771) or overall survival ( p = 0.396) after surgery. Importantly, Nectin-4 expression intensity significantly increased in paired progressive or metastatic samples compared to initial HCC specimens ( p = 0.046; n = 27). Conclusions: Nectin-4 expression is associated with higher pathological grades but not related to clinical outcomes in HCC. Increased Nectin-4 expression in progressive or metastatic disease highlights its potential as a therapeutic target.

A practical distribution pattern of α-SMA-positive carcinoma associated fibroblasts indicates poor prognosis of patients with pancreatic ductal adenocarcinoma.

Purpose The present study aimed to clarify the distribution pattern of carcinoma associated fibroblasts (CAFs) across pancreatic ductal adenocarcinoma (PDAC) and its prognostic prediction value. Methods Data of two cohorts were retrospectively collected from consecutive patients who underwent primary pancreatic resection from January 2015 to December 2017. We used tumor specimens to screen out the most suitable markers for the spatial distribution analysis for CAFs subpopulations. We utilized a tissue microarray to assess the spatial intensity of α-SMA expression within the tumor microenvironment. Specifically, we classified CAFs into two types based on their α-SMA spatial expression. Type II CAFs were designated as those located in the juxtatumoural stroma with α-SMA expression that was moderate or higher, and those in the peripheral stroma with α-SMA expression that was less than moderate. All other cases, where the α-SMA expression did not meet these criteria, were categorized as Type I CAFs. Multivariable Cox proportional hazards regression was used to assess risk factors associated with patient outcomes. RNA sequencing data were obtained from bulk tumor samples and isolated CAFs from patients to reveal the distinct pattern and elucidated their fundamental characteristics. Results The α-SMA spatial intensity was the most suitable variable for representative of CAFs spatial characteristics. Patients with Type Ⅰ CAFs were more likely to be allocated into N1 or N2 of the N stage and Ⅱ and Ⅲ of the TNM stage. The spatial distribution pattern of CAFs (Type Ⅰ v.s. Type Ⅱ: HR, 1.568; 95 % CI, 1.053-2.334; P = 0.027) was an independent prognostic factor in the discovery cohort, so as in the validation (Type Ⅰ vs. Type Ⅱ: HR, 2.197; 95 % CI, 1.410-3.422; P = 0.001). RNA sequencing analysis revealed that the differentially expressed genes (DEGs) in Type I CAFs are closely associated with those in corresponding tumor tissues, highlighting the enhanced biological significance of immune-related and oncogenic invasive pathways. Conclusions Our findings that two types of α-SMA-positive CAFs with different spatial patterns present heterogeneously across tissues of PDACs and correlated with patients' outcomes. The spatial location of CAFs may facilitate patients' selection in precision medicine of PDACs.

Effect of electroacupuncture on vascular remodeling in rats with cerebral ischemia by regulating irisin based on VEGF/Akt/eNOS signaling pathway.

This study aimed to explore the cumulative effects and expression patterns of electroacupuncture (EA) on irisin secretion, observe the effects of EA on the recovery of neurobehavioral function and vascular remodeling after cerebral ischemia, and elucidate the mechanism by which EA promotes vascular remodeling by regulating irisin expression. A rat model of left middle cerebral artery occlusion (MCAO) was prepared, and EA was performed. Tissue distribution and expression of irisin were determined by immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and Western blotting. Type III fibronectin domain protein 5-silenced adeno-associated virus (rAAV-shFNDC5) was injected into the lateral ventricle as a control. Neurobehavioral function was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining and behavioral experiments, while vascular remodeling was evaluated using laser speckle blood flow imaging, and the expressions of irisin and vascular remodeling-related factors were measured by ELISA and Western blotting. The number of FNDC5-positive neurons, fluorescence intensity, and irisin expression reached their maximum increase after 7 days of EA treatment. In addition, the EA group exhibited a significant reduction in cerebral infarct volume and impairment of neurobehavioral function, an increase in cerebral blood flow and microvascular diameter on the ischemic side, and significantly higher expression levels of FNDC5, brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), protein kinase B (Akt), and endothelial nitric oxide synthase (eNOS). However, rAAV-shFNDC5 significantly weakened the therapeutic effects of EA. EA upregulated irisin expression, reaching a peak after 7 days of EA and then stabilizing. EA facilitated vascular remodeling after cerebral ischemia, and this might be associated with the activation of the irisin-mediated VEGF/Akt/eNOS signaling pathway.

Nonverbal expressions of shame predict suicidal ideation among rurally-situated, but not urban situated, lesbian, gay, bisexual, transgender, and queer (LGBTQ) adults

In the United States (US), lesbian, gay, bisexual, transgender, and queer (LGBTQ) people experience disproportionate rates of suicidality associated with minority stress. This study aimed to investigate whether nonverbal expressions of experienced stigma (i.e., shame) predicted suicidal ideation among LGBTQ individuals with a focus on location-based disparities (comparing those living in a more rural setting to those living in a more urban setting). More specifically, we examined whether nonverbal expressions of shame predicted suicidal ideation three months later and whether this relationship was moderated by region. LGBTQ individuals (N = 133) from one rurally-situated and one urban location were videorecorded while talking about a time they felt bad about their LGBTQ identity in an observational, prospective (two-time point) design. Recordings were coded for the intensity of nonverbal expressions of shame (shoulders slumped, chest narrowed). Participants also completed several self-report measures including state shame and suicidal ideation at both the time of the recording and three months later. Moderation analyses revealed that for LGBTQ adults living in more rural settings, nonverbal shame, predicted increased suicidal ideation three months later (Bstd = 0.64, p = .005), and this was not the case for those living in more urban settings (Bstd = -0.08, p = 0.70). Self-reported shame did not predict suicidal ideation for LGBTQ adults from more rural or urban locations. These findings highlight the importance of recognizing nonverbal cues in context (i.e., in locations with more structural stigma) when assessing mental health risks and when shaping interventions for LGBTQ populations.

SOX-9 as a prognostic marker in gastric adenocarcinoma.

SRY-box transcription factor 9 (SOX9) has been reported to be overexpressed in a wide variety of gastrointestinal malignancies. While its role has been studied in gastric cancer (GC), the results remain conflicting. This study aimed to evaluate the relationship between SOX9 immunohistochemistry results and the pathological and clinical characteristics of gastric adenocarcinoma, assessing its potential as a prognostic marker. Gastric tissue samples from 150 patients with gastric cancer were included in the study. Tissue sections were stained using an anti-SOX9 antibody, and relevant data were retrospectively collected from digital records. Immunostaining results were scored based on the proportion and intensity of stained nuclei throughout the tumor. A final immunostaining score was calculated by multiplying the SOX9 intensity score by the proportion score. Strong SOX9 nuclear staining was observed in 68 patients (45.3%), while moderate staining was seen in 60 patients (40%). SOX9 nuclear staining was absent in three patients (2%). A final SOX9 immunostaining score of ≥10, classified as high expression, was identified in 60 patients (40%). Patients with higher SOX9 expression or strong intensity scores exhibited significantly larger tumor sizes, higher rates of perineural and vascular invasion, more advanced T or lymph node staging, and greater likelihoods of lymphatic or distant metastases compared to those with lower SOX9 expression or intensity scores (all P < 0.05). These findings suggest that SOX9 staining intensity and expression are associated with increased tumor malignancy and disease progression. Therefore, SOX9 may serve as a prognostic pathological indicator in GC patients.

P-37 MEDIASTINAL PARAGANGLIOMA AS A CAUSE OF HYPERTENSION IN A YOUNG PATIENT:A CASE REPORT

Abstract Introduction Paragangliomas, tumors that arise from extraadrenal chromaffin cells, are rarely found in the mediastinum. It is important to diagnose and remove paragangliomas because of its critical symptoms such as hypertension and can be curable with resection. Clinical Case A 18-year-old man, has no known disease, presented with episodes of palpitations, sweating associated with hypertension.The patient’s blood pressure measured at office was 180/110 mmHg. Electrocardiographic monitoring showed only nonspesific ST-T waves changes. Other secondary causes of hypertension were excluded. Suspecting of pheochromocytoma, 24-hour urinary metanephrine and normetanephrine levels were examined, the normetanephrine level was found to be elevated (1367,78 mcg/24 hour; normal range 88-444), whereas metanephrine level was normal (102,16 mcg/24 hour; normal range 52-341). Plasma metanephrine level was normal (58 ng/L; normal range 0-90).However plasma normetanephrine level was elevated (163,1 ng/L; normal range 0-90).The ratio of serum aldosterone level to the plasma renin activity was normal. 1 mg dexamethasone suppression test was normal. There was no adrenal mass in the abdominal MRI, but a mass measuring 4,3x 3,2 cm was seen in the right paravertebral area of the thorax in the area included in the cross-sections. In Ga-68 Dota-tate positron emission tomography (PET),a mass lesion of 43x30 mm in size, showing intense somatostatin receptor expression (SUV-max: 31.9) was detected in the right paravertebral area at the level of the 8th thoracic vertebra, which was evaluated as paraganglioma. For more than 1 month, the patient had been prepared for the surgery administering combination doxazosine and propronol after the treatment with only doxazosin. The patient was consulted to the thoracic surgery department. Phentolamine was provided before surgery. The mass was resected via thoracotomy. In histological evaluation, widespread zelballen and pseudorosette structures were observed. There was no comedonecrosis, and there was no vascular and capsular invasion. Ki 67 proliferation index was 4-5 %. On immunohistochemical staining, the mass tested positive for synaptophysin and chromagranin and negative for cytokeratins. One month after surgery normetanephrine (147,85 mcg/24h; normal range: 88-444) and metanephrine (102,16 mcg/24 h; normal range 52-341) levels were normal. The patients blood pressure was stabilized at 120/70 mmHg and heart rate was 80 bpm. In the genetic analysis of the patient, the NDUFA6 gene was detected and mitechondrial complex 1 deficiency was associated with nuclear type 33 disease and the patient was accepted as a carrier. Screening with 24 hour urine metanephrine and normetanephrine was recommended to patient’s family members. Conclusion Since, there are no definite microscopic criteria for the distinction between benign and malignant tumors, radical excision is necessary. The symptoms are curable with resection of tumor, long term follow up for recurrence is important.Figure 1:Gallium-68 Dota-Tate PETShowing the mass(SUV-max:31.99)

Expression of TNF-α, VEGF-A and Microvessel Density in Cerebral Alveolar Echinococcosis and Their Correlation with Perilesional Brain Edema.

Alveolar echinococcosis (AE) is an infrequent zoonosis caused by Echinococcus multilocularis with a high degree of disability and mortality. Metastatic cerebral alveolar echinococcosis (CAE) is very rare and the lesions could lead to severe perilesional brain edema (PLBE) and subsequent uncontrollable intracranial hypertension. In this study, we sought to determine the expression of edema-associated factors in CAE lesions and their associations with PLBE. We retrospectively evaluated the clinical data of 18 CAE patients who received craniotomy. Severity of PLBE was described by edema index (EI). Archived specimens were processed for immunohistochemistry to detect tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor A (VEGF-A) and microvessel density (MVD) in CAE lesions. Expression intensity of CAE lesions was quantified by integral optical density (IOD) or count and was compared to the control group. The results showed TNF-α and VEGF-A were significantly expressed in CAE lesions (p < 0.001), their levels were positively correlated with PLBE (TNF-α: r = 0.701, p = 0.001; VEGF-A: r = 0.803, p < 0.001). The MVD of CAE lesions had a similar expression with normal brain tissue, and it was positively correlated with PLBE and VEGF-A (PLBE: r = 0.849, p < 0.001; VEGF-A: r = 0.687, p = 0.002). In conclusion, we speculated the upregulation of TNF-α and VEGF-A induced the formation of PLBE. Besides, though there was no extra increase of MVD, it was still regulated by VEGF-A and provided a better anatomical basis for the formation of PLBE and further promoted it.

Role of Dynamic Facial Expressions in Judgement of Attractiveness: AI and Human Analysis

Facial attractiveness plays a crucial role in social interactions and mate selection, influencing judgments and behaviors in various contexts. While extensive research has examined structural facial features such as symmetry and averageness, the dynamic and modifiable nature of facial expressions remains underexplored. This study explores the influence of dynamic, modifiable facial expressions on perceived attractiveness through two experiments utilizing both human raters and artificial intelligence (AI) to analyze the relationship between facial expression positivity and attractiveness. In Experiment 1, we examined the effects of the experimenter’s control over the facial expressions on the facial expression and attractiveness ratings using two types of videos: one in which the stimulus models introduced themselves and another in which the subjects were controlled to be expressionless. Results revealed that the positivity and intensity of facial expressions, as assessed by facial expression analysis software, fluctuated based on the experimenter’s manipulation. Additionally, the variation in raters’ evaluations of facial expression positivity led to corresponding changes in attractiveness ratings. In Experiment 2, we investigated the relationship between facial expression ratings by humans and AI and their connection to attractiveness. The positivity of facial expressions in the video stimuli was analyzed by students and an AI-based facial expression analysis software, whereas a separate group of students rated the attractiveness of the same stimuli. Findings showed a positive correlation between the attractiveness ratings for both male and female stimuli and the positivity of facial expressions rated by both humans and AI. This indicates that attractive faces were perceived as having positive expressions and the actual positivity of the expressions themselves influenced attractiveness evaluations. These findings indicate that positivity in facial expressions plays a significant role in attractiveness perception. Future research should investigate how the relationship between attractiveness and expressions may be shaped by evolutionary pressures and social behaviors.

Immunohistochemical analysis of 147 cases of low-grade endometrial stromal sarcoma: refining the immunohistochemical profile of LG-ESS on a large, molecularly confirmed series.

Low-grade endometrial stromal sarcoma (LG-ESS) can present diagnostic challenges, due to its overlapping morphological features with other uterine mesenchymal tumors. Misdiagnosis rates remain significant, and immunohistochemical data for LG-ESS are limited to small series and inconsistent antibody panels. This study aimed to refine the IHC profile of LG-ESS by analyzing a large, molecularly confirmed series of 147 cases using a panel of 24 antibodies, including newer markers like transgelin and smoothelin. CD10 and IFITM1, key endometrial stromal markers, were expressed in 86% (92% of those extensively) and 69% (60% of those extensively) of cases, with fusion-positive tumors showing significantly higher expression. Smooth muscle markers (α-SMA, desmin, h-caldesmon, calponin, transgelin) were variably expressed, predominantly in focal or low-intensity patterns, with α-SMA reaching the highest frequency of expression (44%). However, the intensity of smooth muscle marker expression was usually very low. Smoothelin was rarely expressed. Hormone receptors were frequently positive, with PR showing a higher frequency (92% vs. 83%) and intensity than ER. Markers like S-100, HMB45, and CD117 were largely negative; all tumors were p53 wild-type, with preserved SMARCB1/SMARCA4 expression and ALK and ROS1 negativity. This work represents the largest molecularly validated IHC study on LG-ESS, providing a robust diagnostic profile for routine pathology. By addressing key diagnostic limitations and examining newer markers, our study supports a more standardized approach to diagnosing LG-ESS and underscores the value of immunohistochemical panels, particularly in fusion-negative tumors where diagnosis relies on morphological and immunohistochemical interpretation. These findings contribute critical data for improving diagnostic accuracy.

Intense FAP Expression of Ovarian Metastatic Breast Cancer Detected by [68Ga]RTX-1363 PET/CT.

PET/CT targeting fibroblast activation protein α (FAP) in cancer-associated fibroblasts shows promise in theranostics. Here, we report the case of a 31-year-old woman with hormone receptor-positive/human epidermal growth factor receptor 2-negative breast cancer who presented with rising CA15-3 for further diagnostic workup. Whereas [18F]FDG PET/CT was unremarkable, novel [68Ga]RTX-1363 PET/CT revealed intense tracer accumulation in thoracoabdominal lymph nodes and both ovaries. Follow-up imaging confirmed tumor progression, and diagnostic laparoscopy verified metastatic disease in the ovaries with high FAP expression in the tumor stroma. This case underscores the superior sensitivity of [68Ga]RTX-1363 PET/CT over [18F]FDG PET/CT, enhancing breast cancer diagnostic and therapeutic strategies.

The role of OCT4 and CD44 in lower lip carcinogenesis.

Carcinogenesis is characterized by the transformation of normal cells into malignant cells. Concerning the lower lip, exposure to UV radiation is the main etiological factor associated with the development of epidermoid carcinomas and actinic cheilitis. According to the hierarchical model theory, cancer development is driven by populations of cancer stem cells. In this context, this study aimed to compare the expression of octamer-binding transcription factor 4 (OCT4) and CD44 in 40 lower lip epidermoid carcinoma (LLEC) and 40 actinic cheilitis (AC) cases. OCT4 and CD44 expressions were assessed semi-quantitatively according to the percentage of positive epithelial cells (PP) and intensity of expression (IE), resulting in a total immunolabeling score (PIT). No statistically significant differences were detected between OCT4 and CD44 immunoexpression and clinicopathological parameters, except for lymph node metastasis, in which a decrease in CD44 expression in the core tumor of cases with lymph node metastasis was observed. Furthermore, decreased CD44 expression was observed in LLEC cases when compared to AC cases. The findings reported herein suggest a higher participation of CD44 in early carcinogenesis stages. In addition, the imbalance between OCT4 and CD44 immunoexpressions suggests the presence of different neoplastic cell subpopulations.

Evaluation of the effects of carbamazepine-loaded chitosan-coated PLGA-Zein nanoparticles on pilocarpine-induced seizure model in zebrafish larvae: Developmental toxicity and behavioral assays.

Epilepsy, the most common neurological disorder worldwide, is characterized by sudden paroxysmal brain activity, which can be generalized or focal. Extensive research has explored various treatment strategies for this condition. Our study employed a pilocarpine (PL)-induced seizure model in zebrafish (Danio rerio) embryos and larvae to assess the effects of carbamazepine (CBZ)-loaded chitosan-coated PLGA-Zein nanoparticles (NPs) over 96 hours. We evaluated the developmental toxicity (mortality, malformation, and larval hatching), behavioral changes (sensorimotor reflexes), and histopathological and immunohistochemical alterations in brain tissue, focusing on 5-hydroxytryptamine receptor 4 (5HT4), and brain and muscle ARNT-Like 1 (BMAL1) expressions. Our findings revealed high mortality and malformation rates in groups treated with pure CBZ (PL+CBZ 50 and PL+CBZ 100). These groups also exhibited delayed hatching and impaired sensorimotor reflexes. In contrast, the CBZ-NP-treated groups (PL+CBZ NP 50 and PL+CBZ NP 100) showed hatching rates comparable to the control group, with significantly lower mortality and malformation rates compared to pure CBZ-treated groups. Moreover, intense cytoplasmic expression of 5HT4 and BMAL1 was observed in neuropils of the PL+CBZ 100 group. This study highlights the potential of CBZ-loaded NPs in reducing developmental toxicity and adverse neurological effects associated with pure CBZ treatment in seizure models.

Karnozin EXTRA® causes changes in mitochondrial bioenergetics response in MCF-7 and MRC-5 cell lines.

Numerous studies reported about potential effects of L-carnosine in regulation of tumor growth and metabolism. We evaluated the effects of different concentrations of L-carnosine from Karnozin EXTRA® supplement on mitochondrial respiratory chain complexes of human embryo lung fibroblasts (MRC-5) and human breast cancer cells (MCF-7), with different energy pathways. Also, we analyzed the proliferation index and expression of various markers of oxidative stress. Treatment with Karnozin EXTRA® (concentration of L-carnosine were 2, 5 and 10mM) for 24hours gradually decreased the number of cells and changed their morphological features. In both cell lines, a dose-dependent reduction of cell viability was recorded compared to the control group. Also, experimental groups showed a concentration-dependent decrease in fluorescence intensity of SOD2 expressions in MCF-7, while in MRC-5 we noticed higher fluorescence intensity in Carnosine 2mM group. Treated cells, in both cell lines, showed different intensity of iNOS cytoplasmic immunopositivity in a concentration-dependent manner. In all experimental groups, we noticed an increased expression of marker of oxidative stress-cytochrome P450 2E1 (CYP2E1). The effects of Karnozin EXTRA® capsule on mitochondrial respiration, assessed with the Clark-type electrode, were manifested as a reduction of: basal cell respiration, maximum capacity of electron transport chain and mitochondrial ATP-linked respiration. Also, significant decrease in the activity of complex I (NADH-ubiquinone oxidoreductase), complex II (succinate dehydrogenase) and complex IV (cytochrome c oxidase) was observed in both cell lines. Bearing in mind that Karnozin EXTRA® is a potential regulator of energy metabolism of MCF-7 and MRC-5, these results provide a good basis for further preclinical and clinical research.

Dominant Expression of Chromogranin B in Pituitary Corticotrophs and Its Putative Role in Interaction With Secretogranin III.

SummaryPrevious studies have suggested that chromogranin A (CgA) is a partner molecule of secretogranin III (SgIII). In mouse pituitary corticotroph-derived AtT-20 cells, SgIII plays a role in sorting CgA/hormone aggregates into secretory granules (SGs). Although CgA expression is equivocal, CgB is clearly detectable in the rat pituitary corticotrophs. Therefore, we hypothesized that CgB shares a function with CgA in pituitary corticotrophs. In the binding assays, CgB, similar to CgA, showed binding activity to SgIII under weakly acidic conditions and in the presence of Ca2+. Considering the differences in animal species, the different abilities of antibodies, and the conditions of tissue fixation and thin sectioning in immunofluorescence histochemistry, we found that CgA was expressed in a small population (approximately 10%), and its expression intensity was weaker than that of CgB (>98%) in rodent pituitary corticotrophs. In addition, similar to CgA, CgB and SgIII were colocalized in adrenocorticotropic hormone (ACTH) granules. The labeling of CgA and CgB was not completely consistent, and CgB colocalized with SgIII in many granules. These results suggest that there are multiple sorting systems for ACTH granules in pituitary corticotrophs and that the SgIII/CgB complex behaves more dominantly than the SgIII/CgA complex, which has somewhat different properties.

Modeling and Data Analysis of Bistatic Bottom Reverberation from a Towed Horizontal Array

The spatial-temporal structures of bottom reverberation are associated with seafloor features. In a bistatic bottom reverberation experiment involving a vertical transmitting array and a towed horizontal receiving array, stable stripe structures were observed within the beam-time domain. In this study, a bistatic reverberation model based on ray theory is presented to interpret the experimental phenomena. The conventional empirical scattering function is primarily applicable to small grazing angles. Moreover, the regional segmentation method simulates reverberations across various receiving beams, ignoring scatterers in other areas. To address these issues, we substitute the empirical scattering function with a small-slope approximation (SSA) that is appropriate for full grazing angles. Furthermore, we utilize the beam pattern of arrays to incorporate the effects of each scatterer, and derive the expression for bottom reverberation intensity in both the array and beam domains. The established model demonstrates its applicability in simulating and interpreting the stripe structures of bottom reverberation, and the comparison shows that the model outputs are in agreement with the experimental results. The analysis indicates that the vertical stripes within the structures originate from eigenrays in the mirror reflection direction. Furthermore, the convex stripes are predominantly affected by the direct ray and the surface reflection ray among the scattered eigenrays, whereas the concave and elliptical stripes are primarily affected by the bottom-surface reflection ray and the surface-bottom-surface reflection ray within the scattered eigenrays.

Relative increase in walking activity in lactating dairy cows submitted to timed artificial insemination: Risk factors and subsequent fertility.

The objectives of this study were: (i) to evaluate the associations between postpartum health disorders, relative increase in walking activity (RIWA) and expression of behavioral estrus intensity captured by pedometers; (ii) to assess associations between RIWA and pregnancy in cows submitted to timed artificial insemination (TAI). Lactating dairy cows (n = 881) were enrolled in the study. Cows were submitted to a 5d TAI protocol with insertion of a progesterone device. Pregnancy status was determined by ultrasonography on D30 after AI. During the postpartum period cows were categorized as healthy (n = 468), having a reproductive disorder (n = 185), metabolic disorder (n = 118), reproductive and metabolic disorders (n = 56), or clinical mastitis (n = 54). Healthy cows tended to have (P = 0.10) a greater RIWA and a lower proportion of healthy cows did not exhibit estrus (P = 0.03) compared with cows that had a postpartum health disorder. Similarly, ovarian status at TAI protocol commencement (P = 0.01), presence of twin pregnancy (P = 0.03), and season (P < 0.0001) were all associated with RIWA. Cows with a corpus luteum (CL) at TAI protocol commencement, carrying twins and those served during the cold season had an enhanced RIWA compared with those with no CL, carrying singletons and cows served during the hot season. In addition, healthy cows (P = 0.003) and cows showing estrus (P = 0.0002) had enhanced pregnancy per TAI compared with those with postpartum disorders, and those failing to exhibit estrus, respectively. Hence, cows that remained healthy during the postpartum had a greater RIWA, expression of behavioral estrus intensity and pregnancy to TAI compared with those that had a postpartum disorder.

Advanced diffusion-weighted imaging-derived quantitative parameters as biomarkers of fibrosarcoma-cell proliferation in nude mice: A study based on precise imaging-pathology correlation.

To determine whether quantitative parameters derived using diffusion kurtosis imaging (DKI) and intravoxel incoherent motion (IVIM) imaging reflect pathological changes in fibrosarcoma. Thirty nude mouse models of fibrosarcoma underwent T1/T2-weighted imaging, DKI, and IVIM imaging on a 3.0-T scanner. Immunohistochemistry was utilized for the hematoxylin and eosin, aquaporin 1 (AQP1), aquaporin 4 (AQP4), and Ki-67 staining of fibrosarcoma tissue, and AQP1 and AQP4 staining of normal muscle tissue (NMT). The independent-sample t-test was used to compare AQP1 and AQP4 expression in fibrosarcoma and NMT. Pearson and Spearman correlation analyses were conducted to evaluate the correlation between imaging parameters and pathological indicators. Multiple linear regression analysis was employed to identify the pathological indicators independently associated with quantitative DKI and IVIM parameters. Apparent diffusion coefficient (ADC), D, f, and mean kurtosis (MK) indicated cell density and Ki-67 and AQP1 expression intensity. D values reflected AQP4 expression intensity, while MD reflected cell density and AQP1 expression intensity. Cell density (CD) independently influenced ADC and f values, while CD and AQP1 independently influenced D values. CD and Ki-67 independently influenced MK. DKI- and IVIM imaging-derived ADC, D, f, MD, and MK were correlated with AQP1, AQP4, Ki-67, and CD in nude mice with fibrosarcoma.

Иммуноморфологическая характеристика склероатрофического лихена в раннем и позднем периоде

Introduction. Lichen sclerosus is a chronic inflammatory disease of the skin and mucous membranes, combining degenerative changes in the epidermis and dermis. Histological examination is needed due to diverse nosologies with similar clinical manifestations. Early development stages of lichen sclerosus are characterized by nonspecific histological changes. However, some changes can serve as indicators for accurate diagnosis. Materials and methods. The diagnosis was confirmed by typical clinical manifestations, medical history, and laboratory abnormalities. Biopsy samples were taken from early-stage lesions and recent rash sites. The patients were divided into two groups: group 1 consisted of patients with clinical manifestations of the disease lasting less than one year and group 2 included the patients who had symptoms for more than two years. Serial paraffin sections were stained with hematoxylin and eosin as well as Van Gieson’s picric fuchsin. We performed immunohistochemical analysis using polyclonal antibodies against CD95 and monoclonal antibodies against CD31. We evaluated the findings of the immunohistochemical assay semi-quantitatively in each case for every marker. Morphometric analysis of microvessels was conducted, and the expression intensity was determined. Results. We identified new extended criteria for the differential diagnosis of lichen sclerosus including the disease duration (the presence and prevalence of rashes, erosions, presence or absence of cracks), evaluation of the microcirculatory involvement, determination of the percentage of vascular damage, and the assessment of the severity of endothelial changes. Conclusion. By means of morphological analysis, we determined the diagnostic criteria for early and late stages of lichen sclerosus. Our research results and literature data analysis showed that damaged vascular wall was the main and determining factor of lichen sclerosus pathogenesis. Keywords: lichen sclerosus, histological analysis, immunohistochemical analysis, diagnostic criteria, vascular damage