Fibrous Areas Research Articles

Trans-rectovesical pouch urethral-sparing robotic-assisted simple prostatectomy: A case series.

To detail a novel technique of robotic-assisted simple prostatectomy that makes handling the gland protruding into the bladder neck easier and can preserve the urethra and retain ejaculation function as much as possible. This is a prospective case series. Clinical data of 17 male patients who had large volume (>80 mL) benign prostatic hyperplasia (BPH) were enrolled to undergo trans-rectovesical pouch urethral-sparing robotic-assisted simple prostatectomy (usRASP). We adopted the approach through the space between the bladder neck and seminal vesicle to perform a usRASP that can avoid the detrusor skirt and fibrous matrix area of the retropubic prostate. Between the transitional zone and the peripheral zone of the large prostate, the hyperplastic prostatic gland tissue can be enucleated under direct vision while preserving the prostatic urethra and retaining the ejaculatory duct and bladder neck intact. All preoperative, perioperative and postoperative clinical data were collected, and descriptive analysis was performed. The median intravesical prostatic protrusion was 19.3 mm (8.5-32.2). The median operative time was 100 min (75-140), and the median estimated blood loss was 100 mL (10-500). The median time to catheter removal was 7 days (5-7), with a median postoperative hospital stay of 2 days (2-4). After at least 6-month follow-up, the median maximum urine flow rate and postvoid residual volume were 40.1 mL/s (12.7-52.4) and 15 mL (5-23), respectively; the median International Prostate Symptom Score and Quality of Life score were 0 (0-6.3) and 1 (0-3), respectively; and the median total prostate-specific antigen was 0.84 ng/mL (0.15-1.01). All patients successfully underwent usRASP. Fifty-eight percent of patients with normal ejaculation function before surgery can still retain normal ejaculation function. We described a new approach to performing usRASP. This new method remarkably improved the voiding function, maintained antegrade ejaculation and did not increase the post-operative complications.

Seromucinous tumors of ovary: A case series

Seromucinous neoplasms are new category of ovarian epithelial tumor in the 2014 revised World Health Organization classification. Surface epithelial neoplasms are the most common ovarian tumors. Based on the degree of stratification and the presence or absence of invasion, these epithelial proliferations are further divided into benign, borderline, and malignant categories. Benign tumors are divided into three subtypes: cystadenoma (cystic areas), cystadenofibroma (cystic and fibrous areas), and adenofibroma (predominantly fibrous sections). Serous adenofibromas are rare variant of serous surface epithelial tumors. They affect females between the ages of 15 and 65 and make up 1.7% of all benign ovarian tumors. Serous tumor with a solid, fibrous component is designated as serous cystadenofibroma in which both serous and mucinous components are closely intermixed. Seromucinous adenofibroma is a benign cystic neoplasm composed of glands and cysts lined by an admixture of bland mullerian type epithelium which includes ciliated, endocervical type mucinous, and endometroid. They have a characteristic prominent fibromatous hypocellular stroma which helps differentiate it from the commoner cystadenoma. Seromucinous borderline tumors is 10% -20% of all epithelial ovarian tumors. Develops in younger age, usually bilateral in upto 40%, associated with endometriosis. We discuss about four different cases of seromucinous tumor reported in our institute.

Fibulin-4 as a potential extracellular vesicle marker of fibrosis in patients with cirrhosis.

Chronic liver injury leads to decreased liver function and increased fibrosis. Fibrosis is not only associated with the development of portal hypertension and carcinogenesis, but with the occurrence of events and a poor prognosis, highlighting the importance of non-invasive fibrosis assessment in patients. In the present study, we searched for markers related to liver fibrosis via proteomic analysis of small extracellular vesicles (sEVs). In the discovery cohort, proteomic analysis was carried out in the sEVs extracted from the sera of 5 patients with decompensated cirrhosis, 5 patients with compensated cirrhosis, and 5 controls without liver disease. Interestingly, in this cohort, fibulin-4 was significantly associated with cirrhosis while in the validation cohort [formed by 191 patients: 7 patients without disease, 16 patients without liver disease (other diseases), 38 patients with chronic liver disease (CLD), 75 patients with cirrhosis of Child-Pugh class A (36 without hepatocellular carcinoma [HCC], 29 with HCC), and 65 patients with cirrhosis of Child-Pugh class B-C (39 without HCC, 26 with HCC)], fibulin-4/CD9 levels increased with cirrhosis progression. Furthermore, the fibulin-4/CD9 ratio was significantly higher in patients with varices. Immunostaining also revealed strong fibulin-4 expression in cholangiocytes within the fibrous areas and mesothelial cells in liver tissue blood vessels. Taken together, our results suggest that fibulin-4, essential for lysyl oxidase activation, might be a new liver fibrosis marker found in the sEVs of patients with cirrhosis.

Impact of endothelial shear stress on coronary atherosclerotic plaque progression and composition: A meta-analysis and systematic review

Background and aimsIntracoronary pressure gradients and translesional flow patterns have been correlated with coronary plaque progression and lesion destabilization. In this study, we aimed to determine the relationship between endothelial shear stress and plaque progression and to evaluate the effect of shear forces on coronary plaque features. MethodsA systematic review was conducted in medical on-line databases. Selected were studies including human participants who underwent coronary anatomy assessment with computational fluid dynamics (CFD)-based wall shear stress (WSS) calculation at baseline with anatomical evaluation at follow-up. A total of six studies were included for data extraction and analysis. ResultsThe meta-analysis encompassed 31′385 arterial segments from 136 patients. Lower translesional WSS values were significantly associated with a reduction in lumen area (mean difference −0.88, 95% CI −1.13 to −0.62), an increase in plaque burden (mean difference 4.32, 95% CI 1.65 to 6.99), and an increase in necrotic core area (mean difference 0.02, 95% CI 0.02 to 0.03) at follow-up imaging. Elevated WSS values were associated with an increase in lumen area (mean difference 0.78, 95% CI 0.34 to 1.21) and a reduction in both fibrofatty (mean difference −0.02, 95% CI −0.03 to −0.01) and fibrous plaque areas (mean difference −0.03, 95% CI −0.03 to −0.03). ConclusionThis meta-analysis shows that WSS parameters were related to vulnerable plaque features at follow-up. These results emphasize the impact of endothelial shear forces on coronary plaque growth and composition. Future studies are warranted to evaluate the role of WSS in guiding clinical decision-making.

Immunohistochemical expression of H3.3 G34W in 100 giant cell tumors of bone and its diagnostic mimics, including its value in resolving uncommon diagnostic scenarios: A single institutional study at a tertiary cancer referral center, India.

There can be a diagnostic challenge in differentiating giant cell tumor of bone (GCTB) from its mimics. Lately, histone H 3 F 3 A (Histone 3.3 ) G34W has been identified as a promising immunohistochemical marker. This study was aimed at evaluating H3.3 G34W immunostaining in 100 GCTBs, including its value in resolving diagnostic dilemmas. Immunohistochemical staining for H3.3 G34W was graded in terms of staining intensity (1+ to 3+) and the percentage of tumor cells showing crisp nuclear staining. One hundred GCTBs occurred in 58 males and 42 females (M: F ratio = 1.3), of 7-66 years age (average = 31.3, median = 28), commonly in distal femur (26), followed by proximal tibia (17), distal radius (12), proximal humerus (7), metacarpals (7), sacrum (6), proximal fibula (6), and relatively unusual sites (19), including a single multicentric case. Out of 92 GCTBs, wherein H3.3 G34W immunostaining worked, 81 (88.1%) showed positive staining in the mononuclear cells, including tumors with fibrous histiocytoma-like areas, sparing osteoclast-like giant cells, with 3+ staining intensity in 65/81 (80%) tumors. All 7/7 (100%) malignant GCTBs showed positive staining, including the pleomorphic/sarcomatous cells. All 7/7 (100%) metastatic GCTBs showed positive immunostaining. Seven out of 10 post-denosumab treated GCTBs showed positive H3.3 G34W immunostaining in the residual mononuclear cells. None of the other 37 "giant cell-rich" lesions displayed H3.3 G34W immunostaining. Four of 9 GCTBs tested for H3.3 G34W mutation showed positive results. The diagnostic sensitivity and specificity of H3.3 G34W for GCTB were 88.1% and 100%, respectively. This constitutes one of the first reports from our country, further validating the diagnostic value of H3.3 G34W in differentiating GCTB, including metastatic and malignant forms from its mimics, including small biopsy samples. Its value in various diagnostic dilemmas is presented and utility in identifying residual tumor cells in post-denosumab treated GCTBs is worth exploring.

Evaluation of ARFI elastography for detecting active mastitis in sheep with previous fibrous lesions: a study of mammary parenchyma and supramammary lymph nodes.

The aim of the study was to evaluate the use of Acustic Radiation Force Impulse (ARFI) elastography in mammary parenchyma and supramammary lymph nodes, for detection of active mastitis in sheep with naturally infected chronic fibrous lesions. 27 female sheep were included and B-mode ultrasound and ARFI elastography images were obtained, acquiring qualitative (echogenicity and echotexture) and quantitative (shear rate, depth and short/long axis ratio) variables of 48 mammary glands. The glands were divided into three experimental groups: control group (CG) - healthy animals; LSCC- animals that presented fibrous lesions and SCC (somatic cell count) less than 500 x 103 cls/mL; HSCC: animals that presented fibrous lesions and SCC (somatic cell count) more than 500 x 103 cls/mL; The qualitative variables using B-mode ultrasonography, including echotexture and echogenicity, showed no significant differences between the evaluated groups and tissues (p = 0.9336 and p = 0.233, respectively) .In healthy areas of the gland, it was an increasing in shear wave velocity (SWV) in LSCC than in HSCC (p=0.04). When comparing the fibrosis in the LSCC and HSCC groups with their respective normal areas, the velocity increased in both groups: LSCC (p= 0,0007) and HSCC (p= 0,0001). When comparing the areas of fibrosis in LSCC and HSCC with the CG parenchyma, there was an increase in LSCC (p=0.001) and HSCC (p=0.0001). B-mode ultrasound indicate predominance of hypoechoic echogenicity in lymph nodes and reduced short/long axis ratio in cases of active subclinical mastitis. The supramammary lymph node showed increased SWV when comparing the CG with HSCC groups (p=0.02) and GC with LSCC (p=0.04). B-mode ultrasonography is useful for evaluating the mammary parenchyma, however, its application as a standalone diagnostic technique is not recommended. ARFI elastography indicates potential cutoff points for differentiating subclinical mastitis from healed mastitis, highlighting its importance as a tool for distinguishing normal areas from fibrous parenchymal areas. While this study did not establish specific cutoff points due to sample size limitations, further research with larger sample sizes could explore and define these critical thresholds.

The effect of BMSCs on implantation mechanisms and pregnancy process in the experimental Asherman model.

In this study, we aimed to investigate the role of bone marrow-derived mesenchymal stem cells (BMSC) on the molecular mechanisms involved in the removal of endometrial adhesions and the implantation process in the experimentally-created Asherman syndrome model in rats. An experimental model chemical agent was used. Culture medium (CM), only BMSC, 48-hour culture medium (Niche), and BMSC+ niche were used as therapy. Each group was divided into two subgroups, and changes in the endometrial tissue were investigated by histochemical and immunohistochemical staining. The results of the study show that BMSCs exhibit therapeutic properties in endometrial damage, increase endometrial thickness and vascularization, and contribute to the implantation of embryos by reducing fibrous areas. These results are promising for the prevention of Asherman-induced infertility. It has been shown that BMSC and Niche applications can be helpful in preventing adhesion molecules in the mechanisms that cause infertility.

Integrin αV mediated activation of myofibroblast via mechanoparacrine of transforming growth factor β1 in promoting fibrous scar formation after myocardial infarction

BackgroundMyocardial infarction (MI) dramatically changes the mechanical stress, which is intensified by the fibrotic remodeling. Integrins, especially the αV subunit, mediate mechanical signal and mechanoparacrine of transforming growth factor β1 (TGF-β1) in various organ fibrosis by activating CFs into myofibroblasts (MFBs). We investigated a possible role of integrin αV mediated mechanoparacrine of TGF-β1 in MFBs activation for fibrous reparation in mice with MI. MethodsHeart samples from MI, sham, or MI plus cilengitide (14 mg/kg, specific integrin αV inhibitor) treated mice, underwent functional and morphological assessments by echocardiography, and histochemistry on 7, 14 and 28 days post-surgery. The mechanical and ultrastructural changes of the fibrous scar were further evaluated by atomic mechanics microscope (AFM), immunofluorescence, second harmonic generation (SHG) imaging, polarized light and scanning electron microscope, respectively. Hydroxyproline assay was used for total collagen content, and western blot for protein expression profile examination. Fibroblast bioactivities, including cell shape, number, Smad2/3 signal and expression of extracellular matrix (ECM) related proteins, were further evaluated by microscopic observation and immunofluorescence in polyacrylamide (PA) hydrogel with adjustable stiffness, which was re-explored in fibroblast cultured on stiff matrix after silencing of integrin αV. The content of total and free TGF-β1 was tested by enzyme-linked immunosorbent assay (ELISA) in both infarcted tissue and cell samples. ResultIncreased stiffness with heterogeneity synchronized with integrin αV and alpha smooth muscle actin (α-SMA) positive MFBs accumulation in those less mature fibrous areas. Cilengitide abruptly reduced collagen content and disrupted collagen alignment, which also decreased TGF-β1 bioavailability, Smad2/3 phosphorylation, and α-SMA expression in the fibrous area. Accordingly, fibroblast on stiff but not soft matrix exhibited obvious MFB phenotype, as evidenced by enlarged cell, hyperproliferation, well-developed α-SMA fibers, and elevated ECM related proteins, while silencing of integrin αV almost abolished this switch via attenuating paracrine of TGF-β1 and nuclear translocation of Smad2/3. ConclusionThis study illustrated that increased tissue stiffness activates CFs into MFBs by integrin αV mediated mechanoparacrine of TGF-β1, especially in immature scar area, which ultimately promotes fibrous scar maturation.

Selective inhibition of hepatic stellate cell and fibroblast-derived LOXL1 attenuates BDL- and Mdr2-/--induced cholestatic liver fibrosis.

Lysyl oxidase-like 1 (LOXL1) proteins are amine oxidases that play a crucial role in extracellular matrix remodeling due to their collagen cross-linking and intracellular functions. The role of LOXL1 in cholestatic liver fibrosis remains unexplored. We measured LOXL1 expression in two murine models of cholestasis [Mdr2 knockout (Mdr2-/-) and bile duct ligation (BDL)]. We used adeno-associated virus (AAV) serotype 6-mediated hepatic delivery against LOXL1 (AAV2/6-shLoxl1) to investigate the therapeutic efficacy of targeting LOXL1 in cholestatic liver fibrosis. NIH-3T3 murine fibroblasts were used to investigate the function and regulatory mechanisms of LOXL1 in vitro. LOXL1 expression was significantly upregulated in Mdr2-/- and BDL mice compared with their corresponding controls, predominantly in collagen-rich fibrous septa and portal areas. AAV2/6-shLoxl1 significantly reduced LOXL1 levels in Mdr2-/- and BDL mice, mainly in desmin-positive hepatic stellate cells (HSCs) and fibroblasts. Concomitant with reduced LOXL1 expression, there was reduced ductular reaction, inflammation, and fibrosis in both Mdr2-/- and BDL mice. In addition, Loxl1 intervention decreased Ki-67-positive cells in the desmin-positive areas in both Mdr2-/- and BDL mice. Overexpression of LOXL1 significantly promoted fibroblast proliferation by activating the platelet-derived growth factor receptor and extracellular signal-regulated kinase signaling pathways in vitro. Our findings demonstrated that selective inhibition of LOXL1 derived from HSCs/fibroblasts attenuated cholestatic liver/biliary fibrosis, inflammation, ductal reaction, and HSC/fibroblast proliferation. Based on our findings, LOXL1 could be a potential therapeutic target for cholestatic fibrosis.NEW & NOTEWORTHY Selectively, inhibition of HSC/fibroblasts-derived LOXL1 by AAV2/6-shLoxl1 could reduce collagen deposition, HSC/fibroblasts proliferation, and cholestatic liver fibrosis progression. In addition, overexpression of LOXL1 significantly promoted HSC/fibroblast proliferation by activating the PDGFRß/PI3K and ERK signaling pathways in vitro.

Assessment of vasa vasorum on coronary plaques in patients with acute coronary syndromes using intravascular ultrasound and iMap analysis: A retrospective cohort study.

Studies have revealed that vasa vasorum (VV) neovascularization is vital for the progression and vulnerability of coronary atherosclerotic plaques. The correlation between VV, plaque constituents, and the no-reflow phenomenon (NRP) in percutaneous coronary intervention (PCI) remains elusive. We explored plaque constituents in iMap-intravascular ultrasound (iMap-IVUS) and NRP during PCI for VV lesions. We studied 166 coronary lesions in 166 patients with acute coronary syndromes (ACS) (118 lesions with VV) undergoing pre-intervention intravascular ultrasound (IVUS). We evaluated the diversity in plaque morphological status and post-PCI results based on the presence or absence of VV. The lesions with VV group had significantly higher high-sensitivity C-reactive protein (hs-CRP) levels than the lesions without VV group (8.41 ± 4.98 vs 4.19 ± 3.69 mg/L, P < .001). The frequency of after-stent deployment thrombolysis in myocardial infarction (TIMI) flow grades 0, 1, and 2 was remarkably greater in lesions with VV than in those without VV (22.9% vs 10.4%, P < .001). Plaques at the minimum lumen, necrotic core (1.26 ± 0.64 vs 0.92 ± 0.61 mm2, P < .001; 20.95 ± 7.19 vs 13.34% ± 6.54%, P < .001), and fibrous areas (4.23 ± 1.32 vs 3.92 ± 1.01 mm2, P = .006; 61.01 ± 9.41 vs 56.92% ± 11.42%, P = .001) were considerably larger in the lesions with VV than in those without VV. In addition, densely calcified plaques (0.41 ± 0.26 vs 0.81 ± 0.59 mm2, P < .001; 3.63 ± 2.19 vs 7.18% ± 2.01%, P < .001) were considerably smaller in the lesions with VV than in those without VV. Multivariate analyses revealed that VV and plaque volume were independent predictors of NRP after stent deployment (odds ratio [OR]: 5.13, 95% confidence interval [CI]: 1.19-15.32, P = .002; OR: 4.79, 95% CI: 1.08-9.01, P = .005). Lesions with VV exhibited considerable plaque vulnerability in patients with ACS, and they displayed more NRP during PCI. VV and plaque volume were independent predictors of NRP after stent deployment.

Neutrophil-specific STAT4 deficiency attenuates atherosclerotic burden and improves plaque stability via reduction in neutrophil activation and recruitment into aortas of Ldlr-/- mice.

Neutrophils drive atheroprogression and directly contribute to plaque instability. We recently identified signal transducer and activator of transcription 4 (STAT4) as a critical component for bacterial host defense in neutrophils. The STAT4-dependent functions of neutrophils in atherogenesis are unknown. Therefore, we investigated a contributory role of STAT4 in neutrophils during advanced atherosclerosis. We generated myeloid-specific Stat4ΔLysMLdlr-/-, neutrophil-specific Stat4ΔS100A8Ldlr-/-, and control Stat4fl/flLdlr-/- mice. All groups were fed a high-fat/cholesterol diet (HFD-C) for 28 weeks to establish advanced atherosclerosis. Aortic root plaque burden and stability were assessed histologically by Movat pentachrome staining. Nanostring gene expression analysis was performed on isolated blood neutrophils. Flow cytometry was utilized to analyze hematopoiesis and blood neutrophil activation. In vivo homing of neutrophils to atherosclerotic plaques was performed by adoptively transferring prelabeled Stat4ΔLysMLdlr-/- and Stat4fl/flLdlr-/- bone marrow cells into aged atherosclerotic Apoe-/- mice and detected by flow cytometry. STAT4 deficiency in both myeloid-specific and neutrophil-specific mice provided similar reductions in aortic root plaque burden and improvements in plaque stability via reduction in necrotic core size, improved fibrous cap area, and increased vascular smooth muscle cell content within the fibrous cap. Myeloid-specific STAT4 deficiency resulted in decreased circulating neutrophils via reduced production of granulocyte-monocyte progenitors in the bone marrow. Neutrophil activation was dampened in HFD-C fed Stat4ΔLysMLdlr-/- mice via reduced mitochondrial superoxide production, attenuated surface expression of degranulation marker CD63, and reduced frequency of neutrophil-platelet aggregates. Myeloid-specific STAT4 deficiency diminished expression of chemokine receptors CCR1 and CCR2 and impaired in vivo neutrophil trafficking to atherosclerotic aorta. Our work indicates a pro-atherogenic role for STAT4-dependent neutrophil activation and how it contributes to multiple factors of plaque instability during advanced atherosclerosis in mice.

Evolution of atrial remodeling and atrial arrhythmogenicity in a rat model of chronic pulmonary artery constriction: A “trialogue” between right heart disease, cardiac inflammation, and atrial fibrillation inducibility

Among all cardiac arrhythmias, atrial fibrillation (AF) is the most commonly diagnosed. Aging and comorbid pathological conditions such as obesity, sleep apnea or myocardial infarction increase the prevalence and incidence of AF. Clinical and experimental evidence have shown that pulmonary hypertension (PH) can initiate the development of right heart disease (RHD), which is also considered an important AF risk factor. Inflammation emerges as a common denominator between cardiovascular comorbidities and AF susceptibility. The underlying mechanisms linking (a) RHD, (b) atrial inflammation, and (c) the development of AF, remain unclear. We hypothesize that a constriction of the pulmonary artery trunk (PAT) progressively generates atrial inflammation and fibrosis responsible for increasing AF susceptibility. Describe the evolution of atrial inflammation and fibrosis in a rat model of RHD. RHD was induced on anesthetized Wistar rats (225–275 g) by performing a Pulmonary Artery Banding (PAB) which permanently reduced the PAT diameter to 1 mm. Sham animals had a PAB-mimicking surgery without suture. At day 0, 7, 14 and 21 post-surgery echocardiography and electrophysiological studies were performed in vivo on every animal. Optical mapping, histological study using Masson's trichrome, qPCR and immunoblotting were performed to analyze atrial tissues at each timepoint. PAB rats showed myocardial hypertrophy and cavity dilation affecting the right atrium (RA) and right ventricle (RV). Optical mapping performed on the RA showed that the conduction velocity was slower, the effective refractory periods were shorter, and the action potential durations were smaller in PAB rats compared to Sham. These electrical and structural remodeling were associated with significantly increased AF inducibility in PAB rats starting at day 7 until day 21 post-surgery. RA Fibrous area was also significantly higher in PAB rats compared to Sham at day 14 to day 21 post-PAT ligation. Inflammation and fibrosis-related genes such as IL6, TGFβ1 and CO1LA1 were progressively and significantly increased in the RA from PAB rats compared to Sham, from the 7th to 21st day post-PAT constriction. PAB-induced RHD provoked a progressively aggravating right atrial inflammatory and fibrotic profile leading to an increased risk of AF in rats.

Inflammatory response to epoxy resin and calcium silicate sealers preheated with different temperatures: an in vivo study.

To determine the impact of heat exposure of different sealers on inflammatory cytokine secretions and tissue response in vivo. Silicone tubes were prefilled with epoxy resin (ER) or calcium silicate (CS) sealers, preheated at 37, 60, or 120 °C, and implanted in rat subcutaneous site. Peri-implant exudate and tissue were analyzed after 1 and 4 weeks for cytokine secretions and tissue organization. At 1 week, 120 °C-preheated CS and ER induced higher secretions of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), respectively, as compared to sham/empty tube groups. At 4 weeks, whereas TNF-α secretion was reduced in CS, it increased in ER group, particularly for 120 °C. Both sealers revealed high IL-6 after 4 weeks as compared to sham/empty tube, and generally, higher IL-6 secretions were associated with ER. Histology at 1 week revealed lower degree of inflammatory infiltrate in the groups of the highest preheating temperature (120 °C). Nonetheless, at 4 weeks, whereas fibrous capsule area and inflammatory infiltrate remained low in the CS120 group, they were high in ER120. Preheating ER sealer to 120 °C induced high and prolonged secretion of proinflammatory cytokines (TNF-α and IL-6), whereas this effect was transient for the CS sealer. This was associated with increased fibrous capsule and inflammatory infiltrate in response to 120 °C-preheated ER. Heat-induced changes in sealer properties alter the inflammatory response in vivo, which may affect the clinical outcome. This will not only help appropriate selection of obturation technique for different sealers, but also for optimizing the properties of new generation of sealers.

The Role of Prostate Zones in Cancer Progression

Purpose: The prostate is composed of four main areas: anterior fibrous septal area, transition zone (TZ), peripheral zone (PZ) and central zone (CZ), with more than 95% of tumours located in PZ and TZ [1,2]. Each zone has variable tumour incidence, with patients diagnosed with TZ tumours having a better prognosis than PZ and CZ tumours [3,4]. Recent molecular and histopathological studies have shown that unique zonal features may contribute to the diverse patient outcome carrying tumours arising from each prostate zone [3–5]. This study aims to understand the precise cellular composition of the prostate zones and their functional contribution to tumour progression. We first aim to define the cellular composition of the prostate zones, focusing on the epithelial tumour cells and their microenvironment.

A case of FN1-fused calcified chondroid mesenchymal neoplasm of the hand with novel FGFR3 partner gene.

Calcified chondroid neoplasms with FN1::FGFR1 or FGFR2 fusions constitute a recently described category of mesenchymal neoplasms mostly encountered in the extremities and temporomandibular joint. Herein, we report a case of FNI1-fused calcified chondroid neoplasm of the hand with a novel FGFR3 fusion partner. The tumor exhibited a multilobulated growth pattern composed of epithelioid cells embedded in abundant stroma with myxoid, chondroid, and fibrous areas and scattered osteoclast-like giant cells. RNA sequencing revealed an in-frame fusion between Exon 31 of FN1 and Exon 3 of FGFR3, which was subsequently confirmed by reverse transcription-polymerase chain reaction. Our findings expand on the spectrum of potential fusion partners in FN1-fused calcified chondroid neoplasms.

Focal Nodular Hyperplasia-Like Lesions With Glypican-3 Positivity in Infancy.

Glypican-3 is often used to discriminate between neoplastic and nonneoplastic liver. In focal lesions, positivity may be considered suggestive of a malignancy such as hepatoblastoma. However, glypican-3 is also normally expressed in the immature liver. We present a series of 5 cases of focal nodular hyperplasia (FNH)-like lesions arising in very young patients with glypican-3 expression and highlight the challenges these lesions present in the differential diagnosis of hepatoblastoma. Cases were obtained from the files of 3 tertiary pediatric hospitals. Clinical data were obtained from the electronic medical record and histopathologic material including immunohistochemical stains were reviewed. Patients were aged 2 weeks to 6 months with peak AFP levels ranging from 88.6 to 204,696 ng/mL. Microscopically, all were variably demarcated hepatocellular lesions with cords of hepatocytes, marked sinusoidal dilatation, and occasional fibrous bands and areas reminiscent of central scar with bile ducts. No significant cytologic atypia or increased mitotic activity were present. All showed glypican-3 expression and were negative for nuclear beta-catenin with intact reticulin framework. Our study highlights the pitfalls of evaluating focal liver lesions in infants when high AFP levels and glypican-3 expression may reflect immaturity rather than neoplasia.

Brain White Matter Structural Alteration in Hemifacial Spasm: A Diffusion Tensor Imaging Study

To analyze the changes in the white matter structure of the whole brain in hemifacial spasm (HFS) patients by using the tract-based spatial statistics (TBSS) method. 29 HFS patients without anxiety and depression and 29 healthy controls with matching age, sex, and education were selected. All subjects received a 3.0T magnetic resonance (MR) brain diffusion tensor imaging scan. Tract-based spatial statistics method was used to analyze the changes in white matter structure in the whole brain and obtained the cerebral white matter fibrous areas exhibiting significant intergroup differences. The fractional anisotropy (FA), mean diffusivity, axial diffusivity, and radial diffusivity of these areas were abstracted. Analyzed the correlation between these diffusion metrics and clinical variables (disease duration, spasm severity). Compared with the healthy controls group, the HFS group exhibited significantly lower FA in the forceps minor, bilateral anterior thalamic radiation, and right superior longitudinal fasciculus ( P <0.05, threshold-free cluster enhancement corrected). Cohen grading scale of HFS patients was negatively correlated with FA of forceps minor. Based on TBSS analysis, the injury of white matter fiber tracts in HFS patients was found, including forceps minor, bilateral anterior thalamic radiation, and right superior longitudinal fasciculus. The changes of FA values in forceps minor were negatively correlated with the Cohen grading scale, suggesting that the alteration of white matter fiber in the genu-of-corpus-callosum-cortex circuit plays an important role in the neuro-pathological mechanism of HFS. Combined with previous research, it is also necessary to further explore the change of the superior longitudinal fasciculus in the future.

Abstract 13103: Defective Vascular Smooth Muscle Cell Tafazzin Impairs Mitochondrial Function and Promotes Atherosclerosis

Introduction: Tafazzin acylates immature monolysocardiolipin to form mature cardiolipin. Cardiolipin is the signature phospholipid of the mitochondrial inner membrane required for respiratory efficiency and regulation of apoptosis. Mitochondrial respiration is reduced in atherosclerosis. However, whether tafazzin regulates vascular smooth mucle cell (VSMC) mitochondrial function and contributes to atherosclerosis is unclear. Hypothesis: Tafazzin regulates VSMC mitochondrial function and atherosclerosis development. Methods: We examined tafazzin expression and regulation in human plaques and VSMCs. We determined the in vitro effects of tafazzin using siRNA mediated cell silencing or lentiviral infection to express either tafazzin (Taz) or tafazzin with a transacylase defective mutation (Taz H69Q ). We also generated mice expressing VSMC restricted Taz or Taz H69Q and crossed them with apolipoprotein E deficient mice (SM22α-Taz/ApoE -/- or SM22α-Taz H69Q /ApoE -/- ) to study tafazzin in vivo Results: Tafazzin mRNA and protein is decreased in plaque and plaque VSMCs. MicroRNA 125a-5p expression is increased in plaques, downregulates tafazzin expression and is induced by oxidised low density lipoprotein. Tafazzin silencing and expression of Taz H69Q reduces mitochondrial respiration, decreases ATP content (0.13±0.01nmoles vs 0.21±0.01nmole, n=4), and promotes apoptosis. SM22α-Taz H69Q /ApoE -/- mice showed increased plaque burden (28±2.3% vs 17±2.1%, n=17), increased necrotic core and reduced fibrous cap areas. In contrast, over-expression of tafazzin increases VSMC mitochondrial respiration and protects against apoptosis. SM22α-Taz/ApoE -/- mice showed no change in plaque burden but increased features of plaque stability. SS-31, which binds and stabilises cardiolipin, improves mitochondrial respiration and decreases apoptosis in SM22α-Taz H69Q /ApoE -/- VSMCs. Conclusions: Tafazzin expression is decreased in plaque VSMCs and is negatively regulated by microRNA 125a-5p. Defective tafazzin decreases mitochondrial respiration, increases apoptosis and promotes atherosclerosis. Tafazzin is therefore an important and unrecognised regulator of VSMC mitochondrial function and atherosclerosis.

Abstract P3032: Liver-directed Mitochondrial Uncoupling Attenuates The Progression Of Early And Late-stage Atherosclerosis In Mice

Atherosclerotic cardiovascular disease (CVD) remains a leading cause of morbidity and mortality in people with type 2 diabetes (T2D) and new therapies are urgently needed. We recently developed an orally available, liver-directed controlled-release mitochondrial protonophore (CRMP) and showed that this agent safely reverses hypertriglyceridemia, fatty liver, and T2D in rodent and nonhuman primate models of obesity by promoting a subtle sustained increase in hepatic mitochondrial inefficiency. To determine the effect of CRMP on atherogenesis, we performed intervention studies in which 12-week Western-diet fed Ldlr -/- mice with established atherosclerosis were treated with CRMP (30 mg/kg-day) or vehicle control for an additional 12 weeks. CRMP-treated mice displayed a significant reduction in fasting plasma and hepatic triglyceride content, independently of changes in body weight, fasting plasma glucose, insulin or cholesterol levels. Morphometric analysis of the aortic root and brachiocephalic artery revealed that CRMP treatment significantly reduced total plaque area and neutral lipid accumulation. CRMP treatment also decreased markers of plaque vulnerability, as evidenced by a reduction in necrotic core area and increase in fibrous cap area ( P &lt;0.05 vs veh). To determine the mechanisms by which CRMP attenuated atherosclerosis, we performed progression studies in Ldlr -/- mice fed a Western diet containing CRMP or vehicle control for 12 weeks. Similarly to the intervention studies, CRMP significantly reduced aortic root plaque area, neutral lipid accumulation and necrotic core area. These changes were associated with a reduction in plasma triglyceride content and improvement in whole-body insulin sensitivity as assessed by hyperinsulinemic-euglycemic clamps (all P &lt;0.05 vs veh). Single-cell RNA sequencing of whole aortas from CRMP and vehicle-treated Ldlr -/- mice revealed that CRMP reduced the frequency of inflammatory macrophages compared to vehicle (~19% vs ~2%), consistent with an immune-dampening effect of CRMP. Taken together, these data support an anti-atherogenic role for CRMP and suggest the therapeutic potential of liver-directed mitochondrial uncouplers for the treatment of early and late-stage atherosclerosis.

Validation of the modified radiographic union score for tibia fractures (mRUST) in murine femoral fractures.

Bony union is a primary predictor of outcome after surgical fixation of long bone fractures. Murine models offer many advantages in assessing bony healing due to their low costs and small size. However, current fracture recovery investigations in mice frequently rely on animal sacrifice and costly analyses. The modified Radiographic Union Score for Tibia fractures (mRUST) scoring system is a validated metric for evaluating bony healing in humans utilizing plain radiographs, which are relatively inexpensive and do not require animal sacrifice. However, its use has not been well established in murine models. The aim of this study was to characterize the longitudinal course of mRUST and compare mRUST to other conventional murine fracture analyses. 158 mice underwent surgically created midshaft femur fractures. Mice were evaluated after fracture creation and at 7, 10, 14, 17, 21, 24, 28, 35, and 42 days post-injury. mRUST scoring of plain radiographs was performed by three orthopaedic surgeons in a randomized, blinded fashion. Interrater correlations were calculated. Micro-computed tomography (μCT) was analyzed for tissue mineral density (TMD), total callus volume (TV), bone volume (BV), trabecular thickness, trabecular number, and trabecular separation. Histomorphometry measures of total callus area, cartilage area, fibrous tissue area, and bone area were performed in a blinded fashion. Ultimate torque, stiffness, toughness, and twist to failure were calculated from torque-twist curves. A sigmoidal log-logistic curve fit was generated for mRUST scores over time which shows mRUST scores of 4 to 6 at 7 days post-injury that improve to plateaus of 14 to 16 by 24 days post-injury. mRUST interrater correlations at each timepoint ranged from 0.51 to 0.86, indicating substantial agreement. mRUST scores correlated well with biomechanical, histomorphometry, and μCT parameters, such as ultimate torque (r=0.46, p<0.0001), manual stiffness (r=0.51, p<0.0001), bone percentage based on histomorphometry (r=0.86, p<0.0001), cartilage percentage (r=-0.87, p<0.0001), tissue mineral density (r=0.83, p<0.0001), BV/TV based on μCT (r=0.65, p<0.0001), and trabecular thickness (r=0.78, p<0.0001), among others. These data demonstrate that mRUST is reliable, trends temporally, and correlates to standard measures of murine fracture healing. Compared to other measures, mRUST is more cost-effective and non-terminal. The mRUST log-logistic curve could be used to characterize differences in fracture healing trajectory between experimental groups, enabling high-throughput analysis.