Anti-smooth Muscle Actin Antibody Research Articles

The Predictive Value of Autoantibodies in Determining Autoimmune Hepatitis (AIH) Severity

Background: The presence of autoantibodies is a prerequisite for the diagnosis of autoimmune hepatitis (AIH). However, most autoantibodies are not disease-specific, and serological overlap between AIH and other chronic liver diseases is common. Since the prognostic parameters of AIH are limited, this study aimed to investigate the relationship between histopathological findings on liver biopsy with different types of autoantibodies associated with AIH and how autoantibodies can predict the severity and extent of disease. Methods: The present study was performed on 30 patients with a definite diagnosis of AIH according to the International Autoimmune Hepatitis Group (IAIHG) criteria. Pediatric AIH patients underwent liver tissue examinations at the time of diagnosis at accession, which confirmed characteristic histological changes. AIH-related serologic major and minor autoantibodies were measured using indirect immunofluorescence assays and ELISA kit (EUROIMMUN, Germany), respectively, and were compared within all patients, and the results were recorded. Finally, the obtained data were analyzed using SPSS V25 software. Results: Out of 30 patients, 17 (56.66%) were female, and the age range of patients was 17-11 years (8.46±6.95). Anti-nuclear antibody (ANA) (73.3%), smooth muscle antibody (SMA)-anti-smooth muscle actin antibody (ASMA) (70%), perinuclear anti-neutrophilic cytoplasmic antibodies (p-ANCA) (63%), and liver kidney microsomal (LKM) (43.3%) were the most common autoantibodies found in children with AIH. There was a significant relation between the severity of histological findings and the presence of LKM antibodies (P<0.05). The highest sensitivity for predicting severe AIH based on histopathological findings was ANA autoantibody positivity and the presence of at least two primary autoantibodies (LKM and SMA-ASMA). On the other hand, positive LKM antibodies had the highest specificity and positive predictive value (PPV) in AIH severity prediction. Conclusion: The results of the present study suggested that there might be a significant correlation between the presence of primary LKM autoantibodies and biopsy results, so it can possibly act as an accurate autoantibody for predicting the severity of AIH, while other AIH-related autoantibodies did not seem to have a significant correlation with biochemical and histological findings.

Anatomical and Histological Study of the Upper Tarsus in Asian.

The aim of this study was to observe and analyze the anatomical and histological characteristics of the upper tarsus in Asian. A total of 15 Asians (14 adults, 1 child) were used. The sagittal sections with thickness of 3μm in the middle of the upper eyelid were prepared and stained with hematoxylin-eosin, Masson trichrome and anti-smooth muscle actin antibody staining, and then were observed and photographed under light microscopy. On the sagittal section, the upper end of the tarsus was connected with the Müller muscle and the part of the posterior layer of the levator aponeurosis; the lower end was the inflated part of the palpebral margin; the superficial surface was connected with the pretarsal fascia; and the deep surface was connected with Müller muscle aponeurosis and palpebral conjunctiva. Histologically, the tarsus was similar to the structure between dense fibrous connective tissue and cartilage, and its main structures include meibomian glands and its ducts, accessory lacrimal glands, glands of Moll, glands of Zeis, eyelash hair follicles, Riolan muscle, blood vessels, and collagen fibers. Through the observation of the tarsus of a child, compared with the adult specimen, the meibomian gland tissue was more abundant; the collagen fibers density was significantly lower and arranged orderly; the blood vessel density was significantly higher; α-SMA positive smooth muscle cells could be seen in the upper end of the tarsus and its superficial and deep surfaces. In this study, the internal and adjacent anatomical structures of the upper tarsus in Asians were observed under a microscope, and according to the histological characteristics, the tarsus was divided into 3 parts and 3 types. Through the preliminary analysis, the tarsus was likely to show different histological characteristics in different individuals and age groups of Asians. There was a certain correlation between the degree of meibomian gland atrophy, the degree of collagen fibrosis and the density of blood vessel in the tarsus.

Tissue engineering of mouse uterus using menstrual blood stem cells (MenSCs) and decellularized uterine scaffold

BackgroundUterine tissue engineering can provide the opportunity for curing female infertility. Natural scaffold is a good choice to recapitulate the architecture and functionality of the native tissue. In this study, we purposed the potential of uterine decellularized scaffolds as an adequate natural niche for MenSCs differentiation toward uterus-specific cell lineages.MethodsMouse’s uterus was decellularized by immersion of hypo and hypertonic salts or freeze–thaw cycle followed by immersion in Triton X-100 and SDS solutions. MenSCs were isolated from the menstrual blood of 6 healthy women. The decellularized and recellularized samples were prepared for further in vitro and in vivo analyses.ResultsHistochemical studies and Raman spectroscopy revealed uterine ECM was preserved well, and the cells were completely removed after decellularization. Scanning electron microscopy (SEM) showed that the 3D ultrastructure of the uterus remained intact. Flowcytometric examination with CD34, CD44, CD105, CD106, CD144 markers revealed stem cell characters of cells that isolated from menstrual blood. MTT assay confirmed the bioavailability of MenSCs cultured scaffolds after 7 and 10 days.ConclusionHistochemical studies, SEM images, and Raman spectra showed MenSCs seeded and growth in uterine scaffolds. Immunostaining using anti-cytokeratin (CK), anti-desmin (Des), anti-vimentin (Vim), and anti-smooth muscle actin (SMA) antibodies showed MenSCs differentiation to epithelial and smooth muscle tissues. The Raman spectroscopy revealed the extracellular matrix (ECM) of decellularized uterine scaffolds was well preserved. The decellularized uterus can be considered a promising vehicle to support cell transplantation and differentiation. MenSCs are a good choice for uterine tissue engineering. The complete decellularization from mice uterine tissue was done by combining chemical agents

The Anatomic Features of Müller's Muscle: A Histology Study in Chinese.

Background:The purpose of this article is to clarify the character of Müller’s muscle in Chinese specimens.Methods:Ten upper eyelids of 10 formalin-fixed Chinese cadavers (9 elderly people, from 68 to 86 years of age; 1 male child, 10 years old) were examined. Full-thickness sagittal sections of the central part of the upper eyelids were microscopically examined using hematoxylin-eosin, Masson trichrome, and anti–smooth muscle actin antibodies staining.Result:In 9 elderly specimens, Müller’s muscle inserted onto tarsus via fibers, and the smooth muscle components decreased gradually and even faded away from the originated inferior branch of levator palpebrae superioris muscle to the upper margin of tarsus, although, in 1 child specimen, Müller’s muscle inserted directly onto tarsus rather than connective fibrous structure, and smooth muscle components did not decrease obviously. Both in elderly and young specimens, the aponeurotic structure that originated from Müller’s muscle extended to the tarsal plate.Conclusion:This study suggests that Müller’s muscle inserts onto the tarsus via fibrous tissue or smooth muscle, and also indicates that the smooth components of Müller’s muscle may decrease gradually in elderly people.

Abstract PD5-09: Mouse-intraductal (MIND): The first in vivo model to recapitulate the full spectrum of human DCIS pathology

Abstract Introduction. Due to advances in imaging technology and an increase in mammographic screening, there has been a significant increase in the diagnosis rate of ductal carcinoma in situ (DCIS). At the present time, nearly all women undergo surgical removal of DCIS, often followed by adjuvant radiation and in some cases anti-hormonal therapy. Currently, there are no means by which to diagnose DCIS accurately, or to predict which patients benefit from aggressive therapy. Thus, the recommendation for surgery persists, despite studies which support that not all DCIS will subsequently progress to invasive disease. In this context, animal models can be particularly useful in studying DCIS progression. Here, we present the first in vivo model of DCIS, referred to as Mouse-INtraDuctal (MIND), in which patient-derived DCIS epithelial cells are injected intraductally and allowed to progress naturally in mice. Methods. We performed intraductal injection of DCIS epithelial cells derived from 30 patient samples into 194 total glands. Of the 194, 146 xenografts showed in vivo growth, for a 75% take rate). Among the DCIS samples injected into mice, 18 (103 mouse mammary glands) were followed for a median of 9 months . Among those, 50% (9) showed invasive progression while 50% (9) remained non-invasive. DCIS invasive progression was evaluated by performing immunofluorescene staining using anti-smooth muscle actin (SMA) antibody and confirmed by the loss of SMA around the xenografted DCIS like lesions on 3 consequetive sections of FFPE tissues. Results. Progressed xenografts exhibited invasive progression, evident by the loss of SMA, as early as 6 months following transplantation. Similar to human DCIS, the cancer cells initially formed in situ lesions inside the mouse mammary ducts and mimicked all histologic subtypes including micropapillary, papillary, cribriform, solid and comedo. Among the biomarkers tested, including ER, PR, Ki67, HER2, p53, histology, nuclear and tumor grade, only low ER & PR expression and extent of DCIS growth in xenografts significantly correlated with invasive progression. A high depth targeted sequencing platform (T200) on DNA isolated from LCM captured DCIS of patient and xenograft pairs identified shared (i.e., EGFR) as well as unique (STK11, RUNX1, PIK3CA) mutations in patient/xenograft pairs. Notably, we also observed private mutations that were not shared within the same patient/xenograft pairs. These results indicate the presence of DCIS clonal heterogeneity and that DCIS xenografts may represent one or more clonal subpopulations of patient DCIS. Conclusion. The MIND model represents the first realistic in vivo model that recapitulates human DCIS progression in a manner that represents the inter- and intra-tumoral heterogeneity of human disease. These innovative mouse models will be invaluable for the discovery of molecular signatures of invasive DCIS by allowing comparison of xenografts with variable propensity for invasive progression. These models will enable the discovery of extrinsic factors that regulate DCIS malignancy as well as testing of pharmaceutical and natural compounds for prevention of DCIS progression to invasive disease. Citation Format: Fariba Behbod, Yan Hong, Darlene Limback, Hannan S Elsarraj, Haleigh Harper, Haley Haines, Hayley Hansford, Michael Ricci, Carolyn Kaufman, Mingchu Xu, Jianhua Zhang, Lisa May, Therese Cusick, Marc inciardi, Mark Redick, Jason Gatewood, Alison Aripoli, Ashley Huppe, Onalisa Winblad, Christa Balanoff, Jamie Wagner, Amanda L Amin, Kelsey E. Larson, Lawrence Ricci, Ossama Tawfik, Hana Razek, Ruby O Meierotto, Rashna Madan, Andrew K Godwin, Jeffrey Thompson, Andy Futreal, Alastair Thompson, Shelley Hwang, Fang Fan, On behalf of the Grand Challenge PRECISION consortium. Mouse-intraductal (MIND): The first in vivo model to recapitulate the full spectrum of human DCIS pathology [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PD5-09.

A New Anatomy Finding of Levator Aponeurosis and Müller Muscle at Pretarsal Plate in Asian Males.

To examine the fine anatomic structures between levator aponeurosis and Müller muscle in front of the tarsus. Postmortem specimens of 6 Chinese males (5 elderly men, aging from 68 to 86 years; 1 child, 10 years old) were used. A 3-μm thickness sagittal section of the central part of the upper eyelid was prepared, and the samples were examined microscopically by using hematoxylin-eosin, Masson trichrome, and anti-smooth muscle actin antibodies staining. There are 2 new findings in this study, one is the posterior layer of the levator aponeurosis and the other is the extensions of Müller muscle. The posterior levator aponeurosis had different insertion patterns that approximately paralleled the extension line of the levator aponeurosis at the confluence of attachment site of the orbital septum on the levator superioris. Below the confluence, it took the form of a layered insertion, and then extended to the orbicularis oculi muscle and subcutaneous tissue rather than inserting directly into tarsus. The Müller muscle was a multilayered structure at the upper border of tarsus. The superior Müller muscle extended above the peripheral arcade, and the inferior Müller muscle tendon was attached to the surface of tarsus with an Umbrella-shaped fiber. The authors discovered that the levator aponeurosis had different insertion patterns of the posterior layers. The Müller muscle gave branches at the peripheral arcade: the anterior one crossed the peripheral arcade and extended to the tarsus, and the posterior one attached and extended to the tarsus.

Leiomiosarcoma peritoneal en un canino: reporte de caso

El propósito de este estudio fue para presentar y documentar un leiomiosarcoma peritoneal en un canino y sus hallazgos histopatológicos e Inmunohistoquímica. El paciente, una hembra de 5 años de edad, de raza Chow-Chow, fue llevada a la Clínica Veterinaria de la Universidad de los Llanos, debido a pérdida de peso, postración y baja condición corporal. Al examen físico presentó mucosas pálidas, aumento de la silueta abdominal con contenido de fluido serosanguinolento. A la palpación se evidenció en el epigastrio craneal una masa no dolorosa, de consistencia dura, de superficie irregular y desplazable caudalmente. Anormalidades bioquímicas séricas incluyeron proteínas totales plasmáticas aumentadas, globulina y creatinina. Así, una laparotomía exploratoria fue realizada, identificando una masa en el mesenterio unida a la porción de intestino grueso. En el reporte se señalaron los siguientes hallazgos macroscópicos: una masa firme de 18 cm de longitud y 13 cm de ancho, con una superficie irregular, gran vascularización. Las muestras de tejido fueron fijadas en formalina buferada al 10% para análisis histopatológico e Inmunohistoquímica. Las muestras fueron procesadas por los métodos de rutina para microscopía y coloración con hematoxilina-eosina (HE), tricrómico de Masson, anticuerpos anti-citoqueratina CK5, anti-actina de músculo liso, vimentina y KIT(CD117). En conclusión, el tumor fue clasificado por histopatología como de origen mesenquimal e inmunopositividad citoplasmática difusa interna de células neoplásicas para actina de músculo liso, vimentina y negatividad para KIT, condujo al diagnóstico de leiomiosarcoma.

A Histological Study of Vasoconstriction by Local Anesthetics in Mandible

To assess the effect of epinephrine-containing local anesthetics on vasoconstriction, we immunohistochemically measured the intravascular lumen area in different regions of the mandible. Twelve male Wistar rats were used. General anesthesia was induced and maintained with sevoflurane. Infiltration anesthesia was performed with 0.2 mL of epinephrine-free 2% lidocaine (E-) near the left mandibular first molar and with 0.2 mL of epinephrine-containing 2% lidocaine (E+) near the right mandibular first molar. After decalcification, the specimens were paraffinized, and thin sections were prepared and immunohistologically stained with an antismooth muscle actin antibody. The intravascular lumen area was measured in the mucosa, periodontal membrane, Haversian/Volkmann's canal, and bone marrow. A Mann-Whitney U test was used for statistical processing, and p < .05 was considered to indicate a statistically significant difference. In the oral mucosa and the periodontal membrane, E+ had a significantly smaller vascular lumen area than E-. In the Haversian/Volkmann's canal and the bone marrow, no significant intergroup difference was observed in the intravascular lumen area. We postulate that this is due to a low smooth muscle content of blood vessels in the mandible and suggest that the vasoconstrictive effect of epinephrine-containing local anesthetics within the mandible is ineffective.

Retracted: Seropositivity and Titers of Anti-Smooth Muscle Actin Antibody Are Associated with Relapse of Type 1 Autoimmune Hepatitis

Retracted: Seropositivity and Titers of Anti-Smooth Muscle Actin Antibody Are Associated with Relapse of Type 1 Autoimmune Hepatitis

Seropositivity and Titers of Anti-Smooth Muscle Actin Antibody Are Associated with Relapse of Type 1 Autoimmune Hepatitis.

BackgroundIt is important to avoid relapse in autoimmune hepatitis (AIH) because repeated multiple relapses have been associated with a worse prognosis. However, risk factors for relapse before initiation of treatment are not fully understood. The aim of this study was to find predictive markers for relapse of type 1 AIH.Material/MethodsWe reviewed the records of 53 patients diagnosed with type 1 AIH based on the revised scoring system proposed by the International Autoimmune Hepatitis Group (IAIHG) between 2009 and 2014 at 4 hospitals belonging to the Saga Study Group of Liver Diseases (SASLD). We analyzed the differences in background characteristics between patients with or without relapse.ResultsAll patients achieved remission after treatment, and 9 (17%) subsequently relapsed. The relapsed patients were significantly younger and had a higher positive rate of anti-smooth muscle antibody (ASMA) than the non-relapsed patients (100% vs. 25%, P=0.0012). Moreover, relapse rate increased with titer of ASMA, while titer of antinuclear antibody was not associated with relapse rate.ConclusionsASMA is a useful predictive marker for relapse of type 1 AIH during or after withdrawal of medical therapy. More careful attention should be paid to immunosuppressive therapy in patients with high titers of ASMA.

Novel Association of miR-451 with the Incidence of TEVG Stenosis in a Murine Model.

The development of a tissue-engineered vascular graft (TEVG) holds great promise for advancing the field of cardiac surgery. Despite the successful translation of this technology, previous reports identify the primary mode of graft failure as stenosis secondary to intimal hyperplasia. MicroRNAs (miRNAs) regulate gene expression by interfering with mRNA function and recent research has suggested miRNA as a potential therapeutic target. The role of miRNAs in TEVGs during neotissue formation is currently unknown. In this study, we investigated if miRNAs regulate the inhibition of graft stenosis. Biodegradable PGA-P(LA/CL) scaffolds were implanted as inferior vena cava interposition grafts in a murine model (n = 14). Mice were sacrificed 14 days following implantation and TEVGs were harvested for histological analysis and miRNA profiling using Affymetrix miRNA arrays. Graft diameters were measured histologically, and the largest grafts (patent group) and smallest grafts (stenosed group) were profiled (n = 4 for each group). Cell population in each graft was analyzed with immunohistochemistry using antismooth muscle actin (SMA) and antimacrophage (F4/80) antibodies. The graft diameter was significantly greater in the patent group (0.63 ± 0.06 mm) than in the stenosed group (0.17 ± 0.06 mm) (p < 0.01). Cell proliferation was significantly greater in the stenosed grafts than in patent grafts (p < 0.01: SMA [187 ± 11 vs. 77 ± 8 cells] vs. p = 0.025: F4/80 [245 ± 23 vs. 187 ± 11 cells]). MiRNA array of 1416 genes showed that in stenosed grafts, mir-451, mir-338, and mir-466 were downregulated and mir-154 was upregulated. Mir-451 exhibited the greatest difference in expression between stenosed and patent grafts by -3.1-fold. Significant negative correlation was found between the expression of mir-451 and cell proliferation (SMA: r = -0.86, p = 0.003; F4/80: r = -0.89, p = 0.001). Our data, along with previous evidence that mir-451 regulates tumor suppressor genes, suggest that downregulation of mir-451 promotes acute proliferation of macrophages and smooth muscle cells, thereby inducing TEVG stenosis. Adequate expression of mir-451 may be critical for improving TEVG patency.

Therapeutic Treatment with Sustained-Release Platelet-Rich Plasma Restores Blood Perfusion by Augmenting Ischemia-Induced Angiogenesis and Arteriogenesis in Diabetic Mice

Objective: The objective of this investigation was to establish the effectiveness of sustained-release platelet-rich plasma (PRP) on perfusion and neovascularization in diabetic murine hind limb ischemia. Methods: After surgery in streptozotocin-induced diabetic mice, the mice were randomly assigned to the following 4 experimental groups: control (C), 100 µl of the sustained-release form of platelet-poor plasma (PPP), 100 µl of the solution form of PRP (PRP-sol), and 100 µl of the sustained-release form of PRP (PRP-sr). Endpoint evaluations were: blood perfusion by laser Doppler perfusion imaging (LDPI), vascular density by anti-vWF, and mature vessel density by anti-smooth muscle actin antibody. Results: This study demonstrated that a sustained release of PRP increases the perfusion of ischemic tissue as measured by LDPI (57 ± 12; 56 ± 9; 72 ± 7, and 98 ± 4 for the C, PPP, PRP-sol, and PRP-sr groups, respectively; p < 0.05), capillary density (151 ± 16; 158 ± 12; 189 ± 39, and 276 ± 39 for groups C, PPP, PRP-sol, and PRP-sr, respectively; p < 0.05), and mature vessel density (28 ± 2; 31 ± 3; 52 ± 10, and 85 ± 13 for the C, PPP, PRP-sol, and PRP-sr groups, respectively; p < 0.05). Conclusion: A sustained release of PRP containing potent angiogenic growth factors restores blood perfusion by stimulating angiogenesis and arteriogenesis.

The Study of Anatomic Relationship Between the Müller Muscle and the Tarsus in Asian Upper Eyelid

This study was performed to clarify the detailed anatomic relationship between the insertion of Müller muscle and the tarsus in Asian upper eyelids. Eight upper eyelids of 4 formalin-fixed Asian cadavers (4 males; age range, 48-69 years; mean age, 60.2 years) were examined. Three perpendicular dissected sections with a 2-mm thickness were obtained from each eyelid. One section was obtained from the midline, and another 2 sections were obtained, each 3-mm from the lateral and medial ends of the tarsus, respectively. Samples were stained with hematoxylin-eosin, Masson trichrome, and antismooth muscle actin antibody and examined microscopically. Müller muscle did not insert directly in the tarsus. However, its muscle tissues were tapered to the superior margin of the tarsus, and the space between the Müller muscle and the tarsus was bridged by fibrous connective tissue strands. Meanwhile, according to a result that measured distance between the Müller muscle insertion and the tarsus, the insertion of Müller muscle was positioned closer in the tarsus at the midline section than at the lateral or medial sections. This study suggests that the Müller muscle is broadly distributed around the superior border of the tarsus as tapered, aggregated smooth muscle bundles without direct insertion on the superior border of the tarsus, despite a few limitation.

Angiogenic properties of sustained release platelet-rich plasma: Characterization in-vitro and in the ischemic hind limb of the mouse

While single growth factor has limitation to induce optimal neovascularization, platelet-rich plasma (PRP) is an autologous reserver of various growth factors. However, little is known about the mechanism of PRP-related neovascularization.The objective of this investigation was to characterize the angiogenic and growth factor content of PRP and to determine, in vitro, its effect on endothelial cell proliferation. Additionally, this experiment sought to determine the effectiveness of different compositions of PRP (solution versus sustained release) on perfusion and neovascularization in a murine model of hind limb ischemia. Different growth factors were measured by enzyme-linked immunosorbent assay (ELISA). In vivo study, we used gelatin hydrogel as a sustained release carrier for growth factors in PRP. We induced hind limb ischemia by excising right femoral artery in wild type C57BL6 mice. After surgery, mice were randomly assigned to four experimental groups; control (C), 100 muL of sustained release form of platelet-poor plasma (PPP), 100 muL of solution form of PRP (PRP-sol), 100 muL of sustained release form of PRP (PRP-sr); each formulation was administered via an intramuscular injection to the ischemic hind limb. Endpoint evaluations were blood perfusion by laser Doppler perfusion image, vascular density by anti Von Willebrand factor (vWF), and mature vessel density by anti smooth muscle actin (SMA) antibody. Green fluorescent protein (GFP+) transgenic mice were generated by transplantation of bone marrow derived mononuclear cells to wild type C57BL6 mice, and finally CD34+ cell in the ischemic site of transgenic mice was detected by staining with anti-CD34 antibody. In vitro study showed that PRP containing different growth factors induces endothelial cell proliferation and capillary tube formation. In vivo study demonstrated that sustained release of PRP increased perfusion of ischemic tissue as measured by laser Doppler perfusion imaging (LDPI) (57 +/- 12, 56 +/- 9, 72 +/- 7, 98 +/- 4 for groups C, PPP, PRP-sol, and PRP-sr, respectively; P < .05); capillary density (151 +/- 16, 158 +/- 12, 189 +/- 39, 276 +/- 39 for groups C, PPP, PRP-sol, and PRP-sr, respectively, P < .05) and mature vessel density (28 +/- 2, 31 +/- 3, 52 +/- 10, 85 +/- 13 for groups C, PPP, PRP-sol, and PRP-sr, respectively, P < .05) . Sustained release PRP also increases CD34+ cells in the ischemic site of transgenic mice (6 +/- 3 vs 18 +/- 5/mm(2) for groups control and PRP-sr respectively, P < .05). Sustained release of PRP containing potent angiogenic growth factors restores blood perfusion presumably by stimulating angiogenesis, arteriogenesis, as well as vasculogenesis in the mouse hind limb ischemia. PRP is a natural reserver of various growth factors that can be collected autologously and is costeffective. Thus for clinical use, no special considerations concerning antibody formation and infection risk are needed.Some clinical devices to automatically prepare PRP are available at present. PRP are consistently being used clinically inthe department of orthopedics and plastic surgery (oral, maxillary facial) for a long time. On the basis of researchevidence, some publications have reported positive results in either bone or soft tissue healing. However, some researchconcludes that there is no or little benefit from PRP. This is likely due to faster degradation of growth factors in PRP sincesome authors suggest using sustained release form of PRP to deliver optimal effect of PRP. Gelatin hydrogel is also beingused clinically as a slow, sustained release of carrier for growth factors in our center recently.

Carcinoma in a Mixed Mammary Tumor in a Llama (Lama Glama)

A 13-year-old female llama was presented to the referring veterinarian for swelling and firmness of the right rear mammary gland, for a duration of 2 months, which had been unresponsive to antibiotics. A formalin-fixed wedge biopsy specimen from the affected quarter was submitted to Purdue University Animal Disease Diagnostic Laboratory for histopathology. Histopathologic examination revealed tubulopapillary acinar or solid nest-like clusters of neoplastic epithelial cells surrounded by whorls and sheets of proliferative myoepithelial cells. Histologic criteria for malignancy observed in neoplastic epithelial cells included marked cellular and nuclear atypia, high mitotic index, and numerous bizarre mitoses. The presence of osseous metaplasia in the proliferative mesenchymal component justified classification as a mixed tumor. Positive immunohistochemical staining of neoplastic epithelial cells with anticytokeratin antibody, and proliferative spindloid cells with antiviemtin and antismooth muscle actin antibodies supported the histopathologic diagnosis. The llama was in good health after about 1 year of initial presentation, and metastasis to regional lymph nodes was not reported. Mammary neoplasia is rare in camelids. To the authors' knowledge, this is the first report of a carcinoma in a mixed mammary tumor in a llama.

Léiomyome pelvien extra-utérin: diagnostic et prise en charge

To analyze the possibilities of setting up a therapy for extra-uterine pelvic leiomyomas.Three cases of leiomyomas of the broad ligament, of the round ligament and of the ovary, and literature review.Little is known about physiopathology of extra-uterine leiomyoma. The diagnosis of extra-uterine leiomyoma is based on histopathological analysis, using standard histology, and immunohistochemistry with anti-desmin and anti smooth muscle actin antibodies. The main differential diagnoses are fibroma, fibrothecoma, ovarian fibrosarcoma, and gastrointestinal stromal tumors. To define criteria of malignancy, we use Bell's classification without being sure that the uterine and extra-uterine models are comparable. So there is a risk of ignoring a low grade leiomyosarcoma. Providing therapy depends on the clinicopathologic features: the so called "parasitic leiomyoma", a tumor developed at the expense of local smooth muscle cells, metastasis of a benign metastasizing leiomyoma or leiomyomatosis peritonealis disseminata.The extra-uterine leiomyoma has no precise nosologic status and no specific criteria of benignity; thus no precise evolution can be predicted. We must be extremely careful, and the issue of the monitoring and long-term therapy of patients must come up.

Inhibition of Tumor Endothelial ERK Activation, Angiogenesis, and Tumor Growth by Sorafenib (BAY43-9006)

Activation of the Raf-MEK-ERK signal transduction pathway in endothelial cells is required for angiogenesis. Raf is the kinase most efficiently inhibited by the multikinase inhibitor sorafenib, which has shown activity against certain human cancers in clinical trials. To understand the mechanisms underlying this activity, we studied how it controlled growth of K1735 murine melanomas. Therapy caused massive regional tumor cell death accompanied by severe tumor hypoxia, decreased microvessel density, increased percentage of pericyte-covered vessels, and increased caliber and decreased arborization of vessels. These signs of K1735 angiogenesis inhibition, along with its ability to inhibit Matrigel neovascularization, showed that sorafenib is an effective anti-angiogenic agent. Extracellular signal-regulated kinase (ERK) activation in tumor endothelial cells, revealed by immunostaining for phospho-ERK and CD34, was inhibited, whereas AKT activation, revealed by phospho-AKT immunostaining, was not inhibited in K1735 and two other tumor types treated with sorafenib. Treatment decreased endothelial but not tumor cell proliferation and increased both endothelial cell and tumor cell apoptosis. These data indicate that sorafenib's anti-tumor efficacy may be primarily attributable to angiogenesis inhibition resulting from its inhibition of Raf-MEK-ERK signaling in endothelial cells. Assessing endothelial cell ERK activation in tumor bio-psies may provide mechanistic insights into and allow monitoring of sorafenib's activity in patients in clinical trials.

The Levator Aponeurosis Consists of Two Layers That Include Smooth Muscle

To investigate the two-fold structure of the levator aponeurosis, which is partly composed of independent smooth muscles. Fifteen upper eyelids of 12 Asian postmortems, with age at death ranging from 72 to 91 years, were examined. In 9 eyelids, posterior lamella tissue of the upper eyelid was removed to observe the stratified structures of the levator aponeurosis. Six full-thickness eyelids were used to observe the attachment site or the continuity between the levator aponeurosis and its surrounding tissues. The eyelids were incised perpendicularly in the center of the eyelid; samples were stained with Masson trichrome and antismooth muscle actin antibody and examined microscopically. Masson trichrome staining demonstrated the two-layered nature of the levator aponeurosis. The anterior layer was characterized by thick, robust fibrous tissue, and the posterior by thinner fibrous tissue. Although both layers contained muscle structures, the posterior layer contained more than the anterior. Immunostaining with antismooth muscle actin antibody revealed that the muscle in both layers was smooth muscle. The anterior layer continued to the orbital septum and the submuscular fibroadipose tissue; the posterior layers, located in front of Müller muscle and its tendon, attached to the anterior inferior one-third of the tarsus. Part of the anterior layer went through the orbicularis oculi muscle and attached to the subcuticular tissue. The levator aponeurosis is stratified, consisting of two layers than contain smooth muscle components in their proximal portions. It pulls mainly the preaponeurotic fat and anterior eyelid lamella. This partially regulates the tension of the eyelid and contributes to the ordered movement of the upper eyelid.

Forkhead Transcription Factors Inhibit Vascular Smooth Muscle Cell Proliferation and Neointimal Hyperplasia

Vascular smooth muscle cell (VSMC) proliferation and migration contribute significantly to atherosclerosis, postangioplasty restenosis, and transplant vasculopathy. Forkhead transcription factors belonging to the FoxO subfamily have been shown to inhibit growth and cell cycle progression in a variety of cell types. We hypothesized that forkhead proteins may play a role in VSMC biology. Under in vitro conditions, platelet-derived growth factor (PDGF)-BB, tumor necrosis factor-alpha, and insulin-like growth factor 1 stimulated phosphorylation of FoxO in human coronary artery smooth muscle cells via MEK1/2 and/or phosphatidylinositol 3-kinase-dependent signaling pathways. PDGF-BB, tumor necrosis factor-alpha, and insulin-like growth factor 1 treatment resulted in the nuclear exclusion of FoxO, whereas PDGF-BB alone down-regulated the FoxO target gene, p27(kip1), and enhanced cell survival and progression through the cell cycle. These effects were abrogated by overexpression of a constitutively active, phosphorylation-resistant mutant of the FoxO family member, TM-FKHRL1. The anti-proliferative effect of TM-FKHRL1 was partially reversed by small interfering RNA against p27(kip1). In a rat balloon carotid arterial injury model, adenovirus-mediated gene transfer of FKHRL1 caused an increase in the expression of p27(kip1) in the VSMC and inhibition of neointimal hyperplasia. These data suggest that FoxO activity inhibits VSMC proliferation and activation and that this signaling axis may represent a therapeutic target in vasculopathic disease states.

Oxytocin receptor expression in smooth muscle cells of peritoneal endometriotic lesions and ovarian endometriotic cysts

To investigate the expression of oxytocin receptor (OTR) in peritoneal and ovarian endometriotic lesions. Retrospective nonrandomized study. University hospital endometriosis research center. Premenopausal women with histologically confirmed endometriosis were selected. Peritoneal endometriotic lesions (n = 120); ovarian endometriotic cysts (n = 40); peritoneal biopsies, distant from the endometriotic lesion (n = 55); and unaffected peritoneal biopsies from patients without endometriosis (n = 11) were obtained. Hysterectomy specimens from patients without endometriosis and/or adenomyosis were used for controls (n = 10). Histopathological examination of peritoneal and ovarian specimens for OTR expression and identification of smooth muscle cells by immunohistochemistry staining with antibodies against OTR and smooth muscle actin. In addition, Western blot analysis, double-immunofluorescence, and in vitro studies with primary cell cultures have been performed. Comparison of the immunoreactive score of the OTR and smooth muscle actin expression with the smooth muscle content in peritoneum with and without endometriosis. In the epithelial cells of endometriotic lesions, we could demonstrate a high OTR expression. The stromal cells were OTR negative with the exception of some single cells. By using a monoclonal anti-smooth muscle actin antibody, these cells could be identified as intrastromal OTR-positive smooth muscle cells. The peritoneum of women with endometriosis shows a significantly higher smooth muscle content than the peritoneum of women without endometriosis. There were no significant differences between the smooth muscle content of active or inactive lesions and the stage of disease. Oxytocin receptor is expressed in smooth muscle cells and epithelial cells of peritoneal endometriotic lesions and ovarian endometriotic cysts. The inhibition of OTR by specific inhibitors might be a useful approach for the treatment of endometriosis-associated pain.