High Degree Of Vascularization Research Articles

BONE TISSUE REGENERATION INDUCED BY LOCAL DELIVERY OF BONE MORPHOGENETIC PROTEIN 2 FROM PRO-ANGIOGENIC NEAR-INFRARED-RESPONSIVE HYDROGELS

In this work, we combined tissue engineering and gene therapy technologies to develop a therapeutic platform for bone regeneration. We have developed photothermal fibrin-based hydrogels that incorporate degradable CuS nanoparticles (CuSNP) which transduce incident near-infrared (NIR) light into heat. A heat-activated and rapamycin-dependent transgene expression system was incorporated into mesenchymal stem cells to conditionally control the production of bone morphogenetic protein 2 (BMP-2). Genetically engineered cells were entrapped in the photothermal hydrogels. In the presence of rapamycin, photoinduced mild hyperthermia induced the release of BMP-2 from the NIR responsive cell constructs. Transcriptome analysis of BMP-2 expressing cells showed a signature of induced genes related to stem cell proliferation and angiogenesis. We next generated 4 mm diameter calvarial defects in the left parietal bone of immunocompetent mice. The defects were filled with NIR-responsive hydrogels entrapping cells that expressed BMP-2 under the control of the gene circuit. After one and eight days, rapamycin was administered intraperitoneally followed by irradiation with an NIR laser. Ten weeks after implantation, the animals were euthanized and samples from the bone defect zone were processed for histological analysis using Masson's trichrome staining and for immunohistochemistry analyses using specific CD31 and CD105 antibodies. Samples from mice that were only administered rapamycin or vehicle or that were only NIR-irradiated showed the persistence of fibrous tissue bridging the defect. In animals that were treated with rapamycin, NIR irradiation of implants resulted in the formation of new mineralized tissue with a high degree of vascularization, thus indicating the therapeutic potential of the approach.Acknowledgements: This research was supported by grants RTI2018-095159-B-I00 and PID2021-126325OB-I00 (MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”), by grant P2022/BMD- 7406 (Regional Government of Madrid). M.A.L-J. is the recipient of predoctoral fellowship PRE2019-090430 (MCIN/AEI/10.13039/501100011033).

CdTe quantum dots as fluorescent nanotools for in vivo glioblastoma imaging

Glioblastoma (GBM) is the most aggressive and infiltrating primary tumor of the central nervous system (CNS), showing a variety of mutations and a high degree of vascularity, cell polymorphism, and nuclear atypia. GBM treatment often recurs to surgical resection, but such protocol lacks efficacy since complete tumor removal is not entirely successful due to invasive cells that cannot be detected at the moment of the surgery. Here, we describe a new in vivo targeting and imaging method for GBM detection in an orthotropic mouse model using fluorescent CdTe quantum dots (CdTe QDs) conjugated to anti-glial fibrillary acidic protein (anti-GFAP). We conjugated and optimized red-emitting CdTe QDs to anti-GFAP to label GBM (U87 cell line) in vivo. The in vivo tumor growth was visualized by the hematoxylin and eosin staining and showed the successful delivery of GBM cells into the mouse brain parenchyma. CdTe/anti-GFAP QDs were injected into the tumor region, and their uptake by tumor cells was visualized by fluorescence microscopy, showing a specific dual labeling with vimentin-immunoreactive GBM. The results reported here provide new perspectives for using CdTe QDs in GBM detection, suggesting their potential application in imaging-guided surgery and a potential fluorescent tool to be applied in the monitoring of 3D tumor glial cultures.

RADT-35. STEREOTACTIC RADIOSURGERY FOR RESIDUAL, RECURRENT, AND METASTATIC HEMANGIOPERICYTOMAS: A SINGLE INSTITUTION EXPERIENCE

Abstract INTRODUCTION Hemangiopericytomas are infrequent vascular tumors originating from Zimmermann pericytes. The conventional treatment approach involves gross total resection (GTR), followed by adjuvant radiotherapy. Nevertheless, due to their tendency to infiltrate dural sinuses, high degree of vascularity, and anatomic inaccessibility, radical resection may be challenging with substantial risk of recurrence. To address this challenge, stereotactic radiosurgery (SRS) has been proposed as a promising adjuvant therapy. Objectives Our study aims to evaluate the effectiveness and safety of SRS as a potential treatment modality in patients with residual, recurrent, and metastatic hemangiopericytomas. METHODS From 1998 to 2023, 27 patients with 101 tumors underwent CyberKnife SRS at Stanford University Medical Center. The median age was 51 years at the time of treatment. The median follow-up period from SRS was 103 months (range: 6-250). All patients underwent upfront surgical resection. The median tumor volume was 1.5 cc. The median single-fraction equivalent dose (SFED) was 19 Gy. The SRS was administered at the 76% of the median isodose line (range: 64-89). RESULTS Of the 101 treated tumors, 24 (23.8%) progressed with a median time to recurrence of 30 months. At 10 years, the rates of local tumor control (LTC), overall survival (OS), and progression-free survival (PFS) were 74.3%, 80.8%, and 67%, respectively. In patients with residual lesions, the LTC rates were significantly greater when compared to those with residual or metastatic tumors. There was no significant difference between patients with residual, recurrent, and metastatic hemangiopericytomas in OS and PFS. Notably, no cases of radiation-induced adverse events were detected. CONCLUSION Our study provides the largest single-institutional retrospective series with hemangiopericytomas treated with SRS. SRS leads to excellent LTC, PFS, and OS at 10 years with negligible risk for adverse events. Therefore, it is an effective and safe management modality for patients with residual, recurrent, and metastatic hemangiopericytomas.

ROLE OF PREOPERATIVE EMBOLIZATION IN BONE TUMORS- A case control study

Preoperative embolization of bone tumor is a minimally invasive angiographic intervention technique and effective adjuvant treatment option for bone tumors. In 1975, Feldman was the rst to use the pre-operative arterial embolization technique to downstage the vascularity of bone tumors, thus reducing blood loss during surgery. Surgery remains the mainstay treatment option for bone tumors. Embolization reduces tumor vascularity and hence decreases intraoperative blood loss and intraoperative blood transfusion. It enables more accurate delineation of tissue planes during surgery, resulting in complete removal of the bone tumor. The most frequent bone tumors that typically need preoperative embolization are those difcult to operate on because of their huge size, high degree of vascularity, proximity to critical tissues, or difcult surgical access. PVA particles are regarded as a workhorse in preoperative transarterial embolization and thus used commonly as an embolizing agent. It is safe procedure and therefore can be used as part of the multidisciplinary approach to the management highly vascular bone tumors.

Core-Shell Structured Porous Calcium Phosphate Bioceramic Spheres for Enhanced Bone Regeneration.

Adequate new bone regeneration in bone defects has always been a challenge as it requires excellent and efficient osteogenesis. Calcium phosphate (CaP) bioceramics, including hydroxyapatite (HA) and biphasic calcium phosphates (BCPs), have been extensively used in clinical bone defect filling due to their good osteoinductivity and biodegradability. Here, for the first time, we designed and fabricated two porous CaP bioceramic granules with core-shell structures, named in accordance with their composition as BCP@HA and HA@BCP (core@shell). The spherical shape and the porous structure of these granules were achieved by the calcium alginate gel molding technology combined with a H2O2 foaming process. These granules could be stacked to build a porous structure with a porosity of 65-70% and a micropore size distribution between 150 and 450 μm, which is reported to be good for new bone ingrowth. In vitro experiments confirmed that HA@BCP bioceramic granules could promote the proliferation and osteogenic ability when cocultured with bone marrow mesenchymal stem cells, while inhibiting the differentiation of RAW264.7 cells into osteoclasts. In vivo, 12 weeks of implantation in a critical-sized femoral bone defect animal model showed a higher bone volume fraction and bone mineral density in the HA@BCP group than in the BCP@HA or pure HA or BCP groups. From histological analysis, we discovered that the new bone tissue in the HA@BCP group was invading from the surface to the inside of the granules, and most of the bioceramic phase was replaced by the new bone. A higher degree of vascularization at the defect region repaired by HA@BCP was revealed by 3D microvascular perfusion angiography in terms of a higher vessel volume fraction. The current study demonstrated that the core-shell structured HA@BCP bioceramic granules could be a promising candidate for bone defect repair.

Isolinderalactone inhibits glioblastoma cell supernatant-induced angiogenesis.

Glioblastoma multiforme (GBM) is the most frequently occurring malignant brain tumor in adults and is characterized by a high degree of vascularization. Glioblastoma cells communicate with their microenvironment and stimulate blood vessel formation to support tumor progression. It has previously been reported that isolinderalactone induces apoptosis in GBM cells and suppresses the growth of glioblastoma xenograft tumors in vivo. Furthermore, isolinderalactone has been shown to inhibit the hypoxia-driven upregulation of vascular endothelial growth factor (VEGF) in U-87 GBM cells and strongly reduce VEGF-triggered angiogenesis in vitro and in vivo. In the present study, the direct angiogenic effect of GBM and the effect of isolinderalactone on tumor angiogenesis were investigated. Culture supernatants were obtained from U-87 cells under normoxic or hypoxic conditions to provide normoxic conditioned medium (NCM) and hypoxic conditioned medium (HCM) respectively. The NCM and HCM were each used to treat to human brain microvascular endothelial cells (HBMECs), and their effects were observed using wounding migration and tube formation assays. HCM increased the migration and capillary-like tube formation of HBMECs when compared with NCM, and treatment with isolinderalactone suppressed the HCM-driven angiogenesis in vitro. Additionally, isolinderalactone decreased HCM-triggered angiogenic sprouting in HBMECs in a 3D microfluidic device after the application of an HCM-containing interstitial fluid flow. Furthermore, isolinderalactone strongly reduced HCM-triggered angiogenesis in an in vivo Matrigel plug assay in mice. These findings provide evidence of angiogenesis inhibition by isolinderalactone, and demonstrate the anti-angiogenic effect of isolinderalactone against the direct angiogenic effect of GBM tumor cells.

Ascorbic acid 2-glucoside preconditioning enhances the ability of bone marrow mesenchymal stem cells in promoting wound healing

BackgroundNowadays, wound is associated with a complicated repairing process and still represents a significant biomedical burden worldwide. Bone marrow mesenchymal stem cells (BMSCs) possess multidirectional differentiation potential and secretory function, emerging as potential cellular candidates in treating wounds. Ascorbic acid 2-glucoside (AA2G) is a well-known antioxidant and its function in BMSC-promoting wound healing is worth exploring.MethodsThe in vitro cell proliferation, migration, and angiogenesis of BMSCs and AA2G-treated BMSCs were detected by flow cytometry, EDU staining, scratch assay, transwell assay, and immunofluorescence (IF). Besides, the collagen formation effect of AA2G-treated BMSCs conditioned medium (CM) on NIH-3T3 cells was evaluated by hydroxyproline, qRT-PCR and IF staining detection. Next, in the wound healing mouse model, the histological evaluation of wound tissue in PBS, BMSCs, and AA2G-treated BMSCs group were further investigated. Lastly, western blot and ELISA were used to detect the expression levels of 5-hmc, TET2 and VEGF protein, and PI3K/AKT pathway activation in BMSCs treated with or without AA2G.ResultsThe in vitro results indicated that AA2G-treated BMSCs exhibited stronger proliferation and improved the angiogenesis ability of vascular endothelial cells. In addition, the AA2G-treated BMSCs CM enhanced migration and collagen formation of NIH-3T3 cells. In vivo, the AA2G-treated BMSCs group had a faster wound healing rate and a higher degree of vascularization in the new wound, compared with the PBS and BMSCs group. Moreover, AA2G preconditioning might enhance the demethylation process of BMSCs by regulating TET2 and up-regulating VEGF expression by activating the PI3K/AKT pathway.ConclusionsAA2G-treated BMSCs promoted wound healing by promoting angiogenesis and collagen deposition, thereby providing a feasible strategy to reinforce the biofunctionability of BMSCs in treating wounds.

Usefulness of cell-penetrating peptides and penetration accelerating sequence for nose-to-brain delivery of glucagon-like peptide-2

Neuropeptides are expected as therapeutic drug candidates for central nervous system (CNS) disorders. Intracerebroventricular (i.c.v.) administration of glucagon-like peptide-2 (GLP-2) has an antidepressant-like effect not only in depression model mice but also in treatment-resistant depression model mice. However, because i.c.v. administration is very invasive, research is progressing on brain delivery using intranasal administration as a non-invasive method. After intranasal administration of the drug, there are two routes to the brain. That of direct delivery from the paracellular route of olfactory epithelium to the brain via the olfactory bulb has been studied, and that of systemic absorption via the paracellular route of respiratory epithelium has been put to practical use. The high degree of vascularization and permeability of the nasal mucosa enables drug delivery via the paracellular route that leads to systemic delivery. Therefore, suppressing systemic absorption may increase drug delivery to brain, so we focused on the transcellular route. We created a GLP-2 derivative by adding cell-penetrating peptides (CPP) and penetration accelerating sequences (PAS), which are reported to provide efficient intracellular uptake, to GLP-2. However, to deliver GLP-2 by the transcellular route, GLP-2 must not only be taken up into cells but also move out of the cells. We investigated in vitro and in vivo function of PAS-CPP-GLP-2 to enable the translocation of GLP-2 directly from the nose to the brain. Derivatization of PAS-CPP-GLP-2 prevented its degradation. In the evaluation of intracellular dynamics, PAS-CPP-GLP-2 enhanced cellular uptake by macropinocytosis with CPP and promoted escape from endosomal vesicles by PAS. This study also showed that PAS-CPP-GLP-2 can move out of cells. Furthermore, only this PAS-CPP-GLP-2 showed an antidepression-like effect within 20 min of intranasal administration. Intranasal administered PAS-CPP-GLP-2 surprisingly showed the effect at the same dose with i.c.v. administration, but intravenous administered PAS-CPP-GLP-2 did not show the effect. These results suggested that PAS-CPP-GLP-2 can be efficiently delivered from the nose to the CNS and show a pharmacological effect, demonstrating the usefulness of PAS and CPP for nose-to-brain delivery of GLP-2.

Evaluation of decellularization protocols for production of porcine small intestine submucosa for use in abdominal wall reconstruction.

Porcine-derived acellular biologic grafts are increasingly used in abdominal wall reconstruction and other soft tissue repairs. In a previous work, we have shown porcine small intestine submucosa (PSIS) exhibits clear advantages over porcine pericardium (PPC) and porcine acellular dermal matrix (PADM) in repairing full-thickness abdominal wall defects. In the present study, we aim to determine, quantify, and compare the effects of two most commonly used decellularization protocols on biomechanical and biocompatible properties of PSIS. After mechanical preparation, PSIS was treated with either alkaline and acid (AA) protocol or sodium dodecyl sulfate (SDS) protocol. Cellular content removal, preservation of matrix components, micro- and ultra- structures, and mechanical properties were compared. The host responses were evaluated using PSIS for repairing rat abdominal wall defects. With regard to the absence of cellular contents, neatly arranged collagen fiber structures, better retention of growth factors, better mechanical strength, lower degrees of local and systemic inflammatory responses, higher degree of vascularization and tissue ingrowth, alkaline and acid protocol exhibits clear advantages over SDS protocol for the preparation of PSIS extracellular matrix.

Injectable tissue integrating networks from recombinant polypeptides with tunable order.

Emergent properties of natural biomaterials result from the collective effects of nanoscale interactions among ordered and disordered domains. Here, using recombinant sequence design, we have created a set of partially ordered polypeptides to study emergent hierarchical structures by precisely encoding nanoscale order-disorder interactions. These materials, which combine the stimuli-responsiveness of disordered elastin-like polypeptides and the structural stability of polyalanine helices, are thermally responsive with tunable thermal hysteresis and the ability to reversibly form porous, viscoelastic networks above threshold temperatures. Through coarse-grain simulations, we show that hysteresis arises from physical crosslinking due to mesoscale phase separation of ordered and disordered domains. On injection of partially ordered polypeptides designed to transition at body temperature, they form stable, porous scaffolds that rapidly integrate into surrounding tissue with minimal inflammation and a high degree of vascularization. Sequence-level modulation of structural order and disorder is an untapped principle for the design of functional protein-based biomaterials.

Selection of optimum modes of the Nd:YAG-laser with the wavelength of 1064 nm for the surgical treatment of the juvenile nasopharyngeal angiofibroma

Juvenile nasopharyngeal angiofibroma is a benign tumor with a high degree of vascularization. It has a local form of expansive growth. Abundance of a blood vessels and angiomatous structure of embryonic vessels lead to massive bleeding after the damage or removal of the tumor. For the prevention of an abundant bleeding the preoperative procedures are conducted as well as the different intraoperative methods. One of these methods is using the surgical laser. We took a morphological research of the removed tumor tissue to select the optimum parameters of the Nd:YAG laser with the wavelength of 1064 nm. According to the results of histological examination the effective laser mode has been determined for the surface and interstitial photocoagulation.

ФЕНО- И ГЕНОТИПИЧЕСКИЕ ХАРАКТЕРИСТИКИ НЕДИФФЕРЕНЦИРОВАННОЙ ДИСПЛАЗИИ СОЕДИНИТЕЛЬНОЙ ТКАНИ

The aim of the study was to study the clinical-anamnestic, molecular-genetic and morphological characteristics of patients with tubal-peritoneal infertility on the background of undifferentiated connective tissue dysplasia (uCTD). Material and methods . 60 cases of tubal-peritoneal infertility were analyzed. Patients were divided into 2 groups on the basis of a modified scoring scale by T. I. Kadurina. The degree of expression of uCTD on a modified scale by T. I. Kadurina and on the scale of T. I. Kadurina and L.N. Abbamukova was evaluated. In the following, the patients were genotyped for carriage of IL6-174 C / G gene polymorphism [rs1800795] and VEGF 634 G / C [rs2010963]. Some patients, from the groups represented, under operative intervention, performed a biopsy of fragments of the aponeurosis of the anterior abdominal wall followed by hemostasis. To study the structure of the connective tissue of the extracellular matrix, the sections were stained by Van-Geison and by Mallory. In the samples collagens I and III, as well as CD34 + -structures were detected immunohistochemically. Results. Using strict criteria (more than 6 external phenotypes), the clinical significance of anomalies increases, and the frequency decreases to 25%. Clinical manifestations of signs of hereditary pathology of connective tissue depend on environmental factors, gene penetrance and patterns of gene expression. Conclusions. Diagnosis of uCTD is possible already at the stage of physical examination with a complex evaluation of external and visceral phenotypic markers on the scale of T. I. Kadurina and L.N. Abbamukova and on a modified scale by T. I. Kadurina. The polymorphism of the genes IL6 - 174 [rs1800795] C> G C / C and VEGFA 634 [rs2010963] G> C G / C is a predictor of uCTD. Expression of type III collagen in this pathology indicates the presence of an unformed connective tissue, characterized by a high degree of vascularization. Violation of the architectonics of connective tissue is the basis for the formation of a multifaceted dysplasia associated pathology in patients with tubal-peritoneal infertility.

In Vitro Human Umbilical Vein Endothelial Cells Response to Ionic Dissolution Products from Lithium-Containing 45S5 Bioactive Glass.

Since lithium (Li+) plays roles in angiogenesis, the localized and controlled release of Li+ ions from bioactive glasses (BGs) represents a promising alternative therapy for the regeneration and repair of tissues with a high degree of vascularization. Here, microparticles from a base 45S5 BG composition containing (wt %) 45% SiO2, 24.5% Na2O, 24.5% CaO, and 6% P2O5, in which Na2O was partially substituted by 5% Li2O (45S5.5Li), were obtained. The results demonstrate that human umbilical vein endothelial cells (HUVECs) have greater migratory and proliferative response and ability to form tubules in vitro after stimulation with the ionic dissolution products (IDPs) of the 45S5.5Li BG. The results also show the activation of the canonical Wnt/β-catenin pathway and the increase in expression of proangiogenic cytokines insulin like growth factor 1 (IGF1) and transforming growth factor beta (TGFβ). We conclude that the IDPs of 45S5.5Li BG would act as useful inorganic agents to improve tissue repair and regeneration, ultimately stimulating HUVECs behavior in the absence of exogenous growth factors.

Cavernous hemangioma of the bladder in a child of 6 years and 11 months

We analyze a clinical case of the patient at the age of 6 years and 11 months with a cavernous hemangioma of the bladder. The disease manifested with isolated macrohaematuria. During the diagnostic assessment by 3D-ultrasound examination of the urinary tract with tight bladder filling, a mass with tuberous shape and echonegative areas inside was visualized on the left bladder wall close to its anterior surface, while external bladder contours were unchanged. Color Doppler mapping showed some vessels in the projection of the mass sprouting from the muscular layer of the bladder. The neoplasm was confirmed by abdominopelvic magnetic resonance imaging (with and without contrast enhancement) and by cystoscopy. Taking into account familial cancer history of the patient (oncological disorders in the maternal and paternal lines) and high degree of vascularization of the mass, a malignancy (rhabdomyosarcoma) was deemed highly probable. At 2 months after the disease manifestation, transurethral endoscopic surgery was performed with coagulation and a resection of an exophytic mass of 1 cm in diameter, with a wide base, irregular tuberous shape, brown colored, with a cystic component. The mucous layer on the lateral bladder wall was abnormal with some hemorrhages. Histological examination of the tissue sample showed a cavernous hemangioma, that was the final diagnosis in this patient. During subsequent 2-year follow up, there was no recurrence of the disease. The described case is of clinical interest because bladder hemangiomas are rare in children; therefore, this disease should be on the list for differential diagnosis in children with macrohaematuria.

Confocal raman microspectral imaging of ex vivo human spinal cord tissue

Confocal Raman microspectral imaging (CRMI) provides a versatile tool to illustrate the biochemical nature and structure of biological tissue without introducing any external labels. In this work, a precise correlation was established between the biochemical profile and histological architecture of ex vivo human spinal cord tissue by using CRMI with 633nm excitation. After precisely linking the spectral features to the chemical constituents, much information about the molecular composition of both gray and white matter were revealed. Two-dimensional Raman images were generated by integrating the intensities of the characteristic Raman bands in the area of the intermediate column and ventral horn. K-mean cluster analysis was further applied to visualize the underlying morphological basis of spinal cord tissue by chemical component types and their distribution pattern. Lipid-rich white matter could be visually distinguished from gray matter considering a CH2 bending/scissoring band at 1445cm−1 and an amide III band at 1250cm−1. Meanwhile, the formation and distribution pattern of perineuronal nets (PNNs) in the scanning area was validated by the integration of saccharides (617cm−1) and amide III bands. Moreover, the heme profile indicated a higher degree of vascularization in gray matter. All of the results obtained testified to the possibility that gray matter could be more susceptible to spinal cord injury (SCI) because of capillary network distribution and glycosaminoglycans (GAGs) aggregation. These findings are important for interpreting the morphological specificity of human spinal cord tissue, and also for studying the molecular basis of SCI.

Role of preoperative embolization for intradural spinal hemangioblastomas

Spinal hemangioblastomas (HB) are relatively rare neoplasms with a high degree of vascularity. Therapy for symptomatic tumors involves total resection when possible. Due to the enriched blood supply of these neoplasms, there is a high risk of significant intraoperative blood loss, which can lead to perioperative complications. Preoperative embolization of HB has been suggested to reduce blood loss and operative morbidity, but its use remains controversial. Data on the risks and benefits of preoperative embolization for this tumor remains limited. We identified and analyzed all 29 reported cases of preoperative embolization of intradural spinal HB within the literature. There were 18 men and nine women, and patients ranged from 24 to 61years of age. Mean tumor size was 3.5cm. Cervical and thoracic location was most common, accounting for 48.3% and 20% of cases, respectively. Complications from embolization and surgery were minimal, with no deaths or permanent neurological morbidity. Minimal intraoperative bleeding and excellent rates of gross total resection were reported with preoperative embolization. However, outcomes from microsurgery alone from historical series have similarly reported excellent outcomes. While there is no established standard, preoperative embolization should be reserved for particularly high risk patients with risk of intraoperative bleeding.

Pharmacokinetic Properties of a New Glutamic Acid Derivative Glutaron.

Inhomogenous distribution of glutaron in organs and tissues was found after intravenous and peroral administration: the agent demonstrated high affinity to organs with high degree of vascularization (lungs and heart) and elimination (kidney). Glutaron easily penetrates through the blood-brain barrier, which is consistent with its concentration in the adipose tissue.

Identification of the Lipodepsipeptide MDN-0066, a Novel Inhibitor of VHL/HIF Pathway Produced by a New Pseudomonas Species.

Throughout recent history, metabolites of microbial origin have had an extraordinary impact on the welfare of humanity. In fact, natural products have largely been –and still are– considered an exceedingly valuable platform for the discovery of new drugs against diverse pathologies. Such value is partly due to their higher complexity and chemical diversity as compared to those of synthetic and combinatorial compounds. Mutations in the Von Hippel-Lindau (vhl) gene are responsible for VHL disease, congenital polycythemia, and are found in many sporadic tumor types. The primary cause of morbidity and mortality for these patients arises from progression of Renal Cell Carcinoma (RCC) or end-stage renal disease. Inactivation of the Von Hippel-Lindau (vhl) tumor suppressor gene arises in the majority of Renal Cell Carcinoma (RCC) as well as in other types of cancer and is associated with a high degree of vascularization and poor prognosis. Loss of pVHL function thus represents a pathognomonic molecular defect for therapeutic exploitation. In this study, renal carcinoma cell lines with naturally occurring vhl mutations (RCC4 VA) and their genetically matched wild-type vhl (RCC4 VHL) counterparts were seeded onto 96-well plates and treated with a collection of 1,040 organic extracts obtained from 130 bacterial strains belonging to at least 25 genera of the phyla Actinobacteria, Firmicutes, Proteobacteria and Bacteroidetes. This strategy allowed us to identify several extracts obtained from bacterial strain F-278,770T, the type strain of the recently proposed new species Pseudomonas granadensis, showing biological activities not associated with previously known bioactive metabolites. The fractionation and structural elucidation of one of these extracts led to the discovery of a new lipodepsipeptide (MDN-0066) with specific toxicity in pVHL deficient cells that is not detectable in cells with pVHL expression rescue. This specific toxicity is associated with apoptosis induction in VHL deficient cell line as demonstrated with PARP activation and Annexin V staining. Our study demonstrated the feasibility of selectively targeting the loss of the vhl tumor suppressor gene for potential clinical benefit. Our results may have great impact on the development of new targeted therapies from natural products for the treatment of cancer and other genetic diseases.

Eccrine spiradenoma: a case report

Eccrine spiradenoma is a rare benign adnexal epithelial neoplasm characterized by a bluish-pink, slow-growing solitary nodule, coursing with paroxysmal pain, mainly in young adults. It is most commonly located in the trunk and extremities. Bleeding and/or ulceration are common in the giant vascular variant, which is characterized by a size greater than 2 cm in diameter and a high degree of vascularization seen on histology. Its clinical characteristics are unspecific and require biopsy with histological analysis for diagnosis. Its histology presents one or more basophilic intradermal lobules enveloped in a fibrous capsule without connection to the epidermis. It presents two types of cells: small cells with dark nuclei in the periphery and large cells with light nuclei in the center. An accurate diagnosis is important due to the fact that it can develop into a potentially fatal malignancy – which is a rare phenomenon and occurs most often in the multiple variant. A rare case of eccrine spiradenoma in the abdomen with bleeding associated is described in the present paper, as well as a discussion on possible treatments.

Novel approach of MALDI drug imaging, immunohistochemistry, and digital image analysis for drug distribution studies in tissues.

Drug efficacy strongly depends on the presence of the drug substance at the target site. As vascularization is an important factor for the distribution of drugs in tissues, we analyzed drug distribution as a function of blood vessel localization in tumor tissue. To explore distribution of the anticancer drugs afatinib, erlotinib, and sorafenib, a combined approach of matrix-assisted laser desorption/ionization (MALDI) drug imaging and immunohistochemical vessel staining was applied and examined by digital image analysis. The following two xenograft models were investigated: (1) mice carrying squamous cell carcinoma (FaDu) xenografts (ntumor = 13) were treated with afatinib or erlotinib, and (2) sarcoma (A673) xenograft bearing mice (ntumor = 8) received sorafenib treatment. MALDI drug imaging revealed a heterogeneous distribution of all anticancer drugs. The tumor regions containing high drug levels were associated with a higher degree of vascularization than the regions without drug signals (p < 0.05). When correlating the impact of blood vessel size to drug abundance in the sarcoma model, a higher amount of small vessels was detected in the tumor regions with high drug levels compared to the tumor regions with low drug levels (p < 0.05). With the analysis of coregistered MALDI imaging and CD31 immunohistochemical data by digital image analysis, we demonstrate for the first time the potential of correlating MALDI drug imaging and immunohistochemistry. Here we describe a specific and precise approach for correlating histological features and pharmacokinetic properties of drugs at microscopic level, which will provide information for the improvement of drug design, administration formula or treatment schemes.