Periphery Of Lesion Research Articles

Evidence of Resting-state Activity in Propofol-anesthetized Patients with Intracranial Tumors

Resting-state (RS) networks, revealed by functional magnetic resonance imaging (fMRI) studies in healthy volunteers, have never been evaluated in anesthetized patients with brain tumors. Our purpose was to examine the presence of residual brain activity on the auditory network during propofol-induced loss of consciousness in patients with brain tumors. Twenty subjects with intracranial masses were prospectively studied by means of intraoperative RS-fMRI acquisitions before any craniectomy. After performing single-subject independent component analysis, spatial maps and time courses were assigned to an auditory RS network template from the literature and compared via spatial regression coefficients. All fMRI data were of sufficient quality for further postprocessing. In all but two patients, the RS functional activity of the auditory network could be successfully mapped. In almost all patients, contralateral activation of the auditory network was present. No significant difference was found between the mean distance of the RS activity clusters and the lesion periphery for tumors located in the temporal gyri vs. those in other brain regions. The spatial deviation between the activated cluster in our experiment and the template was significantly (P = 0.04) higher in patients with tumors located in the temporal gyri than in patients with tumors located in other regions. Propofol-induced anesthesia in patients with intracranial lesions does not alter the blood-oxygenation level-depended signal, and independent component analysis of intraoperative RS-fMRI may allow assessment of the auditory network in a clinical setting.

En face enhanced depth imaging optical coherence tomography of polypoidal choroidal vasculopathy

PurposeTo analyse retinal and choroidal changes associated with polypoidal choroidal vasculopathy (PCV) using en face spectral-domain optical coherence tomography (SD-OCT).MethodsIn this retrospective and descriptive study, we collected imaging of patients...

MRI characteristics of early PML-IRIS after natalizumab treatment in patients with MS

ObjectiveThe early detection of MRI findings suggestive of immune reconstitution inflammatory syndrome (IRIS) in natalizumab-associated progressive multifocal leukoencephalopathy (PML) is of crucial clinical relevance in terms of treatment decision-making and...

Hepatocellular carcinoma: stiffness value and ratio to discriminate malignant from benign focal liver lesions.

To investigate the use of stiffness value and stiffness ratio (ratio of lesion to background liver parenchyma values) to discriminate malignant from benign focal liver lesions by using histologic results as the reference standard. This study was approved by the institutional review board, and written informed consent was obtained. Three hundred seventy-three patients with focal liver lesions proven at histologic examination underwent measurement of liver stiffness with elastography point quantification. First, stiffness values in two regions of the background liver parenchyma (at 0.5-2 cm and >2 cm from the lesion periphery) near 163 hepatocellular carcinomas were analyzed to determine a reference background liver for calculating the stiffness ratio. Second, the use of the lesion stiffness value and the stiffness ratio for prediction of liver malignancy was investigated in a cohort of patients with 58 benign and 201 malignant lesions. Results were validated in another independent cohort of patients with 25 benign and 89 malignant lesions by using analysis of the area under the receiver operating characteristic (AUC) curve. The coefficient of variation for the background liver at 0.5-2 cm from the lesion was higher (196%) than that at greater than 2 cm from the lesion (66%). In the development phase, diagnostic accuracy with use of the stiffness value was significantly higher than that with use of the stiffness ratio for discrimination of malignant from benign lesions (AUC, 0.86 vs 0.66, respectively; P < .001). Diagnostic performance with the stiffness value was lower than that with the stiffness ratio (AUC, 0.53 vs 0.86, respectively; P < .001) for discrimination of cirrhotic nodules from other benign lesions. Diagnostic performance with the stiffness value was significantly lower than that with the stiffness ratio (AUC, 0.58 vs 0.71 respectively; P = .007) for discrimination of metastasis from primary liver cancers. In the validation phase, similar findings were revealed for the discrimination of malignant from benign lesions (AUC, 0.87 vs 0.67; P < .001) and discrimination between metastasis and primary liver cancers (AUC, 0.49 vs 0.73; P < .001). Use of stiffness values measured in the liver parenchyma at more than 2 cm from the lesion allowed better diagnostic performance than did values measured in a region closer to the tumor. Stiffness value was more accurate than stiffness ratio for differentiation of malignant from benign focal liver lesions, but the stiffness ratio might be useful for subclassification of benign and malignant lesions. Online supplemental material is available for this article.

A correlation of computed tomography perfusion and histopathology in tumor edges of hepatocellular carcinoma

The peripheral morphologic characteristics of hepatocellular carcinoma (HCC) reflect tumor growth patterns. Computed tomography (CT) perfusion is a new method to analyze hemodynamic changes in tissues. We assessed the relationship between CT perfusion and histopathologic findings in the periphery of HCC lesions. Non-contrast CT, enhanced dual-phase CT, and CT perfusion were performed on 77 subjects (47 patients and 30 controls). Based on the imaging findings of enhanced dual-phase CT, the tumor edges were classified into three types: type I (sharp); type II (blurry); and type III (mixed). The CT perfusion parameters included hepatic blood flow, hepatic arterial fraction, hepatic arterial perfusion, and hepatic portal perfusion. The tissue sections from resected specimens were subjected to routine hematoxylin and eosin staining and immunohistochemical staining for CD34. The correlations between microvessel density (MVD) and the CT perfusion parameters were analyzed using Pearson's product-moment correlation coefficient. Changes in the perfusion parameters in tumor edges of different tumor types were evaluated. Type I (sharp): the pathologic findings showed fibrous connective tissue capsules in the tumor edges, and an MVD ≤30/mm2. Type II (blurry): the histology showed that the edges were clear with no capsules and an MVD>30/mm2. Type III (mixed): the pathology was similar to that of types I and II, and an MVD>30/mm2. Hepatic blood flow, hepatic arterial fraction, hepatic arterial perfusion, and hepatic portal perfusion were significantly increased in the tumor edges of HCC patients compared to those of the controls (P<0.05). The correlation between CT perfusion parameters and MVD was higher in blurry tumor edges of type II than in those of types I or III. CT perfusion imaging of tumor edges may be helpful in revealing histopathological features, and indirectly reflect angiogenic changes of HCCs.

Guanosine Protects Against Cortical Focal Ischemia. Involvement of Inflammatory Response.

Stroke is the major cause of death and the most frequent cause of disability in the adult population worldwide. Guanosine plays an important neuroprotective role in several cerebral ischemic models and is involved in the modulation of oxidative responses and glutamatergic parameters. Because the excessive reactive oxygen species produced during an ischemic event can trigger an inflammatory response, the purpose of this study was to evaluate the hypothesis that guanosine is neuroprotective against focal cerebral ischemia, inhibits microglia/macrophages activation, and mediates an inflammatory response ameliorating the neural damage. Permanent focal cerebral ischemia was induced in adult rats, and guanosine was administered immediately, 1, 3, and 6 h after surgery. Twenty-four hours after ischemia, the asymmetry scores were evaluated by the cylinder test; neuronal damage was evaluated by Fluoro-Jade C (FJC) staining and propidium iodide (PI) incorporation; microglia and immune cells were evaluated by anti-Iba-1 antibody; and inflammatory parameters such as interleukins (IL): IL-1, IL-6, IL-10; tumor necrosis factors alpha (TNF-α); and interferon-gamma (INF-γ) were evaluated in the brain tissue and cerebrospinal fluid. The ischemic event increased the levels of Iba-1-positive cells and pro-inflammatory cytokines and decreased IL-10 levels (an anti-inflammatory cytokine) in the lesion periphery. The guanosine treatment attenuated the changes in these inflammatory parameters and also reduced the infarct volume, PI incorporation, and number of FJC-positive cells, improving the functional recovery. Thus, guanosine may have been a promising therapeutic agent for the treatment of ischemic brain injury by reduction of inflammatory process triggered in an ischemic event.

Runx1 Activities in Superficial Zone Chondrocytes, Osteoarthritic Chondrocyte Clones and Response to Mechanical Loading.

Runx1, the hematopoietic lineage determining transcription factor, is present in perichondrium and chondrocytes. Here we addressed Runx1 functions, by examining expression in cartilage during mouse and human osteoarthritis (OA) progression and in response to mechanical loading. Spared and diseased compartments in knees of OA patients and in mice with surgical destabilization of the medial meniscus were examined for changes in expression of Runx1 mRNA (Q-PCR) and protein (immunoblot, immunohistochemistry). Runx1 levels were quantified in response to static mechanical compression of bovine articular cartilage. Runx1 function was assessed by cell proliferation (Ki67, PCNA) and cell type phenotypic markers. Runx1 is enriched in superficial zone (SZ) chondrocytes of normal bovine, mouse, and human tissues. Increasing loading conditions in bovine cartilage revealed a positive correlation with a significant elevation of Runx1. Runx1 becomes highly expressed at the periphery of mouse OA lesions and in human OA chondrocyte 'clones' where Runx1 co-localizes with Vcam1, the mesenchymal stem cell (MSC) marker and lubricin (Prg4), a cartilage chondroprotective protein. These OA induced cells represent a proliferative cell population, Runx1 depletion in MPCs decreases cell growth, supporting Runx1 contribution to cell expansion. The highest Runx1 levels in SZC of normal cartilage suggest a function that supports the unique phenotype of articular chondrocytes, reflected by upregulation under conditions of compression. We propose Runx1 co-expression with Vcam1 and lubricin in murine cell clusters and human 'clones' of OA cartilage, participate in a cooperative mechanism for a compensatory anabolic function.

Significance of CD123 in histiocytic necrotizing lymphadenitis

To investigate the mode of presentation, cytologic features of the plasmacytoid dendritic cells (pDC), and the expression of CD123 and its significance in Kikuchi's disease. CD123 expression was evaluated by EliVision immunohistochemical staining in formalin-fixed and paraffin-embedded tissues from 30 cases of Kikuchi's disease, 5 cases of T cell lymphoma, 10 cases of reactive lymphoid hyperplasia and 10 cases of chronic tonsillitis. Clusters of CD123 positive PDC were observed in Kikuchi's disease (28 of 30 cases, 93.3%) and the staining intensity was more prominent in the PDC at the periphery of the lesion and around the high endothelial venule-like vessels. CD123 showed three staining patterns: membranous (10 cases, 33.3%), cytoplasmic (10 cases, 33.3%), and membranous and cytoplasmic (8 cases, 26.7%). In the control group, CD123 showed cytoplasmic staining in reactive hyperplasia and chronic tonsillitis. Regarding the staining intensity, 12 of 28 cases (42.9%) were 3+ for CD123, 8 of 28 cases (28.6%) were 2+, and 8 of 28 cases (28.6%) were 1+. In contrast, PDC clusters with 1+ staining intensity were observed in 1 of 10 cases of reactive lymphoid hyperplasia; 2 of 10 chronic tonsillitis diseases; and much less in T cell lymphoma. Large cluster of PDC is detected in both proliferative and necrotizing types of Kikuchi's disease, making this a useful adjunctive diagnostic marker.

Optimizing the efficacy of targeted phototherapy by marking early vitiligo lesions after visualizing under a Wood's lamp

SOLUTION We have been practicing a simple addition to the technique of targeted phototherapy where the vitiligo lesions are first visualized under a Wood’s lamp (Fig 1) and then marked with a skin marker. The targeted ultraviolet B light phototherapy session is started after the area is marked (Lumera; Daavlin, Bryan, OH). This method ensures that early vitiligo lesions, especially those in the periphery of active lesions, are targeted early, thereby increasing the efficacy of the procedure.

Stereotactic radiosurgery for patients with multiple brain metastases (JLGK0901): a multi-institutional prospective observational study

We aimed to examine whether stereotactic radiosurgery without whole-brain radiotherapy (WBRT) as the initial treatment for patients with five to ten brain metastases is non-inferior to that for patients with two to four brain metastases in terms of overall survival. This prospective observational study enrolled patients with one to ten newly diagnosed brain metastases (largest tumour <10 mL in volume and <3 cm in longest diameter; total cumulative volume ≤15 mL) and a Karnofsky performance status score of 70 or higher from 23 facilities in Japan. Standard stereotactic radiosurgery procedures were used in all patients; tumour volumes smaller than 4 mL were irradiated with 22 Gy at the lesion periphery and those that were 4-10 mL with 20 Gy. The primary endpoint was overall survival, for which the non-inferiority margin for the comparison of outcomes in patients with two to four brain metastases with those of patients with five to ten brain metastases was set as the value of the upper 95% CI for a hazard ratio (HR) of 1·30, and all data were analysed by intention to treat. The study was finalised on Dec 31, 2012, for analysis of the primary endpoint; however, monitoring of stereotactic radiosurgery-induced complications and neurocognitive function assessment will continue for the censored subset until the end of 2014. This study is registered with the University Medical Information Network Clinical Trial Registry, number 000001812. We enrolled 1194 eligible patients between March 1, 2009, and Feb 15, 2012. Median overall survival after stereotactic radiosurgery was 13·9 months [95% CI 12·0-15·6] in the 455 patients with one tumour, 10·8 months [9·4-12·4] in the 531 patients with two to four tumours, and 10·8 months [9·1-12·7] in the 208 patients with five to ten tumours. Overall survival did not differ between the patients with two to four tumours and those with five to ten (HR 0·97, 95% CI 0·81-1·18 [less than non-inferiority margin], p=0·78; pnon-inferiority<0·0001). Stereotactic radiosurgery-induced adverse events occurred in 101 (8%) patients; nine (2%) patients with one tumour had one or more grade 3-4 event compared with 13 (2%) patients with two to four tumours and six (3%) patients with five to ten tumours. The proportion of patients who had one or more treatment-related adverse event of any grade did not differ significantly between the two groups of patients with multiple tumours (50 [9%] patients with two to four tumours vs 18 [9%] with five to ten; p=0·89). Four patients died, mainly of complications relating to stereotactic radiosurgery (two with one tumour and one each in the other two groups). Our results suggest that stereotactic radiosurgery without WBRT in patients with five to ten brain metastases is non-inferior to that in patients with two to four brain metastases. Considering the minimal invasiveness of stereotactic radiosurgery and the fewer side-effects than with WBRT, stereotactic radiosurgery might be a suitable alternative for patients with up to ten brain metastases. Japan Brain Foundation.

Collagen remodeling and peripheral immune cell recruitment characterizes the cutaneous Langerhans cell histiocytosis microenvironment

Langerhans cell histiocytosis (LCH) is a rare and potentially fatal disorder of unknown etiology arising from the accumulation of epidermal Langerhans-like cells in bone, skin, or other tissues. Tissue damage and morbidity results from lesional cytokine release, and we sought to investigate the LCH microenvironment using a combination of histological stains and immunohistochemistry. CD1a immunoreactivity was used to identify lesional cells in archival paraffin-embedded samples of cutaneous LCH. A combined Orcein and Giemsa stain identified immune cells in general (particularly granulocytes and mast cells) and extracellular matrix (particularly elastic fibers), while CD3 and CD68 staining identified T cells and macrophages, respectively. Collagen synthesis was investigated with Herovici staining, which discriminates newly synthesized from mature collagen, while cross-polar microscopy of picrosirius-stained sections identified changes in matrix organization. Immune cells were consistently identified at the periphery of cutaneous LCH lesions. We quantified an increased number of thickened and disorganized elastic fibers surrounding lesions and an absence of elastic fibers within lesions. Furthermore, lesions exhibited a decrease in mature collagen fibers and a loss of supporting collagen matrix within lesions and compromised collagen integrity in adjacent tissue. Cutaneous LCH lesions are associated with the peripheral recruitment of a variety of immune cells and are consistently characterized by localized elastosis, collagen damage, and remodeling.

Role of Perfusion CT Differentiating Hemangiomas from Malignant Hepatic Lesions.

Objective:The purpose of the study was to determine the role of computed tomography (CT) perfusion in differentiating hemangiomas from malignant hepatic lesions.Materials and Methods:This study was approved by the institutional review board. All the patients provided informed consent. CT perfusion was performed with 64 multidetector CT (MDCT) scanner on 45 patients including 27 cases of metastasis, 9 cases of hepatocellular carcinoma (HCC), and 9 cases of hemangiomas. A 14 cm span of the liver was covered during the perfusion study. Data was analyzed to calculate blood flow (BF), blood volume (BV), permeability surface area product (PS), mean transit time (MTT), hepatic arterial fraction (HAF), and induced residue fraction time of onset (IRFTO). CT perfusion parameters at the periphery of lesions and background liver parenchyma were compared.Results:Significant changes were observed in the perfusion parameters at the periphery of different lesions. Of all the perfusion parameters BF, HAF, and IRFTO showed most significant changes. In our study we found: BF of more than 400 ml/100 g/min at the periphery of the hemangiomas showed sensitivity of 88.9%, specificity of 83.3%, positive predictive value (PPV) of 57.1%, and negative predictive value (NPV) of 96.7% in differentiating hemangiomas from hepatic malignancy; HAF of more than 60% at the periphery of hemangiomas showed sensitivity of 77.8%, specificity of 86.1%, PPV of 58.3% and NPV of 93.9% in differentiating hemangiomas from hepatic malignancy; IRFTO of more than 3 s at the periphery of hemangiomas showed sensitivity of 77.8%, specificity of 86.1%, PPV of 58.3%, and NPV of 93.9% in differentiating hemangiomas from hepatic malignancy.Conclusion:Perfusion CT is a helpful tool in differentiating hemangiomas from hepatic malignancy by its ability to determine changes in perfusion parameters of the lesions.

Multifocal testicular capillary hemangioma

Capillary hemangioma of testis is extremely rare. Less than 20 cases have been reported and all of these were single lesion. To our knowledge this is the first case report of a multifocal testicular capillary hemangioma. An elderly patient presented with pain in the right testis. Ultrasound showed a mass in the upper pole with increased vascularity at the periphery of lesion. The left testicle and testicular tumor markers were normal. Radical orchidectomy was performed that showed a well-defined cystic area within the testis. Histology showed an organizing hematoma in the cystic area with benign capillary proliferations in the periphery. There were multiple well-demarcated foci of benign capillary proliferations in the adjacent testicular parenchyma. These proliferative lesions were positive for CD34 and factor VIII. This case was diagnosed as multifocal capillary hemangioma. A review of color Doppler imaging of the right testis showed an area of hypervascularity inferior to the mass lesion that corresponded with multifocal capillary hemangiomas on histology. Vascular neoplasms of testis are rare and mainly reported in young adults. Preoperative imaging and frozen sections are vital for appropriate management of these patients. Conservative management or incomplete excision may cause recurrence or hemorrhage but malignant transformation of these lesions has not been reported. This case of multifocal capillary hemangioma in the testis emphasizes the need for an astute radiological examination with frozen section for the appropriate management of patients with a clinically suspicious testicular mass.

Could ER Stress Be A Major Link Between Oxidative Stress And Autoimmunity In Vitiligo?

Vitiligo is an acquired pigmentary disorder characterized by areas of depigmented skin resulting from loss of epidermal melanocytes. The cause of the destruction of epidermal melanocytes is complex and not yet fully understood. However, there are several hypotheses related to biochemical, neural and genetic aspects as well as oxidative stress and autoimmune mechanisms proposed to understand this disorder. Oxidative stress has a role in vitiligo onset, while autoimmunity contributes to disease progression. In this review, we discuss the mechanisms that link triggering factors with the disease progression. Oxidative stress causes disruption in redox potentials that extend to the Endoplasmic Reticulum (ER), causing accumulation of misfolded proteins, which activates the Unfolded Protein Response (UPR). Melanocytes at the periphery of vitiligo lesions show dilation of the ER. Following exposure to various triggers of vitiligo melanocytes produce cytokines that activate immune response. These studies expand our understanding of the possible underlying mechanisms of melanocyte loss in vitiligo pathogenesis highlighting the possible mechanisms linking ER stress to oxidative stress and autoimmunity.

Effective treatment of congenital melanocytic nevus and nevus sebaceous using the pinhole method with the erbium-doped yttrium aluminium garnet laser.

Dear Editor: Congenital melanocytic nevus (CMN) is a melanocytic nevus that is either present at birth or appears during the latter stages of infancy1. Nevus sebaceous has been described as the hamartomatous locus of an embryologically defective pilosebaceous unit2. Here, we describe how we used the pinhole technique with an erbium-doped yttrium aluminium garnet (erbium : YAG) laser to treat nevi lesions in different patients. A 25-year-old male was diagnosed with CMN on his nose at birth (Fig. 1A). Emla cream (AstraZeneca AB, Sodertalje, Sweden) was applied to the treatment area prior to starting the procedure. The erbium : YAG laser (Action Er : YAG; Lutronic Corporation, Goyang, Korea) was used in continuous-wave mode, with 1-mm spot size and output power of 0.2 mJ/cm2. Multiple small holes measuring 1 mm in diameter were made on the periphery of the CMN lesion. A hydrocolloid dressing (DuoDERM Extra Thin CGF dressing; ConvaTec, ER Squibb & Sons, Princeton, NJ, USA) was applied to the lesions for ≥2 weeks after treatment. After performing 5 laser sessions at 4-week intervals, the cutaneous lesions resolved. There was no evidence of recurrence at the 1-year follow-up examination (Fig. 1B). Fig. 1 Congenital melanocytic nevus (CMN) in a 25-year-old man. (A) Before treatment: a solitary, 1-cm brown patch is visible on the right alar area of the nose. (B) One year after the final treatment session, the skin lesions resolved with minimal scarring. ... A 5-month-old infant was referred to our outpatient unit for CMN on the forearm (Fig. 1C). We performed the pinhole method of treatment using the erbium : YAG laser (Lutronic) in continuous-wave mode, with 1-mm spot size and output power of 0.2 mJ/cm2 (Fig. 1D). After receiving 6 treatment sessions at 2-month intervals, slight superficial pigmentation persisted (Fig. 1E). A 40-year-old woman presented with nevus sebaceous lesions on the right side of her forehead (Fig. 2A). We performed the pinhole method of treatment using the erbium : YAG laser (Lutronic) in continuous-wave mode, with 1-mm spot size and output power of 0.2 mJ/cm2. After receiving 5 treatment sessions at 4-week intervals, the patient showed marked improvement with only mild hypopigmentation (Fig. 2B). No recurrence was noted at the 6-month follow-up examination (Fig. 2C). Fig. 2 Nevus sebaceous lesions in a 40-year-old woman. (A) Before treatment, two 2.5-cm, well-demarcated, yellow, verrucous plaques were visible on the right side of the forehead. (B) Deep line- and doughnut-shaped holes were created to remove the nevus sebaceus ... Herein, we described how nevi lesions were effectively treated using the pinhole method. This simple method involves making multiple small holes at 2~5-mm intervals using a CO2 laser that penetrates the epidermis and deeper dermis3. The holes produced using the pinhole method are surrounded by untreated areas that promote rapid epidermal repair, thereby reducing the downtime and likelihood of side effects4. The other advantages of performing the pinhole method with ablative lasers are that this laser is easy to use and inexpensive, as ablative lasers are usually available in most dermatology clinics. To the best of our knowledge, this is the first report on the application of the pinhole method for the treatment of CMN and nevus sebaceous, so it is difficult to compare the advantages and disadvantages of the pinhole method with those of conventional treatment methods. Therefore, it is necessary to conduct further studies with a large sample to confirm the effectiveness and safety of the pinhole method. In conclusion, we suggest using the pinhole method with erbium : YAG laser as an alternative therapy for treating nevi lesions.

In Vivo Theranostics at the Peri-Infarct Region in Cerebral Ischemia

The use of theranostics in neurosciences has been rare to date because of the limitations imposed on the free delivery of substances to the brain by the blood-brain barrier. Here we report the development of a theranostic system for the treatment of stroke, a leading cause of death and disability in developed countries. We first performed a series of proteomic, immunoblotting and immunohistological studies to characterize the expression of molecular biomarkers for the so-called peri-infarct tissue, a key region of the brain for stroke treatment. We confirmed that the HSP72 protein is a suitable biomarker for the peri-infarct region, as it is selectively expressed by at-risk tissue for up to 7 days following cerebral ischemia. We also describe the development of anti-HSP72 vectorized stealth immunoliposomes containing imaging probes to make them traceable by conventional imaging techniques (fluorescence and MRI) that were used to encapsulate a therapeutic agent (citicoline) for the treatment of cerebral ischemia. We tested the molecular recognition capabilities of these nano-platforms in vitro together with their diagnostic and therapeutic properties in vivo, in an animal model of cerebral ischemia. Using MRI, we found that 80% of vectorized liposomes were located on the periphery of the ischemic lesion, and animals treated with citicoline encapsulated on these liposomes presented lesion volumes up to 30% smaller than animals treated with free (non-encapsulated) drugs. Our results show the potential of nanotechnology for the development of effective tools for the treatment of neurological diseases.

101 Vitamin D3 cryosensitization in androgen insensitive prostate cancer

Cryoablation is increasingly utilized as both a primary and salvage treatment option for prostate cancer (PC). The dynamic nature of a cryolesion provides a temperature gradient which correlates with complete cell death at the core of the iceball to a heterogeneous zone of live cells at the elevated temperatures near the periphery. The utilization of molecular adjuvants aims to achieve an increased ablative zone at these warmer isotherms. Previous studies have confirmed the synergistic effects of chemotherapeutic drugs in combination with freezing temperatures associated with the periphery of a cryogenic lesion (−10 °C through −25 °C). More recently we have utilized the active metabolite of vitamin D3 (calcitriol) as a cryosensitizing agent in numerous in vitro models. In both murine and human studies we found that calcitriol pre-treatment yielded a significant increase in cell death over freezing alone. Additionally, both androgen sensitive (AS) and androgen insensitive (AI) human PC models respond to calcitriol cryosensitization. Given the difficulty in treating AIPC clinically, we further investigated the sensitization capabilities of calcitriol in this model. Specifically, in the AIPC line PC-3, 24 h calcitriol pre-treatment yielded a 36% increase in cell death at the −15 °C isotherm compared to temperature-matched controls (60% viable vs. 96%, respectively). In addition to 2D in vitro modeling, studies were also conducted using a novel 3D prostate tissue engineered model (pTEM) system. PC-3 cells grown in this 3D matrix were frozen using a novel supercritical nitrogen (SCN) device in the presence or absence of calcitriol as a cryosensitizer and the effectiveness of calcitriol pre-treatment was evaluated. Results indicate that the zone of cellular ablation was increased by 20% with the addition of calcitriol as compared to non-treated frozen samples, illustrating an increase in cell death while simultaneously reducing the need for an enlarged positive freeze margin. These studies demonstrate the in vitro effectiveness of a combinatorial approach to AIPC treatment utilizing calcitriol and cryotherapy and thus illustrate the potential for future clinical applications. Source of funding: CPSI Biotech. Conflict of interest: None declared. ksantucci@cpsibiotech.com

Lesion-Induced Alterations in Astrocyte Glutamate Transporter Expression and Function in the Hippocampus

Astrocytes express the sodium-dependent glutamate transporters GLAST and GLT-1, which are critical to maintain low extracellular glutamate concentrations. Here, we analyzed changes in their expression and function following a mechanical lesion in the CA1 area of organotypic hippocampal slices. 6-7 days after lesion, a glial scar had formed along the injury site, containing strongly activated astrocytes with increased GFAP and S100β immunoreactivity, enlarged somata, and reduced capability for uptake of SR101. Astrocytes in the scar's periphery were swollen as well, but showed only moderate upregulation of GFAP and S100β and efficiently took up SR101. In the scar, clusters of GLT-1 and GLAST immunoreactivity colocalized with GFAP-positive fibers. Apart from these, GLT-1 immunoreactivity declined with increasing distance from the scar, whereas GLAST expression appeared largely uniform. Sodium imaging in reactive astrocytes indicated that glutamate uptake was strongly reduced in the scar but maintained in the periphery. Our results thus show that moderately reactive astrocytes in the lesion periphery maintain overall glutamate transporter expression and function. Strongly reactive astrocytes in the scar, however, display clusters of GLAST and GLT-1 immunoreactivity together with reduced glutamate transport activity. This reduction might contribute to increased extracellular glutamate concentrations and promote excitotoxic cell damage at the lesion site.

First Report of Leaf Spot of Phoenix theophrasti Caused by Paraconiothyrium variabile in Greece.

In the spring of 2011, a severe leaf spot disease of Phoenix theophrasti was observed in the vicinity of Heraklion (Crete), Greece. Initial symptoms were small, round-ovoid spots of varying shades of brown on the leaves, later being transformed into oblong streaks (average dimensions 7.3 ± 1.0 × 3.3 ± 0.5 mm), surrounded by dark brown rings. As the disease progressed, the expanding streaks often coalesced to form enlarged necrotic lesions. Similar symptoms were also detected on petioles and leaf bases. Extended spotting and blighting occasionally resulted in leaf death. A filamentous fungus was consistently isolated onto potato dextrose agar plates from the periphery of the characteristic lesions, with cultures invariably producing brick to cinnamon colonies with sparse aerial mycelium, subglobose and dark brown superficial pycnidial conidiomata on pine needles, 1- to 3-celled hyaline conidiophores, and hyaline, subcylindrical to ellipsoidal, 1-celled, smooth- and thin-walled conidia, with average dimensions of 3.5 ± 0.6 × 1.7 ± 0.4 μm (n = 100). Total DNA of two isolates was extracted and used for PCR amplification and sequencing of the ITS1-5.8S-ITS2 region, together with parts of the flanking 18S and 28S rRNA genes (4). Both sequences (GenBank Accession Nos. JX456476 and JX456477) were 100% identical to deposited Paraconiothyrium variabile ITS sequences (EU295640 to 48, JN983440 and 41, and JF934920), and were clustered together as a single group with these sequences with good support by phylogenetic analysis that included representatives of the relative P. brasiliense and P. africanum species. Based on the morphological, molecular, and phylogenetic analyses, the pathogen was identified as P. variabile Riccioni, Damm, Verkley & Crous (2). To prove pathogenicity, 10 P. theophrasti 2-year-old seedlings were sprayed with a conidial suspension of the fungus (107 conidia ml-1, 10 ml per plant), while five additional control plants were treated with sterile distilled water. All plants were maintained in the greenhouse at 15 ± 5°C, with 90% humidity. Characteristic leaf spots were evident 4 weeks post inoculation on the older leaves, and P. variabile was consistently reisolated from all inoculated plants. No symptoms were observed on control plants. Paraconiothyrium variabile has been isolated from various woody host plants such as Prunus persica, P. salicina, and Malus sp. in South Africa (1,2), Actinidia chinensis and A. deliciosa in Italy (2), Laurus nobilis in Turkey (2), and Salix matsudana in China (3). To our knowledge, this is the first report of P. variabile naturally infecting and causing a leaf spot disease on a palm species. Palms are extensively used as ornamentals throughout Greece and the occurrence of P. variabile can potentially result in economic loss to the local ornamental industry.

Atypical idiopathic inflammatory demyelinating lesions (IIDL): Conventional and diffusion-weighted MR imaging (DWI) findings in 42 cases

IntroductionThe purpose of this study was to evaluate MR imaging characteristics with conventional and advanced MR imaging techniques in patients with IIDL. MethodsMR images of the brain in 42 patients (20 male, 22 female) with suspected or known multiple sclerosis (MS) from four institutions were retrospectively analyzed. Lesions were classified into five different subtypes: (1) ring-like lesions; (2) Balo-like lesions; (3) diffuse infiltrating lesions; (4) megacystic lesions; and (5) unclassified lesions.The location, size, margins, and signal intensities on T1WI, T2WI, and diffusion-weighted images (DWI), and the ADC values/ratios for all lesions, as well as the contrast enhancement pattern, and the presence of edema, were recorded. ResultsThere were 30 ring-like, 10 Balo-like, 3 megacystic-like and 16 diffuse infiltrating-like lesions were detected. Three lesions were categorized as unclassified lesions.Of the 30 ring-like lesions, 23 were hypointense centrally with a hyperintense rim. The mean ADC, measured centrally, was 1.50±0.41×10−3mm2/s. The mean ADC in the non-enhancing layers of the Balo-like lesions was 2.29±0.17×10−3mm2/s, and the mean ADC in enhancing layers was 1.03±0.30×10−3mm2/s. Megacystic lesions had a mean ADC of 2.14±0.26×10−3mm2/s. Peripheral strong enhancement with high signal on DWI was present in all diffuse infiltrating lesions. Unclassified lesions showed a mean ADC of 1.43±0.13mm2/s. ConclusionRestriction of diffusion will be seen in the outer layers of active inflammation/demyelination in Balo-like lesions, in the enhancing part of ring-like lesions, and at the periphery of infiltrative-type lesions.