Synthesis and preclinical evaluation of BOLD-100 radiolabeled with ruthenium-97 and ruthenium-103.

BOLD-100 (formerly IT-139, KP1339), a well-established chemotherapeutic agent, is currently being investigated in clinical trials for the treatment of gastric, pancreatic, colorectal, and bile duct cancer. Despite numerous studies, the exact mode of action is still the subject of discussions. Radiolabeled BOLD-100 could be a powerful tool to clarify pharmacokinetic pathways of the compound and to predict therapy responses in patients using nuclear molecular imaging prior to the therapy. In this study, the radiosyntheses of carrier-added (c.a.) [97/103Ru]BOLD-100 were performed with the two ruthenium isotopes ruthenium-103 (103Ru; ß-, γ) and ruthenium-97 (97Ru; EC, γ), of which in particular the latter isotope is suitable for imaging by single-photon emission computed tomography (SPECT). To identify the best tumor-to-background ratio for diagnostic imaging, biodistribution studies were performed with two different injected doses of c.a. [103Ru]BOLD-100 (3 and 30 mg kg-1) in Balb/c mice bearing CT26 allografts over a time period of 72 h. Additionally, ex vivo autoradiography of the tumors (24 h p.i.) was conducted. Our results indicate that the higher injected dose (30 mg kg-1) leads to more unspecific accumulation of the compound in non-targeted tissue, which is likely due to an overload of the albumin transport system. It was also shown that lower amounts of injected c.a. [103Ru]BOLD-100 resulted in a relatively higher tumor uptake and, therefore, a better tumor-to-background ratio, which are encouraging results for future imaging studies using c.a. [97Ru]BOLD-100.

Histopathological examination of characteristic brain MRI findings in acute hyperammonemic encephalopathy: A case report and review of the literature.

INTRODUCTION: Acute hyperammonemic encephalopathy is associated with distinct brain MRI findings, namely, hyperintensity in T2-weighted sequences as well as restricted diffusion in diffusion-weighted imaging with accentuation in the insular cortex and cingulate gyrus. The pathophysiology and the histopathological correlates of these characteristic MRI findings are largely unknown. CASE REPORT: We present a 57-year-old male with a history of chronic alcohol abuse, liver cirrhosis and portal hypertension, and a clinical syndrome (variceal bleeding, depression of consciousness, seizures), elevated plasma ammonia levels, and characteristic brain MRI abnormalities suggestive of acute hyperammonemic encephalopathy. A postmortem histopathological examination revealed extensive hypoxic ischemic encephalopathy without evidence for metabolic encephalopathy. No episodes of prolonged cerebral hypoxemia were documented throughout the course of the disease. We conducted a review of the literature, which exhibited no reports of hyperammonemic encephalopathy in association with characteristic brain MRI findings and a consecutive histopathological examination. CONCLUSION: This is the first report of a patient with acute hyperammonemic encephalopathy together with characteristic brain MRI findings and a histopathological correlation. Although characteristic MRI findings of acute hyperammonemic encephalopathy were present, a histopathological examination revealed only hypoxic pathology without signs of metabolic encephalopathy.

CT-guided piriformis muscle injection for the treatment of piriformis syndrome.

AIM: Piriformis syndrome is a rare neuromuscular disorder that occurs when the piriformis muscle compresses or irritates the sciatic nerve. The treatment of piriformis syndrome includes injections into the piriformis muscle around the sciatic nerve. These invasive approaches have been used with various techniques to increase the safety of the procedure. Computed tomography (CT)-guided injection of the piriformis muscle and the clinical outcome of the patients are discussed. MATERIAL AND METHODS: The authors presented 10 consecutive patients that underwent CT-guided piriformis injection between March and December 2007. Three patients had a history of a severe fall on the buttocks, one had gluteal abscess formation following deep intramuscular injection, and another one had a habit of prolonged sitting on the carpet. Etiology was not identified in the other patients. Main complaints of the patients were pain and numbness in the legs. Hypesthesia was the major neurological finding. Magnetic resonance imaging (MRI) and electromyography (EMG) were performed in all patients. RESULTS: Nine patients had full and sustained recovery of their symptoms after piriformis injection. Only the patient who had gluteal abscess formation following deep intramuscular injection showed moderate improvement. Another patient was operated on in the 6th month after piriformis injection due to an extruded disc herniation. CONCLUSION: CT-guided piriformis injection is a safe and effective method in the treatment of piriformis syndrome.

Development of peptide aptamer microarrays for detection of HPV16 oncoproteins in cell extracts.

Protein microarrays represent an emerging technology that promises to facilitate high-throughput proteomics. The major goal of this technology is to employ peptides, full-length proteins, antibodies, and small molecules to simultaneously screen thousands of targets for potential protein-protein interactions or modifications of the proteome. This article describes the performance of a set of peptide aptamers specific for the human papillomavirus (HPV) type 16 oncoproteins E6 and E7 in a microarray format. E6 and E7 peptide aptamer microarrays were probed with fluorescence-labeled lysates generated from HPV-infected cervical keratinocytes expressing both E6 and E7 oncoproteins. Peptide aptamer microarrays are shown to detect low levels of E6 and E7 proteins. Peptide aptamers specific for cellular proteins included on these microarrays suggested that expression of CDK2, CDK4, and BCL-6 may be affected by HPV infection and genome integration. We conclude that peptide aptamer microarrays represent a promising tool for proteomics and may be of value in biological and clinical investigations of cervical carcinogenesis.

Electron microscopic examination of the myelinated axons of corpus callosum in perfused young and old rats.

In this study, the myelinated axons of parts of the corpus callosums of young and old rats were examined under the electron microscope and a grading system was performed for quantitating the ultrastructural pathological changes of these axons. Except the old splenium group, the only ultrastructural pathological change, observed in the myelinated axons was the separation in myelin configuration. In addition to this finding, in the old splenium group, in some of the myelinated axons, an interruption was observed in the myelin configuration. Additionally, these ultrastructural pathological findings were present in the larger sized myelinated axons of the corpus callosum.

Light and electron microscopic examination of human subungual tissue. Glomus and lamellated bodies.

OBJECTIVE: There is only limited data related to the subungual glomus body. We therefore studied the structure of this organ, aiming to obtain further evidence. Additionally, we encountered undefined receptor like structures in close association with these glomus cells, named them as lamellated bodies and examined both of the structures at light and electron microscopic levels. METHODS: This study was carried out at the Faculty of Medicine, Hacettepe University, Ankara, Turkey, during the time period May 2001 to March 2002. In this study, the subungual tissues of 4 patients were examined. RESULTS: Within subungual tissue, 2 groups of morphologically significant structures were determined by light microscopy. The first structure was described as glomus body. It was characterized as an encapsulated structure, rich in rounded clear cells filling its central compartment. The latter structure having a lamellated appearance was described as lamellated body. In the electron microscopic examination, lamellated bodies were characterized by central filament rich large cells and surrounding cytoplasmic processes of ensheathing cells, some of which were vacuolated. Glomus bodies were surrounded by a capsule and centrally located numerous rounded cells which reflected the structural features of an active cell. CONCLUSION: The lamellated bodies are very unusual structures and they are not found in any other part of the body. The structural organization of the ensheathing cells in the lamellated bodies greatly resembles many skin associated receptors. Therefore, we planned future studies by using immunohistochemistry, to reveal nervous elements for structural contribution.

Right internal iliac vein joining the left common iliac vein: case report demonstrated by CT angiography.

An unusual variation of the iliac veins was detected by computed tomography (CT) angiography in a 35-year-old man. In coronal CT reconstructions, it was shown that the right internal iliac vein of this patient crossed to the left side and drained to the left common iliac vein. This variation is important in retroperitoneal, laparoscopic and orthopedic surgery. We present the CT findings and discuss the embryological origin of this unusual congenital anomaly.

Double tendons at the distal attachment of the extensor hallucis longus muscle.

The distal attachments of the extensor hallucis longus (EHL) tendons in 47 amputated legs and in eight cadavers were examined. The EHL had two tendons in 34 of the amputated legs and bilaterally in five cadavers. The lateral tendon was inserted to the middle of the dorsal aspect of the base of the distal phalanx of the hallux and the medial tendon to the medial side of the insertion of the lateral tendon. The length and thickness of these two tendons were measured and compared in order to obtain data for using these tendons in tendon repair and hallux varus corrections by autogenous tendon transfer surgery. Additionally, on the right foot of one of the cadavers, it was observed that the extensor hallucis brevis tendon united with the lateral tendon of the EHL. We recommend that foot-ankle surgeons be aware of the various accessory EHL tendons and their potential use in problematic cases.

Peptide aptamers targeting the hepatitis B virus core protein: a new class of molecules with antiviral activity.

A substantial proportion of the worldwide liver cancer incidence is associated with chronic hepatitis B virus (HBV) infection. The therapeutic management of HBV infections is still problematic and novel antiviral strategies are urgently required. Using the peptide aptamer screening system, we aimed to isolate new molecules, which can block viral replication by interfering with capsid formation. Eight peptide aptamers were isolated from a randomized expression library, which specifically bound to the HBV core protein under intracellular conditions. One of them, named C1-1, efficiently inhibited viral capsid formation and, consequently, HBV replication and virion production. Hence, C1-1 is a novel model compound for inhibiting HBV replication by blocking capsid formation and provides a new basis for the development of therapeutic molecules with specific antiviral potential against HBV infections.

Peptide aptamers: new tools to study protein interactions.

The ability to specifically interfere with the function of proteins of pathological significance has been a goal for molecular medicine for many years. Peptide aptamers comprise a new class of molecules, with a peptide moiety of randomized sequence, which are selected for their ability to bind to a given target protein under intracellular conditions. They have the potential to inhibit the biochemical activities of a target protein, can delineate the interactions of the target protein in regulatory networks, and identify novel therapeutic targets. Peptide aptamers represent a new basis for drug design and protein therapy, with implications for basic and applied research, for a broad variety of different types of diseases.

p53 mutations are rare events in recurrent cervical cancer.

Mutations of the p53 gene have been shown to be associated with aggressive growth behavior and increased recurrence rates for certain tumors. Primary cervical cancers contain oncogenic human papillomaviruses (HPV) in more than 90% of cases and usually possess wild-type p53 alleles. Cervical cancer cells contain detectable levels of functional p53 protein despite of the expression of the HPV E6 protein, which can induce p53 degradation. Thus, inactivation of p53 by somatic mutation should have functional consequences in HPV-positive cancers. We investigated whether p53 mutations play a role in the recurrence of the disease by analyzing p53 status in 18 biopsy specimens from recurrent cervical cancers. Only one of these (5.6%) contained a p53 mutation, as assessed by a sensitive yeast functional assay that detects mutations of the p53 mRNA between codons 52 and 364. These results indicate that p53 mutations are rare events in recurrent cervical carcinomas, and that somatic mutations of p53 do not provide cervical cancer cells with a selective growth advantage for recurrence.

Expression pattern of AP-2 transcription factors in cervical cancer cells and analysis of their influence on human papillomavirus oncogene transcription.

The AP-2 family of transcription factors consists of three known members, namely AP-2alpha, AP-2beta, and AP-2gamma. In experimental systems AP-2 factors possess tumor suppressor-like activities, and alterations in the AP-2 expression pattern have been described for some tumor entities. In addition, AP-2 has been implicated in the transcriptional control of human papillomaviruses (HPVs). We investigated here the expression pattern of AP-2alpha, AP-2beta, and AP-2gamma, as well as that of the cellular AP-2 target gene c-erbB-2, in a series of cervical cancer cell lines. In addition, we analyzed the influence of AP-2 factors on the activity of the HPV16 and HPV18 E6/E7 oncogene promoter. We found that, with the exception of HPV-negative C33A cells, all investigated cervical cancer cell lines expressed all three AP-2 family members, although at varying levels. No linear correlation between AP-2 and c-erbB-2 levels was observed. Although AP-2alpha, AP-2beta, and AP-2gamma can activate the c-erbB-2 promoter in reporter gene assays, they do not stimulate the HPV16 or HPV18 E6/E7 promoter. These results indicate that, although a rare event, loss of AP-2 expression occurs in cervical cancer cells. Moreover, AP-2alpha, AP-2beta, and AP-2gamma are neither sufficient nor required to activate the viral E6/E7 promoter.

Do consumers know how their health plan works?

Expanding consumer choice of plans is beneficial only to the extent that consumers make informed choices. Using data from the 1996-97 Community Tracking Study (CTS), this study compares consumers' responses on four key attributes of their health plan with information provided directly by the plan. Plan attributes relate to choice of providers and access to specialists. Although the accuracy of reporting some individual attributes was fairly high, fewer than one-third of consumers accurately reported all four health plan attributes. In general, consumers tended to overreport plan restrictions, especially the need for approval to see specialists.

Public opinion on alcohol policies in the United States: results from a national survey.

We surveyed the U.S. non-institutionalized population age 18+ on opinions regarding 23 alcohol control policies (N = 7,021). The cooperation rate among contacted households was 70% and the overall response rate was 54%. Results showed high levels of public support for most alcohol control policies. Over 80% support restrictions on alcohol use in public places, such as parks, beaches, concert venues, and on college campuses. Eighty-two percent support increased alcohol taxes, provided the funds are used for treatment or prevention programs. Over 60% support alcohol advertising and promotion restrictions, such as banning billboard advertising, banning promotion at sporting events, or banning liquor and beer advertising on television. Multivariate regression analyses indicated significant relationships between alcohol policy opinions and a variety of sociodemographic, political orientation, and behavioral measures. However, the absolute differences in alcohol policy support across groups is small. There is a strong base of support for alcohol control policies in the U.S., and such support is found among whites and ethnics of color, young and old, rich and poor, and conservatives, moderates, and liberals.

Induction of apoptosis in human papillomaviruspositive cancer cells by peptide aptamers targeting the viral E6 oncoprotein.

Certain types of human papillomaviruses (HPVs) are closely linked to the development of human cancers. Herein, it is shown that intracellular targeting of the HPV16 E6 oncoprotein by E6-binding peptide aptamers resulted in the apoptotic elimination of HPV16-positive cancer cells, whereas HPV-negative cells were not affected. These results provide direct experimental evidence that the HPV E6 oncoprotein has antiapoptotic activity in HPV-positive tumor cells that is required for their survival. The E6-targeting molecules identified herein have implications for the development of therapeutic strategies for the treatment of HPV-associated dysplasias and cancers.

Genetic complementation to non-tumorigenicity in cervical-carcinoma cells correlates with alterations in AP-1 composition.

The transcription factor AP-1 represents a central key element in the expression of human pathogenic papillomaviruses (HPV). We here propose a novel role for AP-1 as an essential component of an intracellular surveillance mechanism negatively controlling the proliferation of HPV-positive cells under in vivo conditions. The dissection of AP-1 composition in cervical-carcinoma cells revealed an inverse relationship between the Fos-related antigen Fra-1 and the tumorigenic phenotype. Cervical-carcinoma cell lines were either negative or expressed only low amounts of Fra-1 (jointly with c-Fos) within their AP-1 complexes. Somatic-cell hybridization technique was used to fuse different HPV-positive malignant cell lines. This resulted either in tumorigenic hybrids or in cells in which the malignant phenotype of the parental fusion partners was completely suppressed. The monitoring of AP-1 composition in electrophoretic mobility super-shift assays showed that the amount of Fra-1 was substantially increased within the AP-1 complex of non-malignant cells. In contrast, Fra-1 was even diminished in malignant hybrids, while c-Fos remained expressed. This correlation suggests that the concentration of Fra-1 within the AP-1 transcription complex might be an important marker for predicting the in vivo growth properties of HPV-positive cells.

Growth inhibition of cervical cancer cells by the human retinoic acid receptor beta gene.

Transcription of the retinoic receptor beta (RARbeta) gene is activated in a ligand-dependent manner by the retinoic acid receptor alpha. Reduced RARbeta gene expression and loss of ligand inducibility are frequently observed in human carcinoma cells indicating that such alterations might contribute to carcinogenesis. In this study we have analyzed the influence of RARbeta on cervical cancer cell growth. Transfection of HeLa cells with RARbeta expression plasmids resulted in reduced clonal cell growth in the presence of retinoic acid (RA). RA-induced growth inhibition in HeLa x fibroblast hybrid cells was partially relieved by a dominant-negative RARbeta mutant. HeLa clones stably expressing a RARbeta transgene under control of the human beta-actin promoter [HeLa(RARbeta)] were established and analyzed for transgene-mediated growth alterations in vitro and in vivo. Anchorage-independent growth of the HeLa(RARbeta) lines was indistinguishable from that of control cells in the absence of RA, but strongly impaired after RA treatment. Reduced tumor growth of HeLa(RARbeta) clones was associated with high RARbeta protein levels. Somatic cell fusion experiments revealed that the loss of ligand inducibility of RARbeta gene expression in HeLa cells cannot be complemented by fusion with other cervical cancer cell lines. Our data indicate, firstly, that RARbeta is a negative regulator of tumor cell growth and, secondly, that cancer-associated defects in RARbeta gene expression are caused by stable, non-complementable silencing mechanisms.