Fox parasites in pre-Columbian times: Evidence from the past to understand the current helminth assemblages.

This work aims to increase the information on the entero-parasitism in Holocene carnivores, by examining coprolites found in Patagonia. Molecular analysis was conducted following the Authenticity Criteria to Determine Ancient DNA sequences. The nucleotide sequences showed 99% of identity with the Control Region sequences of Lycalopex culpaeus (culpeo fox). Coprolites were positive for gastrointestinal parasites. The presence of Alaria sp. and Clonorchis sp. represents the first record for pre-Columbian America. The parasitological findings suggest the importance of these carnivores for the dissemination of their own parasites and those to their prey in rockshelters, areas with high re-use of space.

Idiopathic lingual artery aneurysm: CT findings and endovascular therapy. A case report.

SUMMARY: We describe a 65-year-old woman with an asymptomatic idiopathic lingual artery aneurysm which is suspected to be congenital. We review the literature on external carotid artery branch aneurysms, diagnostic evaluation and discuss treatment options for the various types and the specific chosen in the case presented.

1-(3-Chloro-4-fluoro-phen-yl)-5-(2-diazo-acet-yl)-4-phenyl-pyrrolidin-2-one.

In the title compound, C(18)H(13)ClFN(3)O(2), the pyrrolidine ring adopts an envelope conformation and the planar part is rotated by 4.3 (6)° from the plane of the benzene ring and is almost perperdicular both to the diazo-acetyl unit [dihedral angle = 78.93 (7)°] and the phenyl ring [dihedral angle = 86.07 (7)°]. In the crystal, mol-ecules are linked into a three-dimensional framework by C-H⋯O inter-actions. The mol-ecular conformation is stabilized by an intra-molecular C-H⋯O hydrogen bond.

Diethyl 1-(4-methyl-phen-yl)-3-phenyl-5-oxopyrrolidine-2,2-dicarboxyl-ate.

In the title compound, C(23)H(25)NO(5), the lactam ring adopts an envelope conformation and both eth-oxy-carbonyl side chains show an s-cis conformation: one is nearly planar, the dihedral angle between CO(2) and OCH(2)CH(3) groups being 7.95 (14)° and the other is almost orthogonal, the C-O-C-C torsion angle being 85.33 (9)°. Dimers related by inversion symmetry are stabilized by C-H⋯O hydrogen bonds. The crystal structure is consolidated by weak intermolecular C-H⋯O inter-actions. Weak intra-molecular inter-actions of the same kind also occur.

Pediatric Thoraco-lumbar Paraspinal Arteriovenous Fistulas along the Segmental Nerve. Diagnosis and Endovascular Treatment.

SUMMARY: To characterize the clinical presentation, imaging features and endovascular treatment of paraspinal non-vertebral arteriovenous fistulas along the segmental nerve. Retrospective review was performed on the five patients identified in our database covering 1985 to 2003. All patients presented with an incidentally found continuous murmur over the upper paraspinal or parasternal regions before three years old. In four patients, the AV fistula was in the mid-thoracic level and at L3 in one. All AV fistulas were a high-flow single-hole fistula at the neural foramen with venous drainage into paraspinal and epidural veins without intradural reflux. All fistulas were endovascularly occluded in the same session as the diagnostic angiography. The fistula was occluded with detachable coils in one case and with N-butylcyanoacrylate (NBCA) with flow control in four cases. Complete occlusion of the fistula was obtained in all cases and all patients remained neurologically intact at the last follow up (average six years). Non-vertebral paraspinal arteriovenous fistula along the segmental nerve is a specific disease entity seen in children. Embolization is the first choice of treatment for this disease.

COX-2, NSAIDs and human neoplasia. Part I: Colorectal neoplasms.

Cyclooxygenase-2 (COX-2), the inducible cyclooxygenase isozyme involved in the conversion of arachidonic acid (AA) to biologically active prostanoids, has become the subject of intense interest during the last few years. The recent surge of interest stems from seminal studies that correlated elevated expression of COX-2 with tumor induction and progression, and epidemiological studies that correlated reduced risk of developing certain types of cancers with chronic use of non-steroidal anti-inflammatory agents (NSAIDs). Although these observations were first reported with colorectal cancer (CRC), similar findings have subsequently been made with other types of cancers. A wide spectrum of studies continue to be undertaken in both laboratory and clinical settings to elucidate the mechanisms underlying these anti-tumor effects of COX-2 for potential translation into cancer chemoprevention and therapy. The aim of this article is to present a review of COX genes, the prostaglandin-cyclooxygenase relationship, the role of COX-2 in carcinogenesis and the rationale for targeting COX-2 with NSAIDs for cancer chemoprevention. Special emphasis is given to the role of COX-2 expression in the genesis and progression of colorectal neoplasia, and its correlation with other pathological characteristics of CRC. Preliminary observations on COX-2 expression in inflammatory bowel disease (IBD)-related colorectal neoplasia are also presented.

Enhancing quantification of mammalian cell transfections with chloramphenicol acetyltransferase reporter plasmids.

We modified existing techniques to optimize conditions for obtaining quantitative, highly replicable, and sensitive transfections. The processes described may serve as a model for investigators initiating transfection procedures who wish to obtain definitive and quantitative results quickly and efficiently. In our example, we compared specific gene expressions of plasmids with the chloramphenicol acetyltransferase (CAT) reporter. Techniques included measuring CAT activity in transfected mammalian cells, selecting a procedure for extracting plasmids from bacterial cells, evaluating the timing of the transfection, choosing a transfection reagent and the reagent: plasmid DNA ratio, and determining procedures for the extraction of cells.

Inhibitors of prostaglandin synthesis inhibit human prostate tumor cell invasiveness and reduce the release of matrix metalloproteinases.

Eicosanoids modulate the interaction of tumor cells with various host components in cancer metastasis. Their synthesis involves the release of arachidonic acid (AA) from cellular phospholipids by phospholipase A2 (PLA2), followed by metabolism by cyclooxygenases (COXs) and lipooxygenases (LOXs). This study aimed to identify the pathway(s) of AA metabolism that are required for the invasion of prostate tumor cells. DU-145 and PC-3 human prostate cancer cell lines were used to test the effect of inhibitors of PLA2, COX, or LOX on the invasion of prostate tumor cells through Matrigel in vitro using the Boyden chamber assay and fibroblast-conditioned medium as the chemoattractant. We used nontoxic doses that did not inhibit simple cell motility and did not decrease clonogenic survival. All of the inhibitors caused a significant reduction in AA release from treated cells compared with control cells, which indicated that the treatments were biochemically active. Invasion through Matrigel was inhibited by the PLA2 inhibitor 4-bromophenacyl bromide (4-BPB), the general COX inhibitor ibuprofen (IB), and the highly selective COX-2 inhibitor NS398. Inhibition of cell invasiveness by 4-BPB (1.0 microM), IB (10.0 microM), and NS398 (10.0 microM) was reversed by the addition of prostaglandin E2 (PGE2). PGE2 alone, however, did not stimulate invasiveness, which suggests that its production is necessary for rendering the cells invasive-permissive but not sufficient for inducing invasiveness. In contrast, we found no significant inhibition of invasion of prostate tumor cells treated with esculetin (1.0 microM) or nordihydroguiaretic acid (1.0 microM), which are specific inhibitors of LOX. We also tested the effect of 4-BPB, IB, NS398, and esculetin on the secretion of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), as key enzymes in the proteolysis of Matrigel during invasion, using gelatin zymograms and Western blots. Cells that received 4-BPB, IB, or NS398, but not esculetin showed a significant reduction in the levels of proMMP-2, MMP-9, and proMMP-9 in the culture medium. DU-145 cells did not secrete TIMP-1, and the drugs did not alter the secretion of TIMP-2. This work highlights the role played by COX in disturbing the balance between MMPs and TIMPs in prostate cancer cells, and it points to the potential use of COX inibitors, especially COX-2 selective inhibitors, in the prevention and therapy of prostate cancer invasion.

Alternative splicing prevents transferrin secretion during differentiation of a human oligodendrocyte cell line.

Transferrin, the iron-transport protein of vertebrate serum, is synthesized mainly in the liver, from which it is secreted into the blood. Transferrin is also synthesized in oligodendrocytes and is an early marker of their differentiation. We have analyzed the regulation of transferrin expression in HOG cells, a human oligodendrocyte cell line. Transferrin expression was correlated with the appearance of oligodendrocyte differentiation markers when cells were exposed to differentiation medium. In contrast to the protein expressed in hepatocytes or in Sertoli cells, transferrin was secreted by neither HOG cells nor immature rat primary oligodendrocytes in vitro. Moreover, transferrin appears to be localized in the cytosol and not in the secretory compartment, as is expected for secreted proteins. This transferrin localization was correlated with the synthesis of a specific transcript, resulting from an alternative splicing, which leads to the elimination of the signal peptide sequence. These results suggest the existence of a functional difference between transferrin synthesized in the brain and in other organs such as liver and testis. They are in accordance with the hypothesis that transferrin plays a specific role, other than iron transport, in oligodendrocyte maturation and in the myelination process.

Nuclear receptors Nor1 and NGFI-B/Nur77 play similar, albeit distinct, roles in the hypothalamo-pituitary-adrenal axis.

Studies in Nur77-deficient mice have shown that the basal regulation of hypothalamic and pituitary functions as well as the adrenocortical steroidogenesis in these animals is normal. This indicates that Nur77-related orphan receptors may substitute Nur77 functions in the hypothalamo-pituitary-adrenal axis by a compensatory mechanism. Nor1 is the most recently cloned member of the NGFI-B/Nur77 subfamily, and its properties are still largely unknown. We demonstrate here that Nor1 is expressed in the pituitary gland and adrenal cortex, and that ACTH and angiotensin II (AngII) treatment of adrenal fasciculata cells induces Nor1 expression. Time-course analysis with both hormones on steroidogenic capacity and the specific gene expression in adrenal cells strongly suggest that Nor1 is an intermediate in the long-term consequences of ACTH or AngII treatment. The Nor1 and NGFI-B/Nur77 amino acid sequence homology and the analysis of the trans-activation properties of Nor1 show that the overall structural and functional organization of the two proteins is similar. As observed with NGFI-B/Nur77, Nor1 activates the expression of genes encoding steroidogenic enzymes as P450c21, through its interaction with NGFI-B response element promoter sequences. In contrast, binding experiments of Nor1 with the palindromic NurRE sequence suggest that Nor1 is not an efficient substitute for the NGFI-B/Nur77 activation of the POMC gene expression in pituitary glands. All these results indicate that Nor1 and NGFI-B/Nur77 may play similar albeit distinct roles in the hypothalamo-pituitary-adrenal axis. Further experiments also show that the mechanisms responsible for the transcriptional regulation of Nor1 in adrenal cells appear to depend on the protein kinase A and protein kinase C cascades.

Overexpression of the glucose-regulated stress gene GRP78 in malignant but not benign human breast lesions.

The 78 kDa glucose-regulated stress protein GRP78 is induced by physiological stress conditions such as hypoxia, low pH, and glucose deprivation which often exist in the microenvironments of solid tumors. Activation of this stress pathway occurs in response to several pro-apoptotic stimuli. In vitro studies have demonstrated a correlation between induced expression of GRP78 and resistance to apoptotic death induced by topoisomerase II-directed drugs. We were interested in characterizing this protein in human breast lesions for potential implications in chemotherapeutic intervention. Surgical specimens of human breast lesions and paired normal tissues from the same patients were flash frozen for these studies. Total RNA and/or protein were extracted from these tissues and used in northern and/or western blot analyses, respectively, to quantify the relative expression of GRP78. Northern blot analysis indicated that 0/5 benign breast lesions, 3/5 estrogen receptor positive (ER+) breast tumors, and 6/9 estrogen receptor negative (ER-) breast tumors exhibited overexpression of GRP78 mRNA compared to paired normal tissues, with fold overexpressions ranging from 1.8 to 20. Western blot analyses correlated with these findings since 0/5 benign breast lesions, 4/6 ER+ breast tumors, and 3/3 ER- breast tumors overexpressed GRP78 protein with fold overexpressions ranging from 1.8 to 19. Immunohistochemical analysis of these tissues demonstrated that the expression of GRP78 was heterogeneous among the cells comprising different normal and malignant glands, but confirmed the overexpression of GRP78 in most of the more aggressive ER- tumors. These results suggest that some breast tumors exhibit adverse microenvironment conditions that induce the overexpression of specific stress genes that may play a role in resistance to apoptosis and decreased chemotherapeutic efficacy.

Angiogenesis and cancer: from basic mechanisms to therapeutic applications.

An American Association for Cancer Research Special Conference on Angiogenesis and Cancer was held October 1115, 2000, in Traverse City, Michigan, U.S.A., to discuss the latest developments in angiogenesis research and antiangiogenic therapies. More than 300 scientists representing academics, industry and government attended this international meeting, which was organized by Drs. Judah Folkman (Children's Hospital/Harvard Medical School, Boston, Massachusetts) and David A. Cheresh (The Scripps Research Institute, La Jolla, California). In a series of seven sessions, 25 speakers presented the latest discoveries in the molecular biology and anatomy of tumor vasculature that unveiled novel targets for antiangiogenic therapies. More than 90 posters also helped bridge basic mechanisms of angiogenesis with new therapeutic strategies in the fight against cancer.

Anatomical basis for the lateral approach to the fourth ventricle.

This study focused on the posterior inferior cerebellar artery bifurcation and branching patterns in the fissures around the fourth ventricle. The vertebral arteries in 25 unfixed human cerebellum were cannulated and injected with polyester colored resin. The suboccipital surface of the cerebellum was exposed and the cisterna magna main landmarks localized. The average distance was 12.6 mm between the tonsillovermian notches and 21.8 mm between the inferior tips of the tonsils. The mean vertical distance between these horizontal planes was 14.5 mm. The posterior inferior cerebellar artery was found in the cerebellomedullary fissure in 42 of 50 cerebellar hemispheres, in seven cases the artery was absent and in one it was hypoplastic. The mean outer diameter was 1.8 mm and the average length was 27.9 mm. The posterior inferior cerebellar artery presented four bifurcation point patterns: superomedial, superolateral, inferomedial, and inferolateral. These patterns were characterized into subtypes based on the courses of the vermian and tonsillohemispheric branches. The perforating and choroidal branches originating in these segments were also studied. The mean number of perforating branches per hemisphere was 5.1. The range of the length was 2-10 mm and the range of the outer diameter was 0.1-0.3 mm. An average of 4.6 choroidal arteries originated from the tonsillomedullary and telovelotonsillar segments, a mean of 4 arose from both vermian and tonsillohemispheric branches. This information will facilitate surgical planning in approaching the fourth ventricle as well as the interpretation of cerebellar infarcts in the posterior inferior cerebellar artery area.

Development and validation of a short-term clinical model for assessing calculus inhibitory agents.

The purpose of this project was to develop and validate an efficient, short-term clinical model for assessing topically-applied anticalculus agents. In this model, calculus development occurred within 14 days on both labial and lingual surfaces of the mandibular anterior teeth. Because of documented long-term clinical efficacy, pyrophosphate dentifrices were used to investigate the validity of the short-term calculus model for evaluating anticalculus agents. This paper provides the results of the final 3 studies conducted during the development of this model. For each study, the design consisted of two 14-day phases, i.e., a control phase and a treatment phase, separated by a 7-day washout phase. At the start of each phase, a prophylaxis was performed on the mandibular anterior teeth to remove all plaque and calculus. At the end of each phase, supragingival calculus formation on the labial and lingual surfaces of these same teeth was measured using the VMI scoring method. Twice a day during the control phase, subjects applied a control dentifrice undiluted to the test teeth with a custom-fitted toothshield and brushed only the exposed teeth with the same dentifrice. For the treatment phase, subjects were randomly assigned to groups balanced on the basis of control-phase calculus scores and then delivered the dentifrices using the toothshield as in the first phase. After 14 days, calculus formation occurred in all groups. However, the pyrophosphate dentifrice groups had significantly less calculus (16-30%) than the control dentifrice group. These studies demonstrated that this methodology permitted rapid formation of dental calculus, and by substantiating with anticalculus systems documented to have activity in long-term human trials, it is concluded that this short-term clinical model is valid for assessing anticalculus agents.

[Effects of partial colonic resection in rats]. / Estudo dos efeitos das ressecções colônicas parciais em ratos.

UNLABELLED: In order to study the effects of partial colectomy, 30 rats Wistar were divided into three groups: GI and GII had the cecun and the ileo cecal valve resected and proximal colectomy was performed in GI and distal in GII. GIII had distal colectomy without cecum resection. The length of the remain colon was of 5 cm in all groups. Colostomy was performed in GI, GII and GIII. Parameters evaluated: body weight, fecal composition concerning to water, lipids and proteins on preoperative time and on the 10th, 20th, 30th, 40th, 50th and 60th days of postoperative time. All statistical tests were conducted at a 5% two-sided risk level. The evaluation was made by analysis of variance techniques. CONCLUSIONS: Concerning to body weight, there is no significant difference on the late postoperative time in any group or when comparing the three groups on this same period. On late postoperative time, GI and GII had a large amount of fecal water, lipids and proteins when compared to the preoperative time and to GIII. On the 60th day there's no significant difference on the quantity of fecal water when comparing the three groups and the same occurs on the 50th day when considering the quantity of fecal proteins.

Reverse transcription-polymerase chain reaction-based methodology to quantify differential gene expression directly from microdissected regions of frozen tissue sections.

Quantitative differences in the expression of oncogenes are a critical feature of the cancer process. Several methods are currently available for assessing differential gene expression, but none can be used to determine quantitative changes in gene expression from small numbers of cells. The ability to conduct this type of quantitative analysis would be useful in the study of definable, early stages of carcinogenesis when very few cells are involved. We therefore developed a highly sensitive, slide-based technique that incorporates the benefits of in situ polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR) to quantify differential c-myc gene expression from liver tissue sections having either low or high levels of proliferating hepatocytes. To eliminate the need for isolating and quantifying mRNA, cells of interest were microdissected from frozen histological sections and their RNA directly subjected to RT-PCR amplification. These reactions were conducted in the presence of an internal RNA standard that was specifically designed to normalize differential RT and PCR efficiencies between samples. GENESCAN software analysis was used to determine the ratios of the RT-PCR products of the target gene to the RNA standard. These ratios were then normalized to the numbers of cells isolated, as quantified by image analysis, and comparative gene expression values were determined between sample groups. We conclude that this technology can be adapted to study any gene of interest in any type of frozen tissue or isolated cells. This methodology is particularly applicable to the molecular analysis of histopathologically distinct preneoplastic and neoplastic lesions identified on tissue sections.

Malignant brain tumors in the elderly.

The increase in the incidence of primary brain tumors in the elderly over the last decade represents a serious problem because of the severe disability and risk of death associated with this disease. Surgery remains the mainstay of the management of malignant gliomas, and the magnitude of the resection correlates with the length of survival. The additional standard treatment modalities are reviewed. This article also discusses novel approaches being evaluated to treat both newly diagnosed and recurrent brain tumors.

Interactive image guidance in skull base surgery using an opto-electronic device.

The applicability of an image guidance frameless system based on an opto-electronic sensor device in skull base surgery was explored in this study. Five embalmed heads with external fiducial markers placed in noncoplanar points were scanned (CT scan) and different skull base approaches were reproduced in these specimens. The opto-electronic system is comprised of an infrared camera, a local rigid body, and a 24-light-emitting diode probe attached to different surgical instruments. DOS-based calibration and transformation software and Unix-based surgical planning software were also used. The anatomic landmarks identified during the dissection were matched with the corresponding points derived from computed tomographic (CT) scans. This information allowed the surgeon to develop a three-dimensional representation of the surgical field and to anticipate the next anatomic structure encountered during the dissection. This infrared device operated in real time, is not affected by external factors with regard to its accuracy, and does not interfere with standard neurosurgical techniques. This frameless system is helpful in minimizing the risk of morbidity and provides an accurate guide during the approach, as well as unobstructed access to the surgical field.

Permanent iodine-125 implants in the up-front treatment of malignant gliomas.

Between July 1989 and July 1992, 58 patients with newly diagnosed, histologically confirmed malignant gliomas (40 anaplastic astrocytomas, 18 glioblastoma multiforme) underwent implantation with low-activity iodine-125 sources. Patients were considered appropriate candidates for brachytherapy if their Karnofsky scores were > or = 70 and their contrast-enhancing tumors were < 6 cm in maximum diameter. Tumor volumes ranged from 0.1 to 90 ml. Ten patients had implants only. The other 48 patients received additional external beam radiation; 38 patients received radiation 1 to 2 weeks after the implant, and 10 patients received radiation preceding the implant. Median survival has not been reached but is currently greater than 31 months for patients with anaplastic astrocytoma and greater than 23 months for patients with glioblastoma. The rate of second operation for this group of patients was 45% (26 patients). Brain necrosis requiring resection occurred in 11 patients (19%). Although further follow-up is required, we conclude that low-activity permanent iodine-125 implants provide patients who have newly diagnosed malignant gliomas long-term survival with an acceptable risk of late complications.