Giant angiolipoma of the arm in an elderly patient.

Introduction Angiolipoma is a histological variant of lipoma and is the most common neoplasm in the trunk and extremities of young adults. It is extremely rare in elderly people, and its size is ≤4cm. Few data are available for large angiolipomas. Case History An 86-year-old patient was admitted to our surgical department due to a large mass on his left arm, which was resected. The specimen measured 19.5 × 15 × 10.5cm. Histopathological examination revealed a benign non-infiltrating angiolipoma. This is the first report of a giant angiolipoma of the arm reported in an octogenarian patient. Conclusions Giant lipomas of the upper extremities are extremely rare. Resection is associated with cure in most patients, but regular follow-up should be considered.

Preliminary crystallographic analysis of murine macrophage inflammatory protein 2.

Macrophage inflammatory protein 2 (MIP-2) has been crystallized by vapor diffusion of an 11 mg/ml protein solution in 100 mM-ammonium acetate against 30 to 40% polyethylene glycol (average molecular mass of 3350 Da). The crystals belong to space group P2(1)2(1)2(1) and have unit cell dimensions of a = 42.7 A, b = 59.3 A, and c = 100.3 A. The molecular mass of the protein and volume of the unit cell suggest that there are four monomers in the asymmetric unit. A data set to 2.3 A has been collected, and the self-rotation function identifies the presence of a non-crystallographic 2-fold axis.

Glucocorticoid counter regulation: macrophage migration inhibitory factor as a target for drug discovery.

Over the past year, human studies have confirmed and expanded the involvement of macrophage migration inhibitory factor (MIF) in a number of diseases that had originally been studied in animals. In addition to sepsis, rheumatoid arthritis, glomerulonephritis and inflammatory lung disease, elevated MIF levels have been described in patients suffering from ulcerative colitis, inflammatory neurological diseases and cancer. Cellular studies indicate that in addition to macrophages, MIF affects the activities of CD4+ and CD8+ T cells, natural killer cells, fibroblasts and endothelial cells, actions that may explain the contribution of MIF to inflammatory diseases and cancer. Molecular studies have identified direct interactions between MIF and several intracellular regulatory proteins (Jab1, PAG and p53) that control cellular growth and proliferation; however, how interactions with these proteins fit into a general scheme to explain MIF's biological activity has not been elucidated. The three-dimensional structure of MIF has offered some surprising clues and if the potential enzymatic sites identified are involved with MIF-associated diseases, they may provide good targets for therapeutic intervention.

Parastomal evisceration as an extremely rare complication of a common procedure.

The creation of an abdominal stoma is a common procedure performed as part of the treatment for many conditions. Common complications include poor stoma siting, high output, skin irritation, ischaemia, retraction, parastomal hernia and prolapse. An extremely rare stoma complication is parastomal evisceration. We present a case of a 48-year-old woman who presented to us with parastomal evisceration as a late complication of a transverse colostomy. It is the second case reported as a complication of this procedure but the first that occurred after such a long postoperative period (almost 18 months).

Functional and receptor binding characterization of recombinant murine macrophage inflammatory protein 2: sequence analysis and mutagenesis identify receptor binding epitopes.

Murine macrophage inflammatory protein-2 (MIP-2), a member of the alpha-chemokine family, is one of several proteins secreted by cells in response to lipopolysaccharide. Many of the alpha-chemokines, such as interleukin-8, gro-alpha/MGSA, and neutrophil activating peptide-2 (NAP-2), are associated with neutrophil activation and chemotaxis. We describe the expression, purification, and characterization of murine MIP-2 from Pichia pastoris. Circular dichroism spectroscopy reveals that MIP-2 exhibits a highly ordered secondary structure consistent with the alpha/beta structures of other chemokines. Recombinant MIP-2 is chemotactic for human and murine neutrophils and up-regulates cell surface expression of Mac-1. MIP-2 binds to human and murine neutrophils with dissociation constants of 6.4 nM and 2.9 nM, respectively. We further characterize the binding of MIP-2 to the human types A and B IL-8 receptors and the murine homologue of the IL-8 receptor. MIP-2 displays low-affinity binding to the type A IL-8 receptor (Kd > 120 nM) and high-affinity binding to the type B IL-8 receptor (Kd 5.7 nM) and the murine receptor (Kd 6.8 nM). The three-dimensional structure of IL-8 and sequence analysis of six chemokines (IL-8, gro-alpha, NAP-2, ENA-78, KC, and MIP-2) that display high-affinity binding to the IL-8 type B receptor are used to identify an extended N-terminal surface that interacts with this receptor. Two mutants of MIP-2 establish that this region is also involved in binding and activating the murine homologue of the IL-8 receptor. Differences in the sequence between IL-8 and related chemokines identify a unique hydrophobic/aromatic region surrounded by charged residues that is likely to impart specificity to IL-8 for binding to the type A receptor.

Accessibility of selenomethionine proteins by total chemical synthesis: structural studies of human herpesvirus-8 MIP-II.

The determination of high resolution three-dimensional structures by X-ray crystallography or nuclear magnetic resonance (NMR) is a time-consuming process. Here we describe an approach to circumvent the cloning and expression of a recombinant protein as well as screening for heavy atom derivatives. The selenomethionine-modified chemokine macrophage inflammatory protein-II (MIP-II) from human herpesvirus-8 has been produced by total chemical synthesis, crystallized, and characterized by NMR. The protein has a secondary structure typical of other chemokines and forms a monomer in solution. These results indicate that total chemical synthesis can be used to accelerate the determination of three-dimensional structures of new proteins identified in genome programs.

Salvaging recombinants from low-efficiency ligase reactions for more efficient subcloning.

Certain types of ligase reactions can be problematic, such as those involving PCR products, blunt-ends and multiple DNA inserts. A simple PCR-based strategy was developed to overcome cloning difficulties with these inefficient ligase reactions. After an initial ligase reaction, primers complementary to the vector are utilized to amplify the DNA fragment from (the few) successful recombinants in the ligation mixture. This DNA fragment is processed for use in a more conventional and straightforward ligase reaction. We demonstrate the potential of the technique by applying it to a variety of difficult ligase reactions.

Human papillomavirus testing and the outcome of treatment for cervical intraepithelial neoplasia.

OBJECTIVE: To investigate whether human papillomavirus (HPV) testing could be used in the follow-up after large loop excision of the transformation zone (LLETZ) for cervical intraepithelial neoplasia (CIN). METHODS: We performed a retrospective study of 41 women who developed subsequent CIN after LLETZ (group A) and 82 women without CIN for a minimum of 5 years after LLETZ (group B). The first post-treatment cervical smear was retrieved and examined for high-risk HPV deoxyribonucleic acid. The sensitivity, specificity, positive and negative likelihood ratio of HPV testing, first post-treatment Papanicolaou test, and excision margins for the detection of treatment failure were calculated. Multiple logistic regression analysis was also done. RESULTS: The HPV test was positive in 38 of 41 women in group A and 13 of 82 in group B (P <.001). An abnormal cytologic result in the first post-treatment smear was found in 20 of 41 in group A and 11 of 82 in group B (P <.001). Sixteen women in group A and 18 in group B had involved margins (P =.046). Values for the sensitivity, specificity, and positive and negative likelihood ratios of the HPV test were 93%, 84%, 5.8, 0.08; for the Papanicolaou test they were 49%, 87%, 3.9, 0.586; and for margin status they were 39%, 78%, 1.8, 0.782, respectively. Positive HPV test presents significantly high odds ratio for treatment failure (P <.001), independent of cytology and margin status. CONCLUSION: Women who postoperatively have positive HPV testing are at higher risk of treatment failure. This could be performed at the first post-treatment visit and further follow-up could be adjusted accordingly.

Cervical intraepithelial neoplasia outcomes after large loop excision with clear margins.

OBJECTIVE: To identify risk factors for residual or recurrent cervical intraepithelial neoplasia (CIN) after large loop excision of the transformation zone with clear margins. METHODS: We did a case-control study of women treated with loop excision for CIN who had adequate follow-up and in whom margins were believed to be clear. Women with clear margins in whom no subsequent lesions were found (controls) were compared with women who presented with subsequent CIN (cases). Epidemiologic and colposcopic risk factors for recurrence were analyzed. Multiple logistic regression analysis was done to identify independent risk factors. RESULTS: In 31 of 635 women studied (4.9%), subsequent lesions were diagnosed. Univariate analysis identified glandular involvement, satellite lesions, and age over 40 years in cases as significant. Multiple logistic regression analysis confirmed that these three characteristics were independent risk factors, with odds ratios of 4.9 (95% confidence intervals 1.9, 12.3), 19 (7.5, 48.2), and 6.7 (2.8, 15.8), respectively. Subsequent lesions were identified by colposcopy or cytologic testing during the first postoperative year in all but one case. CONCLUSION: Age over 40 years, glandular involvement, and satellite lesions were related to the reappearance of CIN after loop excision with clear margins. These findings could be used to define appropriate follow-up protocols.

Structural studies of chemokines that inhibit HIV-1 entry.

The chemokine receptors CCR5 and CXCR4 have emerged as essential mediators of HIV-1 pathophysiology, functioning as co-receptors for viral entry into cells. The physiological agonists of these receptors inhibit HIV-1 infection in vitro. The discovery of small molecules that disrupt the interactions between HIV-1 and chemokine receptors is one strategy to limit the spread of the virus. These compounds will complement already existing therapies that include HIV-1 reverse transcriptase and protease inhibitors. The complete structural elucidation of a chemokine ligand-receptor complex would be valuable for rational drug design, but has yet to be achieved. Structural studies of chemokine agonists and antagonists can also be useful in understanding interactions that may be important for drug optimization. This review examines the surface properties of the chemokine ligands human SDF-1alpha and HHV-8 vMIP-II, with a goal of determining receptor-interacting sites. In combination with site-directed mutagenesis of the chemokines and structure-activity relationships of chemokine-based peptides, this approach will lead to a better understanding of the interactions in the chemokine ligand-receptor system.

Transvaginal uterine ultrasonography compared with endometrial biopsy for the detection of endometrial disease in perimenopausal women with uterine bleeding.

BACKGROUND: Almost 70% of all gynecological consultations in perimenopausal women are related to irregular uterine bleeding. In this prospective study, we compared endometrial assessment by transvaginal ultrasonography (TVS) in perimenopausal women with irregular uterine bleeding to histological assessment and tested whether the TVS was effective as a diagnostic tool for the detection of endometrial pathology in these women. MATERIALS AND METHODS: Eighty consecutive perimenopausal women complaining of irregular uterine bleeding participated in the study. The women were evaluated by transvaginal scans, performed immediately before endometrial biopsy. The ultrasonographic results were compared with the histological diagnosis obtained from the endometrial biopsy. RESULTS: Sixty-seven out of 80 women (83.7%) had normal histological findings, whereas 13 (16.3%) had abnormal findings. No endometrial cancer was diagnosed in this cohort of women. In the 67 women with a histological diagnosis of normal endometrium, mean+/-SD endometrial thickness was 10.5+/-4.0 mm (range 4.0-18.5 mm), whereas the corresponding value in the 13 women with abnormal findings was 18.7+/-3.8 mm (range 13.5-22.5 mm). If a 13 mm cut-off limit was used for endometrial thickness, which would include all abnormal cases, the sensitivity, specificity and positive predictive values were 100%, 71.64% and 40.62%, respectively. CONCLUSION: TVS can identify women with perimenopausal bleeding in which the likelihood of endometrial pathology is high and in which tissue sampling should be performed. Thus, TVS can be a primary method of selecting women with perimenopausal bleeding who must be further investigated with more invasive methods such as endometrial biopsy.

Screening transvaginal uterine ultrasonography for identifying endometrial pathology in postmenopausal women.

BACKGROUND: During the last decade, transvaginal ultrasonography (TVS) has become a widely-used technique for the evaluation of endometrial histology. The purposes of this study were to compare transvaginal sonographic evaluation of the endometrium with histology obtained by endometrial biopsy in asymptomatic postmenopausal women and to determine whether screening transvaginal sonography might be useful in the evaluation of postmenopausal women. MATERIALS AND METHODS: The study included 59 unselected asymptomatic postmenopausal women who attended the outpatient clinic for annual cervical cytology at Ioannina University Hospital Greece. The women were evaluated by transvaginal scans, performed immediately before endometrial biopsy. RESULTS: In the 43 women with a histopathological diagnosis of normal endometrium/inadequate for assessment/atrophy, the mean endometrial thickness was 5.1 +/- 3.3 mm (range 0.8-13.8 mm) whereas the corresponding value in the 16 women with abnormal findings was 17.6 +/- 4.3 mm (range 9.4-24.6 mm) (p<0.001). If a 9 mm cut-off limit was used for endometrial thickness, the sensitivity, specificity and positive predictive value were 100%, 90.69% and 80%, respectively. CONCLUSION: TVS is a sensitive test for determining endometrial disease in asymptomatic postmenopausal women. However, well-designed studies should be conducted, completed, analysed and validated before a mass-screening program using TVS is implemented.

Screening transvaginal uterine ultrasonography for identifying endometrial pathology in postmenopausal women.

BACKGROUND: During the last decade, transvaginal ultrasonography (TVS) has become a widely-used technique for the evaluation of endometrial histology. The purposes of this study were to compare transvaginal sonographic evaluation of the endometrium with histology obtained by endometrial biopsy in asymptomatic postmenopausal women and to determine whether screening transvaginal sonography might be useful in the evaluation of postmenopausal women. MATERIALS AND METHODS: The study included 59 unselected asymptomatic postmenopausal women who attended the outpatients' clinic for annual cervical cytology at Ioannina University Hospital, Greece. The women were evaluated by transvaginal scans, performed immediately before endometrial biopsy. RESULTS: In the 43 women with a histopathological diagnosis of normal endometrium/inadequate for assessment/atrophy, the mean endometrial thickness was 5.1 +/- 3.3 mm (range 0.8-13.8 mm) whereas the corresponding value in the 16 women with abnormal findings was 17.6 +/- 4.3 mm (range 9.4-24.6 mm) (p<0.001). If a 9-mm cut-off limit was used for endometrial thickness, the sensitivity, specificity and positive predictive value were 100%, 90.69% and 80%, respectively. CONCLUSION: TVS is a sensitive test for determining endometrial disease in asymptomatic postmenopausal women. However, well-designed studies should be conducted, completed, analysed and validated before a mass-screening program using TVS is implemented.

Replacement of the uterus by malignant mesothelioma of the peritoneum: a case report.

Malignant mesothelioma of the peritoneum is a rare tumor that must be distinguished from the more common primary peritoneal serous epithelial neoplasms. We report a case of a 66-year-old female presenting with weight loss, anemia, and a large pelvic mass on ultrasound. At laparotomy a large (9 x 6.5 x 3.5 cm) mass in the anatomical position of the uterus was found in the minor pelvis. The uterus, tubes and ovaries could not be identified. Palpable paraaortic lymph nodes and liver nodules were found. Immunohistochemically the tumor cells were positive for cytokeratin, epithelial membrane antigen and vimentin-CEA, but S-100 protein and Leu-M1 were negative. Remnants of the uterine corpus, fallopian tubes and ovaries could not be identified. The overall features were best regarded as malignant peritoneal mesothelioma. There have been reports of mesotheliomas involving the uterus. However, this is the first reported case of mesothelioma causing total replacement of the uterus.

An observational study of women with positive HPV-DNA tests and normal cytology and colposcopy.

PURPOSE: High risk human papillomaviruses (HPV) are implicated in the aetiology of malignant cervical disease. The usefulness of HPV DNA tests in identifying women at risk of cervical cancer as an adjunct to cervical cytology is under evaluation. PATIENTS AND METHODS: This is a retrospective analysis of 47 women positive for high risk HPV but with negative cytology and negative colposcopy at the start of the study. Women were observed for three years or more (in 96% cases) using six-monthly combined HPV DNA tests, cytological and colposcopic evaluation. RESULTS: At the end of follow-up, 29/47 (62%) women were still positive for high risk HPV, 45/47 (96%) women had normal cytology and 47/47 (100%) women continued to have normal colposcopy. CONCLUSIONS: Normal colposcopy has an excellent negative predictive value for HPV positive women with normal cytology. These women can be safely screened cytologically on a three-yearly basis.

The use of transvaginal sonography and vaginoscopic hysteroscopy in women on tamoxifen.

BACKGROUND AND OBJECTIVES: Long-term administration of tamoxifen causes endometrial changes. The aim of this study was to evaluate the role of transvaginal sonography and vaginoscopic hysteroscopy in the screening of patients on tamoxifen. METHODS: Seventy patients with breast cancer treated with tamoxifen 20 mg daily underwent transvaginal sonography and vaginoscopic hysteroscopy, a modified relatively painless approach, at the beginning of the treatment and at a follow-up visit approximately 9 months after its initiation. RESULTS: At the follow-up visit, the mean uterine dimensions and mean endometrial thickness as measured by ultrasound were significantly larger, and pulsatility and resistance indices of the uterine arteries as measured by Doppler were significantly lower. Sonography revealed abnormal endometrial thickness in 73% (51 of 70) of the patients, and 83% (58 of 70) had hysteroscopical changes. Sonography missed 1 case of endometrial adenocarcinoma. CONCLUSIONS: Vaginoscopic hysteroscopy, an approach that causes reduced pain, can add significantly to the sensitivity of transvaginal sonography for the detection of endometrial changes in patients with breast cancer receiving tamoxifen. It is recommended for every patient prior to the initiation of treatment and at the follow-up visits.

[Abnormal ramification of the axillary artery: sub-scapular common trunk]. / Ramification anormale de l'artère axillaire: tronc commun sous-scapulaire.

In a man's dissected cadaver we observed an unusual branching of the right axillary artery, which gives a large collateral branch. This branch is the origin of several important arteries as the subscapular artery (with its usual branches), the anterior and posterior circumflex humeral arteries, the proDunda brachii a. and the ulnar collateral artery. The frequency of this variation is evaluated to be 0.45% and is comparable to those found in the literature. We propose to name this artery as common subscapular trunk. The trunk described by us possesses a diameter equal to 6.30 mm and is larger than the continuation or the axillary artery, which courses as superficial brachial artery.

Mechanisms of ketamine action on lipid metabolism in rats.

BACKGROUND AND OBJECTIVE: This study was conducted to determine the effect of ketamine on metabolic homoeostasis and particularly in lipoprotein lipase (LPL) activity in adipose tissue. METHODS: Sixty male Wistar rats were divided into six groups of 10 each. Group A served as controls, while Groups B-F received, respectively, ketamine 60, 80, 100, 120 and 140 mg kg(-1) intraperitoneally. The animals were sacrificed 20 min after the administration of ketamine. Insulin concentrations in plasma and total cholesterol, triglyceride, high-density lipoprotein (HDL) and free fatty acid (FFA) concentrations in serum were measured. LPL activity in adipose tissue and medium-chain acyl-CoA dehydrogenase (MCAD) content in muscle were determined. RESULTS: FFA concentrations in serum significantly increased from the second lowest dose of ketamine. Insulin concentrations in plasma did not exhibit any significant difference between groups. MCAD levels were 0.5-fold more in Group F than in Group A, while there were no significant differences between control group and Groups B-E. Furthermore, high concentrations (120 and 140 mg kg(-1)) of ketamine interfered with in metabolic homoeostasis by significantly reducing LPL activity, thus elevating triglyceride concentrations in serum without affecting cholesterol and HDL metabolism. CONCLUSIONS: Ketamine induces various metabolic effects due to changes in adipose LPL activity and MCAD levels in muscles. These findings seem to be significant only at high doses.

Crystal structure of macrophage migration inhibitory factor (MIF), a glucocorticoid-induced regulator of cytokine production, reveals a unique architecture.

The mediator originally described as a macrophage migration inhibitory factor (MIF) has been identified recently to be a novel anterior pituitary hormone and the first protein to be released from immune cells upon stimulation with glucocorticoids. Once released, MIF acts to override or counter-regulate the immunosuppressive effects of glucocorticoids on cytokine production in vitro and on LPS lethality in vivo. MIF also has been described to catalyze a tautomerization reaction, suggesting that an enzymatic activity may underlie certain of its biological properties. The three-dimensional crystal structure of human MIF has been solved at 2.6A resolution. The X-ray crystal structure reveals MIF to be a trimer of identical subunits. Each monomer contains two antiparallel alpha-helices that pack against a four-stranded beta-sheet. The monomer has an additional two beta-strands that interact with the beta-sheets of adjacent subunits to form the interface between monomers. The three beta-sheets are arranged to form a barrel containing a solvent-accessible channel that runs through the center of the protein along a molecular three-fold axis. Electrostatic potential maps reveal that the channel has a positive potential, suggesting that it binds negatively charged molecules. The elucidated structure for MIF is unique among cytokines or hormones and suggests that this glucocorticoid-induced counter-regulatory hormone has a novel mechanism of action.