bluuebunny/crossref_metadata_2025 huggingface dataset:
A Fast Amplification-Reverse Hybridization Assay Kit to Detect the Most Frequent Deficient Variants in the Alpha-1-Antitrypsin Gene
Authors: Zorzetto Michele , Tamburnotti Claudia , Maschietto Benedetto , Massi Guido , Battaggia Cinzia , Medaglia Sara , Luisetti Maurizio
Published: April 2003

Background: There is worldwide growing awareness of alpha-1-antitrypsin deficiency (AATD), a major hereditary disorder in Caucasians. The gold standard for its laboratory diagnosis is thin-layer isoelectric focusing, which should be performed in reference laboratories. Objectives: The aim of this study was to check the characteristics of a commercially available amplification-reverse hybridization assay kit in detecting at a molecular level the alpha-1-antitrypsin (AAT) Z and S variants, i.e. the most frequent variants associated with AATD, by comparing its performance with DNA restriction fragment length polymorphism. Methods: We studied samples from 36 subjects enrolled in the Italian National Registry for Severe Alpha-1-antitrypsin Deficiency. Based on previous plasma isoelectric focusing typing, we selected samples with the following phenotypes: MM (9 samples), MS (9 samples), SZ (3 samples), MZ (11 samples), ZZ (3 samples), and a rare variant (1 sample). DNA was extracted by the standard method. The presence of the AAT Z and S gene variants was determined by the amplification-reverse hybridization test kit, following the manufacturer’s instructions, and by the restriction fragment length polymorphism technique, according to established procedures. Results: We found that the identification of the AAT Z and S gene variants obtained by the amplification-reverse hybridization test kit was completely in agreement with that obtained by the restriction fragment length polymorphism technique. Conclusions: We conclude that the test kit provides a fast, easy and unambiguous identification of Z and S alleles. Because of its transferability to routine laboratories, the test kit may be useful in identifying cases of severe AATD, thus resulting in increasing awareness of this rare disorder.
The Influence of L-Arginine on the Regulation of Epidermal Arginase
Authors: Wohlrab Johannes , Siemes Christina , Marsch Wolfgang C.
Published: April 2003

Topical preparations containing urea are firmly established in dermatological therapy and nursing care therapy. In addition, urea is frequently used as an inactive ingredient with disinfecting, keratoplastic and penetration-promoting action in topical preparations. Despite good tolerance and ensured action, particularly the irritating effect on erosive, exudative or strongly inflammatory skin restricts its application. Endogenic urea is synthesised from L-arginine by an extra-hepatic arginase in keratinocytes. This enzymatic reaction is subject to regulation by manganese ions and the intracellular L-arginine concentration. L-Arginine is predominantly transported into the cell by a membrane transport system that is dependent on the membrane potential. By incubating keratinocyte cultures in different concentrations of L-arginine and manganese chloride, it could be shown that the keratinocytic urea synthesis can be increased. In relevant concentrations, L-arginine and manganese chloride do not exhibit any proliferation-inhibiting action, do not trigger any apoptosis or necrosis, and are stable. An increase in the expression of arginase cannot be detected using L-arginine. The application of L-arginine alone or in combination with manganese chloride increases the endogenous intrakeratinocytic urea synthesis and thus offers an option for topical therapeutic application in cases of dry skin conditions.
High-Frequency Ultrasound in the Evaluation of Pharmacological Effects on the Skin
Authors: Unholzer Angela , Korting Hans Christian
Published: April 2003

In 1987, the introduction of two 20-MHz B- (brightness-) scan ultrasound systems with axial and lateral resolution of 200 µm and 75–80 µm, respectively, rendered two-dimensional sonographic imaging in dermatology possible. Since then cutaneous sonography has increasingly been used to investigate drug effects on human skin. Both unwanted and wanted effects have been studied, including skin thinning due to repeated glucocorticoid application and reduction of skin thickening in hyperplastic disorders such as psoriasis vulgaris due to various treatment approaches. The resolution of 20-MHz ultrasound is sufficient for studying pathological changes and pharmacological effects in the dermis and the subcutaneous fat; for assessing the epidermis, frequencies between 50 and 100 MHz may prove to be more effective.
Natural Course of the Functional and Morphological Changes in Canine Autogenous Arterial Graft
Authors: Wei C.-G. , Komori K. , Mori E. , Kume M. , Yamaoka T. , Matsumoto T. , Sugimachi K.
Published: April 2003

Background: We examined the time course of endothelium-dependent and -independent responses in reversed autogenous arterial grafts during regeneration and tissue repair processes after arterial grafting in dogs. Materials and Methods: Autogenous arterial grafts implanted in the canine femoral artery were removed, cut into rings and suspended in organ chambers for isometric tension recording on the 1st, 3rd, 7th, 14th and 28th days after implantation. Electron-microscopic examination on the 1st, 3rd ,7th, 14th and 28th days after transplantation was also performed. Control arteries were taken from nonsurgically treated femoral arteries. Results: Acetylcholine (ACh) and A23187 caused endothelium-dependent relaxations in arterial grafts throughout the study. Although endothelium-dependent relaxations to ACh and A23187 on the 3rd day after transplantation were only significantly impaired compared to those of control, they were similar to the control within 1 week after grafting. Smooth muscle relaxations in response to sodium nitroprusside in arterial grafts throughout the study were comparable with those of control arteries. No apparent intimal thickening of the arterial grafts was observed. Electron microscopy scanning revealed mild endothelial cell damage in implanted autogenous arterial grafts on the 1st or 3rd day after grafting. Seven to 14 days after grafting, the endothelial cell layer appeared to be normal. Conclusions: It was observed that the endothelial function remained intact and there was an absence of intimal thickening in the arterial grafts. These observations may explain the improved patency of autogenous arterial grafts compared to the vein grafts in aortocoronary revascularization.
Evaluation of Subcutaneous Tissue Gases and pH during Induction of Acidosis and Alkalosis
Authors: Mellström Å. , Jedlinska Barbara , Hartmann M. , Jönsson K.
Published: April 2003

Peripheral tissue oxygen utilization was studied during hypoxic-induced acidosis and sodium bicarbonate-induced alkalosis in 8 domestic pigs by measurements of subcutaneous oxygen tension (PscO2), carbon dioxide tension (PscCO2) and pH (pHsc) in relation to central hemodynamic parameters and oxygenation. Hypoxic-induced acidosis resulted in a decrease in PscCO2 and arterial oxygen tension (PaO2) to one third of baseline values (p < 0.05), an increase in PscCO2 and arterial carbon dioxide tension (PaCO2) from 41 to 55 and 34 to 39 mm Hg, respectively (p < 0.05), and a decrease in pHsc from 7.47 to 7.30 (p < 0.05). PscO2 and PaO2 increased during reversal of hypoxia and infusion of bicarbonate (p < 0.05), without reaching baseline values. In parallel PscCO2 decreased and pHsc increased but changes lagged behind changes in blood gases. Alkalosis established by further infusion of bicarbonate resulted in a decrease in PaO2 to 62 mm Hg whereas PscO2 remained below baseline values (p < 0.05). Correction of oxygen utilization in the subcutaneous tissue as measured by the markers PscCO2 and pHsc is slower than indicated by changes in tissue oxygen tension, blood gases and pH. Overcompensation of acidosis with bicarbonate resulting in alkalosis impairs oxygenation.
Cytokine Profile of Serum and Bronchoalveolar Lavage Fluids following Thoracic Esophageal Cancer Surgery
Authors: Sato N. , Koeda K. , Kimura Y. , Ikeda K. , Ogawa M. , Saito K. , Endo S.
Published: April 2003

Objectives: The body’s response to stress is comprised of two opposing reactions, namely inflammation and immunosuppression. The balance between these two reactions not only changes over time, but also varies among different cells or organs. Limited information is available regarding the cytokine balance of circulating blood and inflammatory sites after thoracic surgical stress. Design: The subjects of the present study were 10 patients undergoing esophageal cancer surgery which requires thoracolaparotomic manipulation. The postoperative levels of interleukin (IL)-8, IL-10, IL-1 receptor antagonist (IL-1ra) and soluble tumor necrosis factor receptor I (sTNF-RI) in bronchoalveolar lavage fluids (BALFs) and peripheral blood were measured by enzyme-linked immunosorbent assay. The absolute concentration of cytokines in lower respiratory tract ([cytokine]LRT) was estimated by using the ratio of urea concentration in BALF and serum. Results: The levels of these cytokines in peripheral blood peaked within 24 h of start of the surgery (IL-8 85.6 ± 29.8 pg/ml; IL-10 65.1 ± 10.6 pg/ml; IL-1ra 2,807.8 ± 652.8 pg/ml, and sTNF-RI 3,996.3 ± 380.1 pg/ml). The level of [IL-8]LRT immediately after surgery was approximately 20 times higher than that in peripheral blood, and the level of [IL-1ra]LRT was approximately 4 times higher. In contrast, the level of [IL-10]LRT immediately after surgery was comparable to that in peripheral blood, but the level of [sTNF RI]LRT immediately after surgery was approximately one fifth of that in peripheral blood. Conclusions: The balance between pro- and anti-inflammatory reactions varies from one part of the body to the next, and changes over time after surgery. The profile of stress-induced pro-inflammatory cytokines and anti-inflammatory cytokines should be analyzed further to establish appropriate and effective cytokine modulatory therapeutic approaches.
A Method Using Laser Doppler Flowmetry to Study Intestinal and Pancreatic Perfusion during an Acute Intestinal Ischaemic Injury in Rats with Pancreatitis
Authors: Phillips A.R.J. , Farrant G.J. , Abu-Zidan F.M. , Cooper G.J.S. , Windsor J.A.
Published: April 2003

Intestinal ischaemia is implicated in the pathogenesis of severe acute pancreatitis, a disorder characterized by acinar necrosis. To study the relationship between pancreatic and intestinal microvascular perfusion during 40 min of intestinal ischaemia and 30 min of reperfusion in rodents with acute pancreatitis, a model utilizing laser Doppler flowmetry was developed. It is reported here together with practical solutions for (1) a modified method of vessel cannulation; (2) a novel method for the temperature-controlled optical coupling between laser Doppler probes and rodent tissues, and (3) a simple technique of inducing intestinal ischaemia-reperfusion while continuously monitoring the microvascular perfusion in both pancreas and intestine. The utility of the model is demonstrated in a pilot study that showed that the pancreatic perfusion fell acutely to 58% (p = 0.029) of baseline during the intestinal reperfusion phase. This reduced perfusion continued for 30 min despite recovery of both the intestinal perfusion and the mean arterial blood pressure to baseline levels.
Global Analysis of Gene Expression: Methods, Interpretation, and Pitfalls
Authors: Fryer Ryan M. , Randall Jeffrey , Yoshida Takumi , Hsiao Li-Li , Blumenstock Joshua , Jensen Katharine E. , Dimofte Tudor , Jensen Roderick V. , Gullans Steven R.
Published: April 2003

Over the past 15 years, global analysis of mRNA expression has emerged as a powerful strategy for biological discovery. Using the power of parallel processing, robotics, and computer-based informatics, a number of high-throughput methods have been devised. These include DNA microarrays, serial analysis of gene expression, quantitative RT-PCR, differential-display RT-PCR, and massively parallel signature sequencing. Each of these methods has inherent advantages and disadvantages, often related to expense, technical difficulty, specificity, and reliability. Further, the ability to generate large data sets of gene expression has led to new challenges in bioinformatics. Nonetheless, this technological revolution is transforming disease classification, gene discovery, and our understanding of regulatory gene networks.
Renal Gene Expression in Embryonic and Newborn Diabetic Mice
Authors: Wallner Elisabeth I. , Wada Jun , Lin Sun , Pan Xiaomin , Reddy Janaradan K. , Chugh Sumant S. , Kanwar Yashpal S.
Published: April 2003

Several novel genes that are upregulated in diabetic kidneys have been identified. Recently, transforming growth factor beta driven secreted proteins, i.e., connective tissue growth factor and gremlin (bone morphogenetic protein 2), have been identified, and their expression has been correlated with the tissue changes seen in diabetic nephropathy in the adult population. However, there are very few studies reported in the literature that describe the gene expression in the diabetic state during embryonic and neonatal life. It is well known that exposure to glucose or its epimer, i.e., mannose, induces marked dysmorphogenesis of the embryonic metanephros in an organ culture system. These changes are associated with ATP depletion and marked apoptosis, suggesting an oxidant stress in the induction of dysmorphogenesis of the embryonic metanephros. In view of the glucose-induced changes in the fetal metanephros, a diabetic state was induced by the administration of streptozotocin during pregnancy, and newborn mouse kidneys were processed for suppression subtractive hybridization-PCR. In addition, a diabetic state was induced in newborn diabetic mice, and after 1 week their kidneys were harvested and subjected to representational difference analysais of cDNA. Four novel genes with upregulated mRNA expression were identified. They included: (1) a translocase inner mitochondrial membrane 44 that is involved in the ATP-dependent import of preproteins from the cytosol into the mitochondrial matrix; (2) a kidney-specific aldo-keto reductase that utilizes NADPH and NADH as cofactors in the reduction of aromatic aldehydes and aldohexoses; (3) Rap1b, a Ras-related small GTP-binding protein that behaves as a GTPase and cycles between GTP-bound (active) and GDP-bound (inactive) states associated with conformational change, and (4) a fusion protein of ubiquitin polypeptide and ribosomal protein L40 (UbA52 or ubiquitin/60) that is intimately involved in the ubiquitin-dependent proteasome pathway related to the accelerated degradation of proteins under various stress conditions, such as those seen in patients with cancer and diabetes mellitus.
Field Kit to Characterize Physical, Chemical and Spatial Aspects of Potential Primate Foods
Authors: Lucas P.W. , Beta T. , Darvell B.W. , Dominy N.J. , Essackjee H.C. , Lee P.K.D. , Osorio D. , Ramsden L. , Yamashita N. , Yuen T.D.B.
Published: April 2003

An outline is given for a field kit aiming to substantially increase the in situ knowledge gleaned from feeding studies of primates. Measurements are made of colouration (spectrum of non-specular reflection) and many mechanical, chemical and spatial properties of primate foods.
Interspecific Variation of Scent-Marking Behaviour in Wild Tamarins, Saguinus mystax and Saguinus fuscicollis
Authors: Heymann Eckhard W.
Published: April 2003

The scent-marking behaviour of sympatric moustached, Saguinus mystax, and saddle-back tamarins, Saguinus fuscicollis, was compared in order to explore interspecific differences and potential sources of variation. The author examined basic patterns of scent marking (types, intensity, complexity), substrate use (type, orientation, height), and social patterning of scent marking in three groups of S. mystax and one group of S. fuscicollis at the Estación Biológica Quebrada Blanco, Peruvian Amazonia. S. mystax and S. fuscicollis differed significantly in the relative frequency of different types, and in the intensity and complexity of scent marking. Only S. fuscicollis showed allomarking. They also differed significantly in the type, orientation and height of substrates used for scent marking which corresponded to general differences in substrate use. In S. fuscicollis, but very rarely in S. mystax, two or more group members marked the same site sequentially or simultaneously. ‘Collective scent marking’, i.e. simultaneous scent marking by most or all group members, occurred only in S. fuscicollis. Since both tamarin species live sympatrically in mixed-species groups, ecological factors are unlikely to account for the differences found in scent-marking behaviour (except for differences in substrate use). They probably relate to as yet unknown differences in social and reproductive strategies of the two species.
Growth Patterns and Final Height in Congenital Adrenal Hyperplasia due to Classical 21-Hydroxylase Deficiency
Authors: Hargitai Gábor , Sólyom János , Battelino Tadej , Lebl Jan , Pribilincová Zuzanna , Hauspie Roland , Kovács József , Waldhauser Franz , Frisch Herwig
Published: April 2003

Background: Longitudinal growth and bone age (BA) development are the most important clinical parameters for monitoring adequate glucocorticoid replacement in children with congenital adrenal hyperplasia (CAH). Aim of the Study: To analyze the growth pattern of patients treated for CAH of the salt wasting (SW) and simple virilizing (SV) clinical forms; to evaluate final height as compared to reference data and individual target height; to evaluate the course of BA development. Patients and Methods: A large database of 598 patients with CAH was created in 5 Central European countries and growth data of 341 treated patients with 21-hydroxylase deficiency were analyzed retrospectively. The patients were of Caucasian origin. Centiles were constructed in a cross-sectional manner and an additional longitudinal analysis was performed in order to evaluate the pubertal growth spurt by applying particular statistical methods (Preece-Baines model). Results: The growth of SW CAH patients was impaired in infancy and early childhood (0–3 years of age), but followed normal patterns in childhood until puberty. In contrast, children with SV CAH had normal patterns of growth in infancy and early childhood and were considerably taller than healthy references during childhood. In the longitudinal study, peak height velocity in both boys and girls was normal, but it occurred at an earlier age than in the standard population. The final height of patients with CAH was reduced in comparison to both the reference and the individual target height. No correlations were found between final height and age at the start of the therapy in SV patients or between final height and year of birth. BA was advanced in both types of CAH, but more accelerated in SV patients. Conclusion: Characteristic growth patterns for treated SV and SW CAH children were identified, with a normal pubertal growth spurt and reduced final height being observed.
Effect of Low Birth Weight on Adrenal Steroids and Carbohydrate Metabolism in Early Adulthood
Authors: Szathmári Miklós , Vásárhelyi Barna , Tulassay Tivadar
Published: April 2003

Aim: Data are inconsistent whether hyperinsulinemia might be associated with adrenal hyperandrogenism in young adults born with low birth weight (LBW). Method: We investigated the insulin and adrenal steroid production of 70 young LBW adults [33 women (birth weight: 1,795 ± 435 g) and 37 men (birth weight: 1,832 ± 337 g)]. Their results were compared to those of 30 controls (14 men, 16 women), born with normal weight. Results: In LBW women, we measured higher basal DHEA (33.5 ± 13.1 vs. 23.6 ± 8.7 nmol/l, p < 0.05), DHEAS (8.0 ± 2.3 vs. 6.3 ± 2.1 µmol/l, p < 0.05), androstenedione (8.3 ± 2.8 vs. 6.0 ± 2.2 nmol/l, p < 0.05) and cortisol (0.25 ± 0.07 vs. 0.20 ± 0.07 µmol/l, p < 0.05) levels and higher insulin response during oral glucose tolerance test (log.AUCins: 2.62 ± 0.06 vs. 2.57 ± 0.03, p < 0.05). DHEA levels correlated with fasting insulin levels (r = 0.45, p < 0.01) and insulin response (r = 0.33, p < 0.05). In LBW men, higher cortisol (0.27 ± 0.06 vs. 0.22 ± 0.06 µmol/l, p < 0.01) and SHBG (18.4 ± 10.4 vs. 12.7 ± 5.9 nmol/l, p < 0.05) levels were found. Conclusions: Our results suggest that modest hypercortisolism is present in young LBW adults. While the endocrine sequel of hypercortisolism raised insulin response and hyperandrogenism is detectable in apparently healthy young LBW women, it is absent in young LBW men. This suggests that gender-dependent mechanisms might play a role in the development of insulin resistance in LBW adults.
Molecular Analysis of CYP-21 Mutations for Congenital Adrenal Hyperplasia in Singapore
Authors: Loke Kah Yin , Lee Yung Seng , Lee Warren Wei Rhen , Poh Larry Kok Seng
Published: April 2003

Background: Congenital adrenal hyperplasia arising from 21-hydroxylase deficiency is associated with mutations in the CYP21 gene on chromosome 6p. This is the first report on the mutational spectrum of the CYP21 gene in Singapore. Methods: To catalogue the mutations, ten exons of the CYP21 gene from 28 Singaporean patients were analyzed by PCR amplification and direct sequencing. Results: Common mutations in descending order were the intron 2 splice site mutation (32.7% of the alleles), the I172N mutation (23.1% of the alleles), and the R356W mutation (19.2% of the alleles). Two potentially novel mutations were discovered: (1) duplication of 111 bp from codon 21 to codon 57 (exon 1) and (2) missense mutation (L261P, exon 7). There was generally a good genotype-phenotype correlation, allowing accurate prediction of the disease severity.
The Effect of Growth Hormone Treatment on Biochemical Indices in Hypophosphatemic Rickets
Authors: Darendeliler Feyza , Baş Firdevs , Karaaslan Neşe , Hekim Nezih , Bundak Rüveyde , Saka Nurçin , Günöz Hülya
Published: April 2003

Background: Growth hormone (GH) in combination with conventional therapy in hypophosphatemic rickets (HR) has been shown to promote renal phosphate (P) conservation and to result in a better metabolic control. This study aimed at investigating the acute biochemical effects of GH in 7 patients (5 female, 2 male) with HR aged between 2.16 and 16 years. Methods: Each patient received the following in a sequential design: oral P plus 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] therapy to determine the optimum doses for baseline requirement followed by GH-only therapy and GH +1,25-(OH)2D3 therapy and GH + P +1,25-(OH)2D3 therapy each for 2 weeks with 1 washout week off treatment in between. GH was given at a dose of 0.03 mg/kg/day s.c. on a daily basis. The dose of oral P used ranged between 500 and 2,000 mg/day, and the dose of 1,25-(OH)2D3 ranged between 0.25 and 0.5 µg/day and was kept constant for each child throughout the study. Results: Laboratory investigations repeated at the end of each treatment, and the first washout period showed that the serum P level was highest (2.9 ng/ml) during the GH + P + 1,25-(OH)2D3 period with higher serum 1,25-(OH)2D3 levels: 50.9 ± (SD) 23.4 ng/l. Parathyroid hormone and alkaline phosphatase levels did not show a significant difference between the periods. The tubular P reabsorbtion rate showed an insignificant increase during GH therapy periods. Conclusion: Considering the fixed dose of P and calcitriol, it may be concluded that GH added to conventional treatment in HR resulted in a slight improvement in the biochemical parameters without any side effects at the short term.
Recombinant versus Urinary Follicle-Stimulating Hormone in the Low-Dose Regimen in Anovulatory Patients with Polycystic Ovary Syndrome: A Safer and More Effective Treatment
Authors: Fulghesu Anna Maria , Apa Rosanna , Belosi Chiara , Ciampelli Mario , Selvaggi Jr. Luigi , Cucinelli Francesco , Caruso Alessandro , Mancuso Salvatore , Lanzone Antonio
Published: April 2003

Background: We studied polycystic ovarian syndrome (PCOS) in fifty 25- to 37-year-old women who failed to conceive with clomiphene citrate treatment. Methods: Twenty patients were submitted to treatment with low-dose (75 IU) urinary FSH (uFSH) in order to achieve ovulation and 30 patients were treated with recombinant FSH (rFSH) according to the same protocol. Results: Ovulation was achieved in 75 and 97% of the cycles after uFSH and rFSH, respectively (p < 0.02). The length of treatment needed to achieve ovulation, the number of ampules given and dose per kilogram were significantly lower in the rFSH group. Mild ovarian hyperstimulation syndrome (OHSS) was observed in 9 uFSH cycles, whereas only 1 of the women treated with rFSH developed an OHSS (1/38 vs. 9/36; p < 0.01). Conclusion: rFSH is more efficient than uFSH in inducing ovulation in PCOS patients. The high prevalence of ovulatory cycles using a lower dose guaranteed greater safety of treatment and significantly reduced the incidence of OHSS.
Uptake of pp65 in in vitro Generated pp65-Positive Polymorphonuclear Cells Mediated by Phagocytosis and Cell Fusion?
Authors: Kas-Deelen A.M. , The T.H. , Blom N. , van der Strate B.W.A. , De Maar E.F. , Smit J. , van Son W.J. , Harmsen M.C.
Published: April 2003

Objective: The cytomegalovirus (CMV) antigenemia consists of the detection of CMV pp65 in the nucleus of polymorphonuclear granulocytes (PMN), but it is unclear where and how PMN pick up virus particles or proteins. In an in vitro model for CMV antigenemia we investigated the mechanism of pp65 uptake by PMN that results in its expression in the nucleus. Methods: A series of inhibitors of different mechanisms was used to study the uptake of pp65 by PMN during coculture with CMV-infected endothelial cells and we performed a morphological analysis by light and transmission electron microscopy. Results: Nocodazole and cytochalasin B inhibited uptake of pp65 by PMN with 59.4 ± 14.1 and 73.3 ± 12.7%, respectively. The presence of anti-CMV hyperimmune globulin or lactoferrin during coculture reduced the number of pp65-positive PMN with 45.8 ± 7.0 or 40.6 ± 3.2%. Furthermore, a small number of the pp65-positive PMN obtained after coculture had fused to large cells with multilobed nuclei. PMN were observed that enclosed viral particles as well as free viral particles containing PMN in the cytoplasm. Conclusion: Fusion of viral particles with PMN and phagocytosis are both involved in the uptake of pp65.
BHRF1 Is Highly Conserved in Primate Virus Analogues of Epstein-Barr Virus
Authors: Williams Tracey , Sale Darleen , Hazlewood Sheila Ann
Published: April 2003

The Epstein-Barr virus (EBV) protein BHRF1 (BamHI rightward reading frame 1) was the first viral member of the Bcl-2 family of apoptosis-regulating proteins described. In vitro studies imply that BHRF1 is dispensable for virus-induced cellular transformation and virus replication. However, in contrast to several essential viral genes that show divergence outwith their functional domains, sequence data from a wide range of EBV isolates show there is striking conservation of the BHRF1 gene. Contrary to the in vitro studies, the high degree of conservation hints at a more important role for BHRF1. Analogous viruses are endemic in each of the higher primate species. Whilst their genome organisation is colinear, limited sequence analysis indicates that the viruses have diverged significantly and that only important functional domains of proteins are likely to be conserved. We have isolated the BHRF1 equivalents from the viruses which infect chimpanzees (Herpesvirus pan) and baboons (Herpesvirus papio) and find that they are highly homologous in both species, strengthening the hypothesis that BHRF1 plays a significant, evolutionarily conserved function in vivo and that changes to the protein are not well tolerated.
Recent Advances in the Development of Peptide Vaccines for Hepatitis B
Authors: Moynihan Jennifer S. , Howard Colin R.
Published: April 2003

The control of hepatitis B by vaccination is arguably one of medicine’s greatest achievements in terms of protecting infants and adults at high risk of infection. Paradoxically, however, the existence of a large reservoir of chronically infected individuals will not diminish the risk of infection by those coming into close contact with such persons until universal infant immunisation is practised globally and vaccines are in place to ensure maximum efficacy in those with impaired immune responses, immunity is achieved with fewer doses, and immunisation as an adjunct to the antiviral treatment of chronic carriers is adopted. These imperatives have continued to stimulate research into vaccines based on chemically synthesised short peptides, and those systems best suited for their delivery. This review discusses the potential of synthetic peptide formulations as efficient inducers of both humoral and cellular immune responses against hepatitis B, and reviews recent advances in peptide delivery. Synthetic peptide and delivery systems technologies will, amongst others, be of paramount importance in the global fight for the eradication of hepatitis B in the 21st century.
Murine Cytomegalovirus Infection Induces Cellular Folylpolyglutamate Synthetase Activity in Quiescent Cells
Authors: Cavallo Rossana , Lembo David , Gribaudo Giorgio , Landolfo Santo
Published: April 2003

Cytomegalovirus (CMV) infection stimulates the expression of cellular enzymes involved in the biosynthesis of DNA precursors. Among them, dihydrofolate reductase (DHFR) and thymidylate synthase (TS) require folate as coenzymes. In growing cells, folates are readily converted to polyglutamated forms by the cellular enzyme folylpolyglutamate synthetase (FPGS). Polyglutamated folates are selectively retained within the cell and have an increased affinity for DHFR and TS. Here we report that murine CMV (MCMV) increases the levels of the FPGS mRNAs as well as the enzymatically active FPGS protein through a mechanism that requires viral gene expression. FPGS induction by MCMV would provide the necessary supply of polyglutamated folates to the cellular enzymes involved in the biosynthesis of deoxyribonucleotides, enabling viral DNA replication to take place in quiescent cells.
Replication-Competent Herpes simplex Virus Type 1 Mutant Expressing an Autofluorescent Glycoprotein H Fusion Protein
Authors: Lorentzen E.U. , Eing B.R. , Hafezi W. , Manservigi R. , Kühn J.E.
Published: April 2003

The heterocomplex of glycoproteins H (gH) and L (gL) of herpes simplex virus type 1 (HSV-1) is essential for viral infectivity and is involved in viral penetration, cell-to-cell spread, and syncytium formation. We constructed an HSV-1 mutant expressing a gH-EGFP (enhanced green fluorescent protein) fusion protein under the control of the gH true late promoter. The EGFP coding sequence was cloned after the gH signal peptide into the HSV-1 genome. Superinfection of transfected, gH-nontranscomplementing cells with gH-negative HSV-1 resulted in a replication-competent recombinant virus. Cells infected with the recombinant virus exhibited strong and stable EGFP-specific fluorescence late in infection, and autofluorescence was detected in purified virions. The recombinant genotype of the mutant was confirmed by PCR. The 140-kD gH-EGFP fusion protein showed an N-linked glycosylation pattern similar to gH-1, was recognized by the conformation-dependent gH-specific monoclonal antibodies 52S and LP11 and formed a heterocomplex with gL which was transported to the cell surface and integrated into the viral envelope. Infectivity of the gH-EGFP mutant was neutralized by antibodies 52S and LP11. To our knowledge, this is the first replication-competent HSV-1 mutant expressing an autofluorescent essential glycoprotein which will be a versatile tool for studies of penetration, late gene expression, transport and tissue spread.
Identification of New Endogenous Retroviral Sequences Belonging to the HERV-W Family in Human Cancer Cells
Authors: Yi Joo-Mi , Lee Won-Ho , Kim Hwan-Mook , Kim Heui-Soo
Published: April 2003

A human endogenous retroviral family (HERV-W) has recently been identified on chromosome 7 that contains a single complete open reading frame putatively encoding an envelope protein. We have identified fifteen HERV-W families on human genomic DNA using a monochromosomal panel in our previous study. In order to identify additional families, we examined genomic DNA derived from cancer cell lines (A549, AZ521, OVCAR3, RT4) using the PCR approach. Five env fragments of a HERV-W family were newly identified and analyzed. They showed a high degree of nucleotide sequence similarity (93–97%) to that of the other HERV-W families. Translation of the env fragments showed no frameshift and termination codon by deletion/insertion or point mutation in clones A549-2, AZ521-3, and OVCAR-3. The ratio of synonymous to nonsynonymous substitutions indicated that negative selective pressure is acting on A549-2, AZ521-3, and OVCAR-3 sequences. These env gene sequences could be associated with an active provirus in human cancer cells (A549, AZ521, OVCAR3).
Characterization of DNA Binding Protein of Porcine Adenovirus Type 3
Authors: Zhou Yan , Pyne Caron , Tikoo Suresh K.
Published: April 2003

To identify and characterize the protein encoded by the E2A region of porcine adenovirus (PAV)-3, DNA sequence coding for a portion (amino acids 102–457) of the DNA binding protein (DBP) open reaching frame was cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase protein of Schistosoma japonica. The affinity-purified fusion protein was used to immunize rabbits. Immunoprecipitation/Western blot analysis demonstrated that the antisera specifically recognized a protein of 50 kD in PAV-3-infected cells. Immunoperoxidase staining detected the DBP protein predominantly in the nucleus of the cells. Western blot analysis demonstrated that DBP was detected as early as 6 h after infection and remained detectable throughout the infection. Based on these results, a novel assay for quantitation of PAV-3 was established. The assay is less time consuming and can be performed in different porcine cells. In addition, virus titers determined by this assay are comparable to the standard plaque assay.
Mapping of Epitopes on the Fiber Knobs of Human Adenovirus Serotypes 8 and 15
Authors: Liebermann Herbert , Lotz Kristina , Seidel Werner
Published: April 2003

In order to obtain information on the linear antigenic epitopes on fiber knobs of adenovirus serotypes 8 (Ad8) and 15 (Ad15) of subgenus D, the binding of polyclonal virus- and fiber-specific rabbit antibodies to overlapping peptides covering the fiber knob was studied. The main antigenic epitopes of the fiber knob of Ad8 (FK8) were represented by the peptides P4 [amino acids (aa) 213–227], P6 (aa 233–247), P11 (aa 283–297), P13 (aa 303–317) and P15 (aa 316–325); the peptides P1 (aa 183–197), P8 (aa 253–267), P10 (aa 273–287) and P23 (aa 340–349) were moderately reactive. The peptides P4, P6, P11, P13 and P15 span the β strands C, D, G and H, parts of the CD and DG loops and the complete GH loop of the fiber knob. The main epitopes of the fiber knob of Ad15 (FK15) were represented by peptides P5 (aa 198–212), P10 (aa 223–237), P12 (aa 233–247), P13 (aa 238–252), P26 (aa 303–317), P29 (aa 318–332) and P32 (aa 333–347), spanning the β strands B, D, G, H and I, partly strand C, the CD loop, parts of the AB and DG loops, the GH and HI loops and the N-terminal part of the IJ loop. The amino acid sequence alignment showed that the location of the linear FK8 and FK15 epitopes was found to be overlapping to a major extent. Two serotype-specific epitopes were determined on FK15, represented by P10 and P13.
Clinicopathologic Profiles and Prognosis of Gastric Carcinomas from the Cardia, Fundus/Body and Antrum
Authors: Pinto-de-Sousa J. , David Leonor , Seixas Mário , Pimenta Amadeu
Published: April 2003

Background/Aims: The putative influence of tumor location on the biologic behavior of gastric carcinomas remains controversial. The aim of this study was to investigate if carcinomas arising in the three types of gastric mucosa (cardia, fundus/body and antrum) have different clinical and pathologic profiles and carry a different prognosis. Methods: Three hundred and two patients with cardia or gastric carcinoma resected between 1984 and 1996 were retrospectively studied. Cases were divided in three groups according to tumor location: cardia (n = 80); fundus/body (n = 60); antrum (n = 162). The three groups were crosstabulated with clinic and pathologic parameters, such as age, sex, macroscopy, histology, desmoplasia, tumor size, depth of tumor wall penetration, nodal status, venous invasion and stage. Survival rates were calculated for the three locations according to the aforementioned parameters. Univariate survival analysis and Cox regression were performed for each location. Results: Cases from the cardia and fundus/body were similar and distinct from antrum cases according to macroscopy, tumor size, depth of wall penetration, venous invasion, nodal status and stage. Cases from fundus/body were similar to antrum cases and distinct from cardia cases according to gender and Laurén’s classification. An overall difference in survival between the three locations was observed (p = 0.006). Cumulative survival was better for patients with carcinomas in the antrum than in the cardia (p = 0.04) and in the fundus/body (p = 0.003); no significant differences were observed in survival between cardia and fundus/body carcinoma cases. Cox regression identified stage and venous invasion as prognostic factors for patients with carcinomas in the three locations. In the group of cardia tumors, older patients had a worse outcome and in the group of fundus/body carcinomas, large tumors were associated with a poorer survival. Conclusions: Our results show that cardia carcinoma and antrum carcinoma are distinct gastric carcinoma entities whereas fundus/body carcinoma shares some characteristics from both entities.
Three Cases of Small Bowel Intussusception in Relation to a Rare Lesion: Inflammatory Fibrous Polyps
Authors: Zager Jonathan S. , Shaw Jason P. , Kaufman Jarrod P. , DeNoto George
Published: April 2003

Background/Aims: Inflammatory fibrous polyps (IFPs), also known as inflammatory pseudotumors, occur rarely in the gastrointestinal tract. IFPs have variable presentations, often presenting as small bowel obstruction due to intussusception or, less commonly, as an incidental finding on radiological examinations or screening colonoscopies. The diagnosis and management of IFPs will be discussed through a review of the literature and a series of cases from our own institution. Methods: A retrospective analysis of the diagnosis, management and complications of IFPs was performed by a literature review. This was accompanied by a series of 3 cases of IFPs, 2 of which causing intussusception, diagnosed and treated in our own institution. Conclusions: IFP is a rare disease and has a variable presentation, from asymptomatic to small bowel obstruction due to intussusception. IFPs cannot be differentiated from malignancy without histological examination. Therefore, whether diagnosed incidentally or in the setting of intussusception, the treatment of IFPs is surgical resection of the involved bowel.
Preoperative Adjuvant Radiochemotherapy May Increase the Risk in Patients Undergoing Thoracoabdominal Esophageal Resections
Authors: Liedman Bengt , Johnsson Erik , Merke Claes , Ruth Magnus , Lundell Lars
Published: April 2003

Background/Aim: Fatigue and malaise are common symptoms after radiochemotherapy which could affect patients’ working capacity and add to the subsequent postoperative risk. The aim of the study was to investigate whether neoadjuvant radiochemotherapy in patients scheduled for a thoracoabdominal resection impairs their working capacity and adds to the postoperative risk. Patients and Methods: 29 patients with resectable tumors (T3 or N1) and a working capacity of ≧80 W on a bicycle test were included into the study. The tumor stage was determined by endoscopic ultrasound and computed tomography scan. The patients were given neoadjuvant radiochemotherapy during 38 days, consisting of two cycles of 5-fluorouracil (1,000 mg/m2) daily during 5 days and cisplatinum (100 mg/m2) on two occasions. Accelerated radiochemotherapy to a total dose of 40.8 Gy was given. Surgery was planned 4–6 weeks after completion of therapy. The patients had a bicycle test before induction of neoadjuvant treatment and 2–5 days prior to the operation. Results: Three patients died already during the course of neoadjuvant treatment. The 26 remaining patients who had a bicycle test decreased their working capacity by a mean of 30 W (p < 0.0001). Ten patients had a decrease of their working capacity to ≤90 W, of whom 6 died within 3 months postoperatively, and of the patients who had the working capacity decreased to ≤80 W, 4 out of 5 died during the corresponding postoperative period. None of the patients, who performed >100 W at the second preoperative bicycle test died. Conclusions: Preoperative adjuvant radiochemotherapy, according to the present schedule, clearly exerts a detrimental effect on the patients’ working capacity. A physical performance, at the time of the operation, ≤80–90 W strongly predicts the subsequent postoperative risk. Similar adjuvant therapies may, therefore, in fact be harmful to some patients scheduled for a thoracoabdominal resection.
Video Mediastinoscopic Division of a Congenital Esophagobronchial Fistula with Esophageal Diverticulum by a Paraesophageal Approach
Authors: Kido Tetsuo , Nakahara Masaaki , Imabun Shigeru , Nakao Kazuyasu
Published: April 2003

We treated a patient with an esophagobronchial fistula accompanied by stomach cancer. Using an endo-stapling technique, the fistula was divided video mediastinoscopically by a paraesophageal approach through the esophageal hiatus in the diaphragm.
Carcinosarcoma of the Esophagus – Pattern of Recurrence
Authors: Ziauddin Mohammed F. , Rodriguez Heron E. , Quiros Emily D. , Connolly Mark M. , Podbielski Francis J.
Published: April 2003

Carcinosarcoma of the esophagus is a rare malignant neoplasm, predominantly affecting men in their seventh decade of life. While presenting symptoms and anatomic location of squamous cell and carcinosarcoma of the esophagus are similar, the latter often presents as a large intraluminal polypoid mass on barium esophagram. The more favorable prognosis associated with carcinosarcoma versus other esophageal neoplasms has been attributed to early onset of symptoms, resulting in prompt diagnosis, and a lower propensity for tumor invasion. We report the case of an elderly woman presenting with dysphagia who was initially diagnosed with esophageal leiyomyosarcoma. Final tumor pathology showed esophageal carcinosarcoma.
Successful Treatment of Dissemination of Hepatocellular Carcinoma to the Pleura and Diaphragm after Percutaneous Liver Biopsy
Authors: Uenishi Takahiro , Kubo Shoji , Hirohashi Kazuhiro , Tanaka Hiromu , Ohba Kazuki , Kinoshita Hiroaki
Published: April 2003

Background/Aim: Treatment for dissemination of hepatocellular carcinoma to the pleura and diaphragm following percutaneous needle biopsy has not been established. Methods: The case of a 57-year-old man who underwent percutaneous needle biopsy for liver tumor is presented. Results: Ten months after resection of the tumor (moderately differentiated hepatocellular carcinoma), masses in the right pleural cavity and on the diaphragm were detected by computed tomography. Resections of the masses with surrounding tissue and the diaphragm and wedge resection of the right lung were performed. A wide range of the pleura and the diaphragm was coagulated with an argon beam coagulator. The patient is in good health without recurrence 4 years after the operation. Conclusion: Aggressive surgical treatment should be considered for patients with dissemination of hepatocellular carcinoma by needle biopsy when the lesions are limited.
Effects of Propranolol on Human Postoperative Ileus
Authors: Bandeira Ferraz Álvaro Antônio , Wanderley Geraldo José Paraíso , Arcanjo dos Santos Jr Miguel , Mathias Carlos Augusto , Corrêa de Araújo Jr José Guido , Machado Ferraz Edmundo
Published: April 2003

Background: The sympathetic nervous hyperactivity present in response to surgical stress has been implicated as an important component of the postoperative paralytic ileus. A randomized and prospective study was conducted, evaluating the effects of the preoperative beta-adrenergic blockade with propranolol in schistosomotic patients during the period of postoperative ileus. Methods: The study compared schistosomotic patients submitted, or not, to beta-adrenergic blockade. Basal cardiac frequency was determined and propranolol was used in a dose of 40 mg twice a day. The dose was adjusted weekly until a minimum decrease of 20% in cardiac frequency was achieved. Three coupled bipolar electrodes were placed in the left colon in both groups, and registration of myoelectric activity of the left colon was made twice a day during the period of postoperative ileus using a system of data collection (DATA Q Series 200). The electric signals were previously amplified, filtered and separated into Electric Control Activity (ECA) and Electric Response Activity (ERA). Results: The dose of propranolol varied from 80 to 160 mg/day. The proportional decrease in basal heart frequency varied from 20 to 33%, with an average of 25.4 ± 3.9% in the propranolol group, maintaining a mean of 24.3 ± 3.6% decrease in the postoperative period. Differences on clinical recovery of the postoperative ileus were not found. Significant differences on electromyographic patterns were not observed between the groups, except for the presence of a greater number of short-duration contractions in the second postoperative day in the beta-blocked group. Conclusion: The authors suggest that the preoperative beta-adrenergic blockade with propranolol does not determine myoelectric activity changes that could contribute to an earlier resolution of postoperative ileus.
Biliary Web – A Rare Cause of Extrahepatic Biliary Obstruction
Authors: Margolis Marc , Schein Moshe
Published: April 2003

A case of an isolated mucosal web of the common hepatic duct is presented. Such extrahepatic biliary webs are extremely rare causes of biliary obstruction, likely congenital in nature, but presenting in later life due to the initial patency of these webs in allowing bile drainage from the liver. The relevant literature is reviewed; diagnostic and therapeutic aspects are discussed.
Neuromuscular and Vascular Hamartoma of the Small Intestine
Authors: Scintu Francesco , Giordano Monica , Mascia Roberto , Comella Donatella , Casula Giuseppe
Published: April 2003

Neuromuscular and vascular hamartoma is an extremely rare stricturing condition of the small bowel. It consists of abnormal mixtures of intestinal tissues: disorganized fascicles of smooth muscle derived from the submucosa, bundles of nonmyelinated nerve fibers with scattered abnormal ganglion cells and hemangiomatous vessels, occurring focally and causing recurrent obstructive symptoms or occult chronic gastrointestinal bleeding. In this paper we report our experience with this tumor.