bluuebunny/crossref_metadata_2025 huggingface dataset:
Human immunodeficiency virus type 1 subtype C Gag virus-like particle boost substantially improves the immune response to a subtype C gag DNA vaccine in mice
Authors: Jaffray Ann , Shephard Enid , van Harmelen Joanne , Williamson Carolyn , Williamson Anna-Lise , Rybicki Edward P.
Published: February 2004

Human immunodeficiency virus type 1 (HIV-1) subtype C is the predominant HIV in southern Africa, and is the target of a number of recent vaccine candidates. It has been proposed that a heterologous prime/boost vaccination strategy may result in stronger, broader and more prolonged immune responses. Since HIV-1 Gag Pr55 polyprotein can assemble into virus-like particles (VLPs) which have been shown to induce a strong cellular immune response in animals, we showed that a typical southern African subtype C Pr55 protein expressed in insect cells via recombinant baculovirus could form VLPs. We then used the baculovirus-produced VLPs as a boost to a subtype C HIV-1 gag DNA prime vaccination in mice. This study shows that a low dose of HIV-1 subtype C Gag VLPs can significantly boost the immune response to a single subtype C gag DNA inoculation in mice. These results suggest a possible vaccination regimen for humans.
Isolation and identification of a novel human parechovirus
Authors: Ito Miyabi , Yamashita Teruo , Tsuzuki Hideaki , Takeda Naokazu , Sakae Kenji
Published: February 2004

A cytopathic agent (A308/99) was isolated using Vero cells from a stool specimen of a 1-year-old patient with transient paralysis. The agent was approximately 28 nm in diameter with a distinct ultrastructure resembling the virus particle of an enterovirus. It could not be neutralized by antisera against human picornaviruses such as human enterovirus, Aichi virus or human parechovirus. The virion contained three capsid proteins with molecular masses of 38, 30·3 and 30 kDa. Determination of the complete nucleotide sequence of A308/99 revealed that the nucleotide and deduced amino acid sequences were closely related to those of human parechoviruses. When 11 regions encoding the structural and non-structural proteins were compared, A308/99 had between 75 and 97 % and 73 and 97 % nucleotide identity with human parechovirus type 1 (HPeV-1) and type 2 (HPeV-2), respectively. The most distinctive divergence was seen in VP1, which had 74·5 % and 73·1 % nucleotide identity with HPeV-1 and HPeV-2, respectively. Viruses related to A308/99 were also isolated from three patients with gastroenteritis, exanthema or respiratory illnesses. A308/99 and these other three isolates had no arginine–glycine–aspartic acid (RGD) motif, which is located near the C terminus of VP1 in HPeV-1 and HPeV-2. A seroepidemiological study revealed that the prevalence of A308/99 antibodies was low (15 %) among infants but became higher with age, reaching more than 80 % by 30 years of age. These observations indicate that A308/99 is genetically close to, but serologically and genetically distinct from, HPeV-1 and HPeV-2 and accordingly can be classified as third serotype of human parechovirus.
Recombination in uveitis-causing enterovirus strains
Authors: Lukashev A. N. , Lashkevich V. A. , Koroleva G. A. , Ilonen J. , Hinkkanen A. E.
Published: February 2004

The complete nucleotide sequences of three human echovirus (EV) 11 strains and one EV19 strain, all of which caused outbreaks of enterovirus uveitis (EU), a new infant disease first identified in 1980 in Siberia, were determined. One EV11 strain which caused an outbreak of sepsis-like disease in Hungary was also sequenced. All four EV11 strains were mosaic recombinants of the prototype EV11 strain Gregory, with their non-structural coding regions and 5′ NTRs being more similar to other prototype enteroviruses (EV1, EV9). However, this finding is probably a feature of all circulating enterovirus strains and may not be related to their altered virulence. A full genome sequence comparison of the three subtypes of EU-causing strains excludes the role of recent recombination in their emergence, and points to their independent emergence.
Replication-incompetent virions of Japanese encephalitis virus trigger neuronal cell death by oxidative stress in a culture system
Authors: Lin Ren-Jye , Liao Ching-Len , Lin Yi-Ling
Published: February 2004

It has been shown that replication of the Japanese encephalitis virus (JEV) can trigger infected cells to undergo apoptosis. In the present study, it is further demonstrated that replication-incompetent virions of JEV, obtained by short-wavelength ultraviolet (UV) irradiation, could also induce host-cell death. It was found that UV-inactivated JEV (UV-JEV) caused cell death in neuronal cells such as mouse neuroblastoma N18 and human neuronal NT-2 cells, but not in non-neuronal baby hamster kidney BHK-21 fibroblast or human cervical HeLa cells. Only actively growing, but not growth-arrested, cells were susceptible to the cytotoxic effects of UV-JEV. Killing of UV-JEV-infected N18 cells could be antagonized by co-infection with live, infectious JEV, suggesting that virions of UV-JEV might engage an as-yet-unidentified receptor-mediated death-signalling pathway. Characteristically, mitochondrial alterations were evident in UV-JEV-infected N18 cells, as revealed by electron microscopy and a loss of membrane potential. N18 cells infected by UV-JEV induced generation of reactive oxygen species (ROS) as well as the activation of nuclear factor kappa B (NF-κB), and the addition of anti-oxidants or specific NF-κB inhibitors to the media greatly reduced the cytotoxicity of UV-JEV. Together, the results presented here suggest that replication-incompetent UV-JEV damages actively growing neuronal cells through a ROS-mediated pathway.
Evolution of wild-type 1 poliovirus in two healthy siblings excreting the virus over a period of 6 months
Authors: Hovi Tapani , Lindholm Noora , Savolainen Carita , Stenvik Mirja , Burns Cara
Published: February 2004

Wild-type 1 poliovirus (wtPV1) strains were isolated from two young healthy brothers shortly after arrival in Finland from Somalia in 1993. Twelve (sibling A) and 18 (sibling B) specimens collected over a period of more than 6 months yielded wtPV1. Partial sequences obtained from the one and two earliest isolates from sibling A and B, respectively, were nearly identical, differing from each other by only one or two nucleotides. Subsequently, the virus evolved separately in both siblings so that maximal differences between strains derived from a given subject peaked at 2·2 % for sibling A, at 1·5 % for sibling B and at 2·5 % between the two siblings in the VP1-coding part of the genome. All substitutions in the 150 nt VP1–2A junction region were synonymous, whereas as many as eight of the 31 variable positions in the remaining VP1-coding region encoded amino acid replacements in at least one strain. Probable structural locations of the variable amino acid positions were mapped to the published PV1/Mahoney structural model. Most of the substitutions occurred around the fivefold axis in motifs that are known to be or suspected to be targets of neutralizing antibodies. We suggest that the striking genetic divergence observed between the strains was based on a combination of bottleneck transmission events and antigenic drift during the prolonged period of poliovirus replication.
Prion protein gene polymorphisms in healthy and scrapie-affected sheep in Greece
Authors: Billinis Charalambos , Psychas Vassilios , Leontides Leonidas , Spyrou Vassiliki , Argyroudis Stamatis , Vlemmas Ioannis , Leontides Sotirios , Sklaviadis Theodoros , Papadopoulos Orestis
Published: February 2004

A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case–control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).
Extent of reduction of foot-and-mouth disease virus RNA load in oesophageal–pharyngeal fluid after peak levels may be a critical determinant of virus persistence in infected cattle
Authors: Zhang Zhidong , Murphy Ciara , Quan Melvyn , Knight Jeanette , Alexandersen Soren
Published: February 2004

To investigate whether foot-and-mouth disease virus (FMDV) RNA loads in oesophageal–pharyngeal fluid (OP-fluid) in the early course of infection is related to the outcome of virus persistence, viral RNA in OP-fluid samples from cattle experimentally infected with FMDV type O was quantitatively analysed by using a quantitative real-time RT-PCR. Viral RNA was detected within 24 h post-infection (p.i.) in all infected animals. Rapid virus replication led to peak levels of viral RNA load by 30–53 h p.i., and then the load declined at various rates. In some animals (n=12, so-called non-carriers) viral RNA became undetectable between 7 and 18 days p.i. In contrast, in persistently infected animals (n=12, so-called carriers) viral RNA persisted in OP-fluid samples at detectable levels beyond 28 days p.i. Analysis of early viral decay/clearance and virus clearance half-life in OP-fluid samples showed that the extent of reduction of viral RNA in OP-fluid samples immediately following peak levels is a critical determinant of the outcome of FMDV persistence.
Dual targeting of the human cytomegalovirus UL37 exon 1 protein during permissive infection
Authors: Mavinakere Manohara S. , Colberg-Poley Anamaris M.
Published: February 2004

The human cytomegalovirus (HCMV) UL37 immediate-early (IE) gene minimally encodes three protein isoforms that share NH2-terminal sequences. The predominant UL37 isoform detected during HCMV infection was the UL37 exon 1 protein (pUL37x1), which was produced from IE and, more abundantly, through late times of infection. pUL37x1 was localized in both the endoplasmic reticulum (ER) and mitochondria in infected cells. To determine which UL37x1 NH2-terminal residues serve as ER and mitochondrial targeting signals, we examined the subcellular localization of two deletion mutants. pUL37x1Δ2–23, which lacks the hydrophobic leader, is neither translocated into the ER nor imported mitochondrially; conversely, pUL37x1Δ23–34, lacking the juxtaposed basic residues, was translocated into the ER but only imported weakly into mitochondria. These studies show for the first time the temporal production and localization of pUL37x1 during HCMV infection. The trafficking patterns of mutants suggest that the pUL37x1 targeting signal to ER and mitochondria is bipartite.
Absence of tumour necrosis factor facilitates primary and recurrent herpes simplex virus-1 infections
Authors: Minagawa Hiroko , Hashimoto Koji , Yanagi Yusuke
Published: February 2004

Tumour necrosis factor (TNF) is an important cytokine in the innate immune response against various infections, including herpes simplex virus (HSV) infection. It has recently become a molecular target of anti-cytokine treatment in certain inflammatory diseases. TNF depletion resulted in a more rapid emergence of infectious HSV-1 in the explant cultures of latently infected trigeminal ganglia (TG), compared with controls. To further evaluate the importance of TNF in the host's defence responses against HSV-1, TNF-knockout mice were challenged via scarified cornea. These mice were more susceptible to primary acute corneal HSV-1 infection than controls, as manifested by an increased mortality rate and higher infectious virus titres in the eyes and TG, indicating that TNF is critical for defence during acute HSV infection. These results imply that the administration of anti-inflammatory TNF antagonists might facilitate the propagation of infectious HSV, resulting in an exacerbation of primary and recurrent acute lesions.
Genotype C of hepatitis B virus can be classified into at least two subgroups
Authors: Huy Tran Thien-Tuan , Ushijima Hiroshi , Quang Vo Xuan , Win Khin Maung , Luengrojanakul Pairoj , Kikuchi Kaoru , Sata Tetsutaro , Abe Kenji
Published: February 2004

A genomic characterization of hepatitis B virus (HBV) was done for 56 pre-S1/pre-S2 genes and 10 full-length HBV genotype C isolates from five Asian countries. Phylogenetic analysis of the pre-S1/pre-S2 genes revealed two major groups within genotype C: one for isolates from southeast Asia including Vietnam, Myanmar and Thailand (named HBV/C1) and the other for isolates from Far East Asia including Japan, Korea and China (named HBV/C2). This finding was confirmed by phylogenetic analysis based on the full-length sequence of 32 HBV genotype C isolates, including 22 from database entries. Two isolates from Okinawa, the island off the southern end of Japan, formed a different branch. Specific amino acid sequence changes were identified in the large S protein (amino acids 51, 54, 60, 62 and 73) and P protein (amino acids 231, 233, 236, 248, 252 and 304). Our results indicate that genotype C of HBV can be classified into at least two subgroups.
A conserved basic loop in hepatitis C virus p7 protein is required for amantadine-sensitive ion channel activity in mammalian cells but is dispensable for localization to mitochondria
Authors: Griffin Stephen D. C. , Harvey Ruth , Clarke Dean S. , Barclay Wendy S. , Harris Mark , Rowlands David J.
Published: February 2004

We previously identified the function of the hepatitis C virus (HCV) p7 protein as an ion channel in artificial lipid bilayers and demonstrated that this in vitro activity is inhibited by amantadine. Here we show that the ion channel activity of HCV p7 expressed in mammalian cells can substitute for that of influenza virus M2 in a cell-based assay. This was also the case for the p7 from the related virus, bovine viral diarrhoea virus (BVDV). Moreover, amantadine was shown to abrogate HCV p7 function in this assay at a concentration that specifically inhibits M2. Mutation of a conserved basic loop located between the two predicted trans-membrane alpha helices rendered HCV p7 non-functional as an ion channel. The intracellular localization of p7 was unaffected by this mutation and was found to overlap significantly with membranes associated with mitochondria. Demonstration of p7 ion channel activity in cellular membranes and its inhibition by amantadine affirm the protein as a target for future anti-viral chemotherapy.
The Effect of Inflation on Government Revenue and Expenditure: The Case of the Islamic Republic of Iran
Authors: Alavirad Abbas
Published: February 2004

This paper provides an empirical analysis of the government budget deficit, money supply growth and inflation in the Islamic Republic of Iran. We use a simultaneous equation system to examine the effect of inflation on government revenue and expenditure, using quarterly data. Our major finding has been that the government budget deficit will increase in the inflationary condition. In addition, the deficit will increase money supply and this tends to increase inflation in Iran.
Seasonal dynamics of direct and indirect condition indices in relation to energy allocation in largemouth bass Micropterus salmoides (Lacèpede)
Authors: Brown M. L. , Murphy B. R.
Published: February 2004

Abstract  – We investigated several indices (relative weight, mesentery fat, hepatosomatic index, gonadosomatic index, muscle index and visceral somatic index) of largemouth bass Micropterus salmoides (Lacèpede) as a function of season, gender and length to determine how these indices changed with energy content. Length was a significant influence in all comparisons of monthly index measurements, with the exception of the muscle index. All indices, except for the gonadosomatic index, showed few significant differences between sexes for all length groups. Generally, index values were associated with seasonal changes in prey relative abundance and temperature, and the reproductive cycle in mature fish. Total body energy was most influenced by muscle energy stores. Hepatic and gonadal energy of large, sexually mature females decreased considerably with the onset of spawning. Muscle and hepatic energy, to a lesser extent, supported gonadal development. Muscle index values closely tracked seasonal changes in muscle energy, thus providing a useful tool for evaluating seasonal growth and energy patterns. The hepatosomatic index responded to changes in body condition of small and medium fish until June, after which further increases in plumpness caused the utility of this index to decrease. The mesentery fat index provided a useful measure of stored energy available for long‐term needs, relative to other energy depots. Seasonal dynamics of invasive condition indices must be considered in light of rapid increases or decreases in body weight (e.g., spawning loss of gametes).
Habitat use by the African anabantid fish Ctenopoma muriei: implications for costs of air breathing
Authors: Randle A. M. , Chapman L. J.
Published: February 2004

Abstract –  This study examined whether habitat use by the African anabantid fish Ctenopoma muriei is consistent with predictions relating to two costs of air breathing (surface travel and aerial predation). We predicted that C. muriei would be most abundant in habitats that minimize potential costs of air breathing. We quantified environmental correlates of C. muriei abundance within a series of wetland lagoons in Uganda and found that C. muriei was more abundant in shallow waters of the deepest lagoon, in vegetated habitats, and at a depth 15–30 cm below the surface. These locations may minimize cost of travel to the surface for aerial respiration, while maintaining a reasonable distance from the surface to avoid detection by aerial predators. In addition, we experimentally tested effectiveness of vegetative cover in reducing mortality of C. muriei by aerial predators (pied kingfishers) using enclosures with and without vegetation cover in a swamp pool. We found that the presence of vegetative cover significantly reduced susceptibility of C. muriei to predation by pied kingfishers when dissolved oxygen in the water was low and air‐breathing frequency was potentially high.
Habitat use and population structure of four native minnows (family Cyprinidae) in the upper Missouri and lower Yellowstone rivers, North Dakota (USA)
Authors: Welker T. L. , Scarnecchia D. L.
Published: February 2004

Abstract –  In 1997 and 1998, sampling was conducted on the Missouri and Yellowstone rivers, North Dakota, to obtain information on the distribution, abundance, and habitat use of the flathead chub (Platygobio gracilis Richardson), sicklefin chub (Macrhybopsis meeki Jordan & Evermann), sturgeon chub (Macrhybopsis gelida Girard), and western silvery minnow (Hybognathus argyritis Girard), four declining fish species (family Cyprinidae) native to the Missouri River basin, USA. The study area consisted of four distinct river segments near the confluence of the Missouri and Yellowstone rivers – three moderately altered segments that were influenced by a main‐stem dam and one quasi‐natural segment. One moderately altered segment was located at the confluence of the two rivers (mixing‐zone segment (MZS)). The other two moderately altered segments were in the Missouri River adjacent to the MZS and extended up‐river (above‐confluence segment (ACS)) and down‐river (below‐confluence segment (BCS)) from this segment. The quasi‐natural segment (Yellowstone River segment (YRS)) extended up‐river from the MZS in the Yellowstone River. Catch rates with the trawl for sicklefin chub and sturgeon chub and catch rates with the bag seine for flathead chub and western silvery minnow were highest in the BCS and YRS. Most sicklefin and sturgeon chubs were captured in the deep, high‐velocity main channel habitat with the trawl (sicklefin chub, 97%; sturgeon chub, 85%), whereas most flathead chub and western silvery minnow were captured in the shallow, low‐velocity channel border habitat with the bag seine (flathead chub, 99%; western silvery minnow, 98%). Best‐fit regression models correctly predicted the presence or absence of sicklefin chub, flathead chub, and western silvery minnow more than 80% of the time. Sturgeon chub presence and absence were predicted correctly 55% of the time. Best‐fit regression models fit to fish number data for flathead chub, sicklefin chub, and sturgeon chub and fish catch‐per‐unit‐effort (CPUE) data for flathead chub also provided good fits, with R2 values ranging from 0.32 to 0.55 (P < 0.0001). The higher density and catch of the four native minnows in the YRS and BCS suggest that these two segments are better habitat than the ACS and MZS.
Vertical distribution patterns of zooplanktivorous fish in a shallow, eutrophic lake, mediated by water transparency
Authors: Mous P. J. , Van Densen W. L. T. , Machiels M. A. M.
Published: February 2004

Abstract –  The vertical distribution pattern (VDP) of fish at shallow sites in eutrophic lake – Lake IJssel, the Netherlands – as affected by water transparency, was examined. The pattern was assessed by pair trawling at three depths and by hydroacoustics from June to August. Water transparency was estimated by light intensity and Secchi depth measurements. Hydroacoustics showed that fish were more dispersed at night than during the day. European smelt, (Osmerus eperlanus) and young‐of‐the‐year Eurasian perch (Perca fluviatilis) comprised 97% of the fish in numbers. European smelt concentrated near the surface, at light intensities of >200 µW · cm−2 when the water was turbid, and near the bottom when the water was clear (Secchi depth > 1.2 m). We concluded that low water transparency over‐ruled the light sensitivity of European smelt, and caused European smelt to stay higher in the water column.
ERK activation by mechanical strain is regulated by the small G proteins rac‐1 and rhoA
Authors: Laboureau Julien , Dubertret Louis , Lebreton‐De Coster Corinne , Coulomb Bernard
Published: February 2004

Abstract: Physical forces play an important role in regulating cell functions. We applied mechanical strain to human fibroblasts by magnetic attraction of superparamagnetic arginine‐glycine‐aspartic acid (RGD)‐coated beads. We confirmed that the MAP kinases Erk and p38 are activated by mechanical strain, and went further by demonstrating the activation of Elk‐1 by mechanical strain, mainly through a MEK‐Erk pathway. Transfection of a dominant negative form of the G protein rac‐1 (rac T17N), and inhibition of PI3K, an effector of rac‐1, efficiently prevented Elk‐1 activation by mechanical forces. Transfection with C3 transferase, known to inhibit rhoA, and inhibition of rock (a downstream effector of rhoA), gave similar results. However, contrary to the active form of rhoA (rho G14V), transfection of the active form of rac‐1 (rac G12V) induced Elk activation and mimicked the effects of mechanical strain. These results point out that the two small G proteins rhoA and rac‐1 participate in cell sensitivity to mechanical strain and lead to the modulation of the Erk pathway.
Antipsoriatic drug anthralin induces EGF receptor phosphorylation in keratinocytes: requirement for H2O2 generation
Authors: Peus Dominik , Beyerle Astrid , Vasa Mariuca , Pott Markus , Meves Alexander , Pittelkow Mark R.
Published: February 2004

Abstract: Even though anthralin is a well‐established topical therapeutic agent for psoriasis, little is known about its effects and biochemical mechanisms of signal transduction. In contrast to a previous report, we found that anthralin induced time‐ and concentration‐dependent phosphorylation of epidermal growth factor receptor in primary human keratinocytes. Four lines of evidence show that this process is mediated by reactive oxygen species. First, we found that anthralin induces time‐dependent generation of H2O2. Second, there is a correlation between a time‐dependent increase in anthralin‐induced epidermal growth factor receptor phosphorylation and H2O2 generation. Third, the structurally different antioxidants n‐propyl gallate and N‐acetylcysteine inhibited epidermal growth factor receptor phosphorylation induced by anthralin. Fourth, overexpression of catalase inhibited this process. The epidermal growth factor receptor‐specific tyrosine kinase inhibitor PD153035 abrogated anthralin‐induced epidermal growth factor receptor phosphorylation and activation of extracellular‐regulated kinase 1/2. These findings establish the following sequence of events: (1) H2O2 generation, (2) epidermal growth factor receptor phosphorylation, and (3) extracellular‐regulated kinase activation. Our data identify anthralin‐induced reactive oxygen species and, more specifically, H2O2 as an important upstream mediator required for ligand‐independent epidermal growth factor receptor phosphorylation and downstream signaling.
Analysis of APAF‐1 expression in human cutaneous melanoma progression
Authors: Baldi Alfonso , Santini Daniele , Russo Patrizia , Catricalà Caterina , Amantea Ada , Picardo Mauro , Tatangelo Fabiana , Botti Gerardo , Dragonetti Emanuele , Murace Raffaele , Tonini Giuseppe , Natali Pier Giorgio , Baldi Feliciano , Paggi Marco G.
Published: February 2004

Abstract: APAF‐1 plays a pivotal role in mitochondria‐dependent apoptosis, binding to cytochrome c and favoring activation of caspase‐9. It has been shown that epigenetic silencing of the APAF‐1 gene is a common event in several metastatic melanoma cells in vitro. We determined, by Western blot, variation in the level of expression of APAF‐1 in several human melanoma cell lines and, by immunohistochemistry, in a group of 106 histological samples including benign and malignant melanocytic lesions. We observed APAF‐1 down‐regulation or loss of expression in two metastatic melanoma cell lines, compared to primary melanoma cell lines. The immunohistochemical analysis revealed a significant difference in APAF‐1 staining between nevi and melanomas. In addition, we found a significant negative correlation between APAF‐1 expression level and tumor thickness and between primary melanomas and metastases. We conclude that loss of APAF‐1 expression can be considered as an indicator of malignant transformation in melanoma.
Comparative expression of vascular endothelial growth factor family members, VEGF‐B, ‐C and ‐D, by normal human keratinocytes and fibroblasts
Authors: Trompezinski S. , Berthier‐Vergnes O. , Denis A. , Schmitt D. , Viac J.
Published: February 2004

Abstract:  The vascular endothelial growth factor (VEGF) family includes the related polypeptides VEGF‐B, ‐C and ‐D, which contribute to endothelial and lymphatic vessel development. The parental VEGF molecule, VEGF‐A, has been widely described in the skin, but the other members of the VEGF family have not yet been reported.

The aim of our study was to determine whether the two main skin cells, keratinocytes and fibroblasts, expressed VEGF‐B, ‐C and ‐D in basal condition and after stimulation by either growth factors or the pro‐inflammatory cytokine tumour necrosis factor‐α (TNF‐α). Reverse‐transcription polymerase chain reaction (RT‐PCR) analysis on cultured normal human keratinocytes (NHKs) and normal human fibroblasts (NHFs) allowed the detection of different levels of VEGF‐B, ‐C and ‐D mRNA, in both cell types with similar RT‐PCR products in the skin cells. A semi‐quantitative evaluation of the VEGF family proteins by dot blot, using the different human recombinant VEGFs, showed different levels of VEGF‐B, ‐C and ‐D, in NHKs and NHFs. After cell stimulation by growth factors (epidermal growth factor (EGF) and transforming growth factor‐β1 (TGF‐β1) for NHKs and NHFs, respectively), a significant up‐regulation of the VEGF family member proteins was observed in NHFs but not in NHKs. Conversely, TNF‐α did not exert a significant effect. However, we could not detect any transcriptional modification in stimulated cells, whatever the stimulation duration. The addition of cycloheximide to the cell cultures strongly inhibited the increase of VEGF proteins in TGF‐β1‐stimulated NHFs. Taken together, the results underline the major role played by NHFs in the elaboration of the VEGF family proteins known to regulate wound healing, chronic inflammation and tumour angiogenesis and lymphangiogenesis.
A very mild form of non‐Herlitz junctional epidermolysis bullosa: BP180 rescue by outsplicing of mutated exon 30 coding for the COL15 domain
Authors: Pasmooij A. M. G. , Van Zalen S. , Nijenhuis A. M. , Kloosterhuis A. J. , Zuiderveen J. , Jonkman M. F. , Pas H. H.
Published: February 2004

Abstract: Mutations in the gene COL17A1 cause non‐Herlitz junctional epidermolysis bullosa. Here, we describe a patient who, despite two heterozygous mutations in COL17A1, has an extremely mild form of the disease missing most of the characteristic clinical features. DNA analysis revealed a frame‐shift mutation 3432delT and a nonsense mutation 2356C→T (Q751X). cDNA analysis showed that the deleterious effect of the latter mutation was skirted by deleting the premature termination codon containing exon 30. In this way, the reading frame was restored, resulting in a 36 nucleotides shorter mRNA transcript. Immunoblot analysis showed expression of the 180‐kDa bullous pemphigoid antigen (BP180) with a slightly higher SDS‐PAGE mobility, in line with the deletion of 12 amino acids from the COL15 domain. Immunofluorescence of skin sections showed diminished, but correctly localised expression of BP180, and this, in concert with the mild clinical phenotype, suggests that this COL15 mutated BP180 is still partly functional.
Tumor necrosis factor‐α gene expression and release in cultured human dermal microvascular endothelial cells
Authors: Hoffmann Georg , Schloesser Michaela , Czechowski Monika , Schobersberger Wolfgang , Fürhapter Christine , Sepp Norbert
Published: February 2004

Abstract:  Endothelial dysfunctions in the microcirculation are a common finding in the course of inflammatory disorders. These are, at least in part, mediated by endogenous agonists, e.g. tumor necrosis factor‐α (TNF‐α). As TNF‐α mostly acts in an autocrine or paracrine fashion, it was tempting to speculate that microvascular endothelial cells synthesize and release this cytokine upon appropriate stimulation. In the present study, human dermal microvascular endothelial cells (HDMECs) expressed the TNF‐α gene following incubation with interleukin‐1β (IL‐1β), lipopolysaccharids (LPSs), as well as a combination of IL‐1β, LPSs, and interferon‐γ (IFN‐γ), while IFN‐γ failed to exert an effect on TNF‐α gene expression when given as a single stimulus. Transcription of the TNF‐α gene was accompanied by an increase in TNF‐α protein secretion into the cellular supernatant. As HDMECs were found to be a target of TNF‐α, production of this cytokine by HDMECs may result in an autocrine activation loop that contributes to the deterioration of microcirculatory functions in infectious diseases and inflammatory skin disorders.
Acute hemorrhagic edema of infancy
Authors: PoyrazoĞlu Hakan M , Per Hüseyin , Gündüz Zübeyde , DüŞünsel Ruhan , Arslan Duran , Narİn Nazmi , GümüŞ Hakan
Published: February 2004

Abstract
Background
: Acute hemorrhagic edema of infancy (AHEI) is a leukocytoclastic vasculitis of infants. Even though the cutaneous findings are dramatic in both appearance and rapidity of onset, the prognosis is benign.

Methods
: We report on eight patients with AHEI evaluated from the viewpoints of age, sex, history of infection, drug administration or immunization before the onset of skin lesions, laboratory findings, cutaneous findings, cutaneous biopsy and treatment.

Results
: The ages of the patients, six of whom were male, ranged between 7 and 27 months. Five episodes in our patients occured in winter. Six patients (75%) had a history of recent infection, drug administration or immunization. All patients had fever, peripheral edema and large, purpuric, annular or targetoid plaques on the face and extremities. Two patients had mucosal involvement (conjunctiva and palatum molle). One patient with severe genital involvement had trunk involvement. Hematologic and immunologic tests were not diagnostic in any patient. Leukocytoclastic vasculitis was demonstrated in four patients for whom a cutaneous biopsy was performed. Complete recovery time ranged between 4 and 7 days.

Conclusions
: We believe that AHEI is a benign disorder, despite its dramatic appearance. We want to point out that mucosal and trunk involvement may also occur in AHEI.
Comparison of ibuprofen and indomethacin therapy for patent ductus arteriosus in preterm infants
Authors: Su Pen‐Hua , Chen Jia‐Yuh , Su Chi‐Ming , Huang Tzu‐Ching , Lee Hong‐Shen
Published: February 2004

Background
: Patent ductus arteriosus (PDA) is commonly found in very low‐birthweight (VLBW) infants. The presence of respiratory distress syndrome (RDS) is also associated with increased frequency of significant PDA. Intravenous indomethacin has been used to treat and to prevent PDA in premature infants since 1976. However, concern remains regarding the safety of indomethacin, which affects renal, gastrointestinal and cerebral perfusion. Intravenous ibuprofen has recently been used to treat and to prevent PDA premature infants with PDA without reducing cerebral blood flow or affecting intestinal or renal hemodynamics. The aim of the present study is to compare intravenous ibuprofen and indomethacin with regard to efficacy and safety for the early treatment of PDA in preterm infants.

Methods
: A total of 63 preterm infants with RDS who had a birthweight of ≤1500 g and gestational age of ≤32 weeks, were enrolled in the present study. All patients were treated with nasal continuous positive airway pressure with additional oxygen supply in inspired air>30%, or with mechanical ventilation. The patients’ serum platelet counts were>100 000/uL, and serum creatinine values were <1.5 mg/dL. There were no 3−4 grade intraventricular hemorrhages before randomization, and all patients were aged 2−7 days and had echo­cardio­graphic evidence of significant PDA. Patients were randomized into two groups: the first group of neonates (group A, n = 32) received intravenous ibuprofen lysine 10 mg/kg, followed by 5 mg/kg after 24 and 48 h; the second group (group B, n = 31) received intravenous indomethacin 0.2 mg/kg every 12 h for three doses.

Results
: Patent ductus arteriosus closed in 27 patients from the ibuprofen group (84.4%) and in 25 patients from the indomethacin group (80.6%). PDA reopened in three patients from the ibuprofen group (9.4%) and in three patients from the indomethacin group (9.7%). One patient in the ibuprofen group and two patients in the indomethacin group required ductal ligation. Serum creatinine and blood urea nitrogen (BUN) concentrations were lower in the ibuprofen group than in the indomethacin group. Urine output and creatinine clearance values were higher in the ibuprofen group than in the indomethacin group.

Conclusions
: Ibuprofen therapy is as efficacious as indomethacin for the treatment of PDA in preterm infants. Infants treated with ibuprofen have higher creatinine clearance and urine output and lower serum creatinine and BUN values than infants treated with indomethacin.
Tolmetin and salicylate therapy in acute rheumatic fever: Comparison of clinical efficacy and side‐effects
Authors: Karademir Selmin , OĞuz Deniz , Şenocak Filiz , Öcal Burhan , Karakurt CemŞit , Çabuk Feryal
Published: February 2004

Abstract
Background
: The arthritis of rheumatic fever is very responsive to treatment with salicylates, but there are many adverse reactions, especially hepatotoxicity, due to aspirin (acetylsalicylic acid) therapy. These side‐effects change the course and duration of rheumatic fever. Other non‐steroidal anti‐inflammatory drugs may be equally effective, although no reports are available.

Methods
: We studied 72 patients with rheumatic fever who were admitted to Dr Sami Ulus Children's Hospital between 1995 and 1999. Twenty patients with arthritis were treated with tolmetin (25 mg/kg per day; group I) and 52 patients with arthritis and/or mild carditis were put on aspirin therapy (75−100 mg/kg per day) for 4−6 weeks (group II). Arthritis had disappeared at the same time in both the aspirin and tolmetin groups (P = 0.675).

Results
: The erythrocyte sedimentation rates of patients upon admission, at the first week and at the end of therapy were not different in the two groups (P > 0.05). No adverse effect of tolmetin therapy was observed, whereas side‐effects of salicylate were observed in 19 patients (36.5%) in the aspirin group. Hepatotoxicity, gastric irritation and salicylism were found in 16, four and three patients, respectively. Renal toxicity and Reye syndrome were not demonstrated. Because of these side‐effects of aspirin, therapy had to be stopped for 10−20 days and the duration of hospitalization in this group was lengthened unnecessarily.

Conclusion
: Tolmetin was safe and effective treatment for arthritic rheumatic fever patients without carditis. Tolmetin can be used particularly in patients who cannot tolerate aspirin.
Evaluation of change of cerebral circulation by SpO2 in preterm infants with apneic episodes using near infrared spectroscopy
Authors: Yamamoto Akiyo , Yokoyama Naoki , Yonetani Masahiko , Uetani Yoshiyuki , Nakamura Hajime , Nakao Hideto
Published: February 2004

Abstract
Background
: In the neonatal intensive care unit (NICU), hemodynamics in very low‐birthweight infants are generally examined for oxygen saturation (SpO2), heart rate, respiration rate, and blood pressure. The present study examined how changes in cerebral circulation in preterm infants can be evaluated by the SpO2 monitoring method with near infrared spectroscopy (NIRS) to detect the cerebral circulation.

Methods
: The study was conducted in 11 low‐birthweight neonates with a mean weight of 1252 g (940−1948 g), mean post‐conceptional age of 28.9 weeks (28−31 weeks) and in whom a total of 145 apneic episodes were examined. Changes in cerebral circulation at the apneic attack were evaluated by two parameters of ΔHbD (µmol/L) for reduction in cerebral oxygenation and ΔcHb (mL/100 g brain) for variation in cerebral blood volume using the near infrared spectroscopy (NIRS).

Results
: There was a tendency for a reduction in cerebral oxygenation and a change in cerebral blood volume as SpO2 was reduced. In the event of apneic attacks where SpO2 was reduced to <85%, cerebral oxygen saturation was extensively reduced. In addition, cerebral blood volume was also greatly changed when the SpO2 was reduced to <85%, and changed further still when SpO2 was reduced again to ≤75%.

Conclusion
: Reduction in SpO2 (<85%) was suggested to be an effective indication to changes in cerebral circulation. In the case of apneic attacks where SpO2 was ≤85%, the cerebral circulation in preterm low‐birthweight neonates was extensively changed and, therefore, attention should be paid to changes in the concentration of SpO2 when managing apnea of prematurity in NICU.
Effect of tongue brushing on oral malodor in adolescents
Authors: Çiçek Yasin , Orbak Recep , Tezel Adnan , Orbak Zerrin , Erciyas Kamile
Published: February 2004

Abstract
Background
: Halitosis is a common problem in humans, and is a social and psychological handicap for those affected by it. Halitosis has a positive correlation with the accumulation of bacterial plaque in the oral cavity.

Aims
: The aim of the present study was to determine the effect of tongue brushing on oral malodor in adolescents.

Methods
: The subjects of the investigation were 28 adolescents who had oral malodor and whose average age was 16 ± 0.12 years. Subjects were chosen to participate in the study if they had an established routine of oral care, and did not have systemic or periodontal disease or dentures (such as fixed prosthesis or partial and total dentures). The subjects with oral malodor were randomly assigned into two groups. Individuals in the first group received instructions regarding professional procedures of oral care from a periodontist. In addition, individuals in this group had the surface of their tongue cleaned using a hard toothbrush, wetted with 0.12% chlorhexidine gluconate. Individuals in the second group (control) underwent the same professional procedures of oral care executed by the same periodontist, however, they did not have their tongues cleaned. Oral malodor of individuals was then evaluated for all individuals using the organoleptic method. The pH of the subjects tongues were recorded before and after treatment. The results of the oral malodor test were compared between the two groups.

Results
: The results of the study found that oral malodor scores obtained by an organoleptic method was more severe in the second group than the first group, and statistical differences were found between the two groups (P < 0.001).

Conclusion
: These results revealed that accumulation of bacterial plaque on the tongue is an important factor for oral malodor in the adolescents. Oral malodor levels were significantly reduced after cleaning the surface of the tongue. Thus, tongue care shouldn’t be neglected in order to avoid oral malodor.
Physical and mental development of Turkish twins
Authors: Özçakar Z. Birsin , Sahin Figen , Beyazova Ufuk , Soysal Sebnem
Published: February 2004

Abstract
Background
: Multiple gestations are known to bring some risks in the prenatal and natal period, but whether it is a risk for development in childhood is a matter of debate. The aim of the present study was to evaluate physical and mental development of Turkish twins and then to assess whether being a twin poses risks for their development.

Methods
: Fifty‐two twin pairs (104 children) aged between 1 and 5 years were enroled in the study. The control group consisted of 91 singletons, who were matched according to age, sex, gestational week and maternal educational level. The Denver Developmental Screening Test (DDST) was administered to assess cognitive development. Student t‐test and χ2 test were used to compare the two groups.

Results
: The mean age of the mothers of twins did not differ from that of singleton mothers (P > 0.05). Twins were more frequently born in cases of assisted fertilization and were more often born via cesarean section. The birthweights of twins were lower and they were more frequently hospitalized after birth. The breast feeding period was shorter in twins and they had more chronic diseases than singletons (P < 0.05). Mothers of twins needed more help while caring for their infants. Physical development of twins and singletons did not differ statistically (P > 0.05). Twins had more suspect and delayed results in the DDST, especially in the language section of the test.

Conclusion
: Physical and mental development of twins does not differ significantly from singletons except that twins might have a tendency towards slow language acquisition.
Ketotic hypoglycemia in patients with allergic diseases
Authors: Matsubara Tomoyo , Yoshitomi Tomomi , Kaneko Miho , Ichiyama Takashi , Furukawa Susumu
Published: February 2004

Abstract
Background
: Ketotic hypoglycemia is the most common cause of recurrent hypoglycemia in early childhood but its etiology is poorly understood. Elimination diets have been used for the prevention or treatment of some allergic diseases; however, these diets entail a risk to the normal nutrition and growth of children. The present study investigated whether elimination diets are related to the occurrence of ketotic hypoglycemia.

Methods
: The prevalence of allergy treated with elimination diet therapy was retrospectively investigated in 18 patients with ketotic hypoglycemia seen in Yamaguchi University Hospital between January 1995 and September 1999. Data were gathered by reviewing the patients’ medical charts.

Results
: Ten (55.6%) of 18 patients with ketotic hypoglycemia had allergic diseases. Six (60%) of the patients had been treated with strict elimination diets and the others were on incomplete elimination diets for the prevention of allergy. The ketotic hypoglycemia patients with allergic diseases had experienced fewer pre‐existing infectious diseases than those without allergic diseases.

Conclusions
: Patients with allergic diseases treated with elimination diets have a relatively high tendency towards developing ketotic hypoglycemia. They might have a lower capacity to tolerate fasting due to their excessive avoidance of many foods, even during periods when they are not undergoing strict elimination diets.
Acanthosis nigricans is a reliable cutaneous marker of insulin resistance in obese Japanese children
Authors: Yamazaki Hajime , Ito Sueshi , Yoshida Hiroshi
Published: February 2004

Abstract
Background
: Acanthosis nigricans (AN) is a skin condition characterized by darkening and thickening of skin with formation of irregular folds, usually limited to a few specific areas of the body. Recently, AN has been reported to be linked to hyperinsulinemia and obesity. The aim of the present study was to determine whether or not the presence of AN in obese Japanese children is a reliable cutaneous marker.

Methods
: The authors analyzed the clinical characteristics of 439 obese Japanese children (260 boys, 179 girls; mean age 10.1 years; mean percentage overweight 51.9%), who had visited Tsuruoka City Shonai Hospital in 1990−2000. Eighty‐two of the 439 children were examined using an oral glucose tolerance test (OGTT). Of these children, the authors retrospectively studied 16 subjects: eight with AN and eight without AN (age range: 10.8−13.9 years; percentage overweight range: 54.3−97.0%). They were age and percentage obesity‐matched males with normal glucose tolerance during OGTT. Females with normal glucose tolerance during OGTT were excluded from the 16 subjects because the number was too small and children with impaired glucose tolerance or type 2 diabetes during OGTT were also excluded because of glucose toxicity.

Eighty‐two children including the 16 subjects were analyzed at their first visit for the presence or absence of AN on the posterior of the neck, and for characteristics including age, birthweight, body height, bodyweight, percentage overweight, blood pressure, liver function markers serum lipid concentrations, fasting plasma glucose concentrations and insulin concentrations shown by the results of OGTT.

Results
: (1) Children with AN showed significantly more glucose intolerance including impaired glucose tolerance and type 2 diabetes compared with those children without AN, and fasting plasma insulin concentrations were most significantly correlated with the presence of AN.

(2) Insulin resistance based on fasting plasma insulin concentrations was seen in significantly more children with AN than in children without AN, even in age and percentage obesity‐matched subjects with normal glucose tolerance during OGTT.

Conclusions
:  Acanthosis nigricans could be a reliable cutaneous marker of insulin resistance in obese Japanese children.