bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of cytokines on Porphyromonas gingivalis‐induced opsonophagocytosis of a murine macrophage cell line
Authors: Sosroseno Wihaskoro , Herminajeng Endang , Herwiyanti Sri , Ghufron Muhammad
Published: March 2003

A murine macrophage cell line was incubated with opsonized Porphyromonas gingivalis and various concentrations of rIFN‐γ, rIL‐4 and rIL‐10. The number of phagocyted cells were microscopically assessed. The results showed that IFN‐γ upregulated macrophage phagocytosis to this periodontopathogen, and the upregulatory effects was abolished by anti‐IFN‐γ antibodies. Neither IL‐4 nor IL‐10 had any effects on opsonophagocytosis by this cell line. These results suggest that up‐regulatory functions of IFN‐γ on phagocytic activities of macrophages may play a crucial role in the induction of immune response to P. gingivalis.
Restorations with extensive dentin/enamel‐bonded ceramic coverage. A 5‐year follow‐up
Authors: Van Dijken Jan W. V. , Hasselrot Lars , Örmin Annika , Olofsson Anna‐Lena
Published: March 2003

The durability of restorations with extensive dentin/enamel‐bonded posterior partial and complete ceramic coverages were investigated. The effect of luting with a dual‐cured and a self‐cured luting agent was also studied. In 110 patients, 182 ceramic coverages (IPS Empress) were placed. In 58 restorations, Syntac was used in combination with the dual‐cured resin composite Variolink. In the other restorations luted with the chemically cured resin composite Bisfil 2B, 25 were bonded with Gluma, 57 with Allbond 2, and 42 with Syntac. Of the 182 ceramics, 13 (7.1%) were assessed as non‐acceptable after a mean observation period of 4.9 yr (range 4.3–7.5 yr). The reasons for failure were fracture (5), lost restorations (4), secondary caries (3) and endodontic treatment (1). No significant differences in failure rate were seen between the two luting agents or between the three dentin‐bonding agents. Ceramic coverages placed on non‐vital teeth failed in 9.7% of cases (3/31) and on vital teeth in 6.6% (10/151). The success rate of the dentin‐enamel‐bonded ceramic coverages reduces the need for a traditional full‐coverage therapy and/or post or pin(s) and core placement. The technique investigated showed many clinical advantages such as less destruction of healthy tissue, and avoidance of endodontic treatment and/or deep cervical placement of restoration margins.
Dental fear in sexually abused women
Authors: Willumsen Tiril
Published: March 2003

The aim of the present study was to investigate dental fear in women who have been exposed to different kinds of sexual abuse. In a cross‐sectional questionnaire study, 99 sexually abused women were divided into three groups: one group who reported having been exposed to sexual touching (ST); one group who reported intercourse (IC); and one group who reported sexual abuse involving oral penetration (OP). The mean score on dental fear assessments was significantly higher for all groups than for Norwegian women in general. Women in the OP group scored significantly higher than women in other groups on dental fear. The majority of the women reported that they had experienced problems in relation to dental treatment situations. About half of the women in the OP group and one‐third in the other groups reported that they had never considered that there was a relationship between the abuse and their problems with dental treatment situations. Significantly more women in the OP group reported that they had not been aware of the relationship, possibly because the abuse had been repressed. The majority of the women with extreme dental fear had never informed a dentist that they had been sexually abused.
Apoptosis in oral lichen planus
Authors: Neppelberg Evelyn , Johannessen Anne Christine , Jonsson Roland
Published: March 2003

Apoptotic cell death may be a contributory cause of basal cell destruction in oral lichen planus (OLP). Therefore, the purpose of this study was to investigate the rate of apoptosis in OLP and the expression of two proteins (FasR and FasL) regulating this process. Biopsies from 18 patients with histologically diagnosed OLP were investigated, with comparison to normal oral mucosa of healthy persons. For visualisation of DNA fragmentation, the TUNEL method was used. In order to characterise the infiltrating cell population (CD3, CD4, CD8) and expression of FasR and FasL, we used an immunohistochemical technique. The results showed that T cells dominated in the subepithelial cell infiltrate. Within the epithelium the apoptotic cells were confined to the basal cell layer, and more apoptotic cells were seen in areas with basal cell degeneration and atrophic epithelium. There was a prominent expression of FasR/FasL in OLP, with a rather uniform distribution throughout the inflammatory cell infiltrate. In the epithelium, the FasR/FasL expression was more abundant in the basal cell area compared to the suprabasal cell layer. In conclusion, apoptosis within the epithelium is significantly increased in situ in OLP compared to normal oral mucosa, and seems to be related to the epithelial thickness.
Masticatory performance in patients with anterior disk displacement without reduction in comparison with symptom‐free volunteers
Authors: Peroz Ingrid , Tai Sonja
Published: March 2003

Masticatory function can be impaired by craniomandibular disorders. The aim of this study was to assess masticatory performance in patients with an anterior disc displacement (ADD) without reduction. In the experiments, 29 patients and 33 age‐ and gender‐matched volunteers chewed artificial test food for 60 chewing strokes. The collected remains of the test food were filtered, dried, fractionated by a sieving procedure, and weighed. The particle size distribution was then described using a cumulative distribution function. Patients and controls were clinically examined, and patients were asked to complete a pain questionnaire. Comparison with controls, patients showed significantly reduced masticatory performance. Patients that had had a disorder longer than 3 yr tended to display less reduction of their masticatory performance. Neither the treatment methods used, nor restriction of daily life activities or pain intensity were significantly correlated with masticatory performance. Jaw mobility was significantly reduced in patients. More than half of the patients and none of the controls had joint noises and trigger points in the masticatory muscles. Pain was present, in particular, during chewing and maximal opening of the mouth. It was concluded that patients with ADD without reduction have a significantly reduced masticatory performance.
Matrix metalloproteinase levels and gelatinolytic activity in clinically healthy and inflamed human dental pulps
Authors: Gusman Heloisa , Santana Ronaldo B. , Zehnder Matthias
Published: March 2003

The role of matrix metalloproteinases (MMPs) in the breakdown of pulp tissue of teeth with severe caries has not yet been directly elucidated. This study was to determine the levels of selected MMPs and the overall gelatinolytic activity in clinically healthy and inflamed human dental pulps of 29 healthy subjects, aged 10–19 yr. Seventeen pulps were collected from subjects diagnosed with symptomatic pulpitis, and 18 control pulps were obtained from 12 subjects following premolar extraction for orthodontic reasons. The levels of MMP‐1, MMP‐2, MMP‐3 and MMP‐9 were determined with enzyme‐linked immunosorbent assay. Densitometric analysis of gelatin zymograms was used to assay gelatinolytic activity in pulp supernatants. The MMP‐1 levels were below the detection limit for both groups. Levels of MMP‐2 and MMP‐3 were significantly lower in symptomatic vs. clinically healthy pulps. In contrast, levels of MMP‐9 in inflamed pulps were significantly higher than those recorded in clinically normal pulps. The overall gelatinolytic activity was elevated in inflamed pulps compared with healthy counterparts. Further, the gelatinolytic activity was positively correlated with MMP‐9 levels. The data obtained suggest a key role of MMP‐9 in the breakdown of inflamed human dental pulp tissue.
Possible Roles for Products of Polymorphic MHC and Linked Olfactory Receptor Genes during Selection Processes in Reproduction*
Authors: ZIEGLER ANDREAS , DOHR GOTTFRIED , UCHANSKA‐ZIEGLER BARBARA
Published: March 2003

PROBLEM: Polymorphic genes of the human major histocompatibility complex [MHC; human leukocyte antigen (HLA)] are probably important in determining resistance to parasites and avoidance of inbreeding. We investigated whether HLA‐associated sexual selection could also involve HLA‐linked olfactory receptor (OR) genes, which might not only participate in olfaction‐guided mate choice, but also in selection processes within the testis.

METHOD OF STUDY: The testicular expression status of HLA class I molecules (by immunohistology) and HLA‐linked OR genes (by transcriptional analysis) was determined.

RESULTS: Various HLA class I heavy chains, but not β2‐microglobulin (β2m), were expressed, mainly at the spermatocyte I stage. Of 17 HLA‐linked OR genes analyzed, eight were found to be transcribed in the testis. They exhibited varying numbers of 5′‐ or 3′‐non‐coding exons as well as differential splicing.

CONCLUSIONS: We suggest that testis‐expressed polymorphic HLA and OR proteins are functionally connected and serve the selection of spermatozoa, enabling them to distinguish `self' from `non‐self' [the sperm‐receptor‐selection (SRS) hypothesis].
Endometriosis in Reproductive Immunology
Authors: ULC̆OVÁ‐GALLOVÁ ZDENKA , BOUŠE VLADIMÍR , ŠVÁBEK LADISLAV , TUREK JIR̆I , ROKYTA ZDENĔK
Published: March 2003

PROBLEM: Endometriosis is suggested to represent an autoimmune disorder, but what is the prevalence of autoantibodies to antigens relevant to reproduction?

METHOD OF STUDY: The humoral immune response to the women with endometriosis (stage I–II: 261 women; stage III–IV: 62 women) in serum and in peritoneal fluid was investigated compared with 101 healthy women. Enzyme‐linked immunosorbent assay (ELISA) was used in all the women for the detection of seven antiphospholipid antibodies [antiphospholipid antibodies (aPLs) against cardiolipin, L‐phosphatidyl (ph)‐serine, ph‐glycerol, ph‐inositol, ph‐ethanolamine, phosphatidic (ph)‐acid and against β2‐glycoprotein I] of class IgG, IgA, and IgM. A passive haemmagglutination method and ELISA (BioGen) was used for assessment of antizona pellucida antibodies (aZP), tray agglutination test (TAT) and indirect mixed anti‐imunoglobulin reaction test (MAR‐test) for the determination of sperm antibody levels.

RESULTS: Endometriosis I–II were associated with higher serum and peritoneal fluid levels of aPLs against inositol, cardiolipin, ethanolamine, and β2‐glycoprotein I. Forty percent of patients were positive for aZPA.

CONCLUSIONS: Patients with lesions of endometriosis stage I–II had more autoantibodies than those with stage III–IV, and may be immunologically more active. This result may be significant for future treatments such as in vitro fertilization and embryo transfer.
Transfer of Prostasomal CD59 to CD59‐Deficient Red Blood Cells Results in Protection against Complement‐Mediated Hemolysis
Authors: BABIKER ADIL A. , RONQUIST GUNNAR , NILSSON ULF R. , NILSSON BO
Published: March 2003

PROBLEM: Prostasomes isolated from human seminal plasma have complement regulatory properties because of their content of CD59, a glycosylphosphatidylinositol (GPI)‐anchored protein. We investigated a functional role of prostasomes by the possibility of transferring CD59 from prostasomes to rabbit erythrocytes (RE) and human erythrocytes obtained from patients with paroxysmal nocturnal hemoglobinuria (PNH), both types of cells lacking CD59.

METHOD OF STUDY: We used the assay of hemolytic activity of the alternative pathway of the complement system to compare the liability of the erythrocytes to hemolysis by the complement system with and without pre‐incubation with prostasomes. CD59 gained by the RE and PNH erythrocytes was established by flow cytometry. The effect of phosphatidylinositol phospholipase C (PIPLC) on the GPI anchor of prostasomal CD59 and the effect of heat treatment on the prostasomes were also studied. Anti‐CD59 antibodies were used to block the protective effect of prostasomes on erythrocytes.

RESULTS: Both RE and PNH erythrocytes showed diminished complement‐mediated hemolysis after incubation with prostasomes. This was because of the transfer of CD59 from prostasomes to the red blood cells during pre‐incubation as evidenced by the hemolytic assay and flow‐cytometry. The efficacy of the prostasomes was affected by heat treatment and was totally lost at 100°C. Phosphatidylinositol phospholipase C broke the GPI anchor and released CD59 from prostasomes and the RE surface (after pre‐incubation with prostasomes) but not from the human PNH erythrocytes.

CONCLUSIONS: A transfer mechanism of CD59 takes place during pre‐incubation from prostasomes to erythrocytes lacking CD59 which supports the idea that transfer of prostasomal CD59 can protect cells from lysis elicited by C5b‐9. This might be a mechanism by which autologous and allogeneic cells are protected against complement attack in the genital tracts.
Restricted Entry of IgG into Male and Female Rabbit Reproductive Ducts Following Immunization with Recombinant Rabbit PH‐20
Authors: POMERING M. , JONES RUSSELL C. , HOLLAND MICHAEL K. , BLAKE ANITA E. , BEAGLEY KENNETH W.
Published: March 2003

PROBLEM: Successful immunocontraception using sperm antigens is dependent on achieving sufficient sperm‐specific antibody in the reproductive ducts to prevent fertilization. The blood : luminal barrier of the male and female reproductive ducts must be overcome for this to occur. We have, therefore, investigated the relative titers of antigen‐specific immunoglobulin G (IgG) in luminal fluids collected from male and female rabbit reproductive ducts following immunization with recombinant rabbit PH‐20 (rPH‐20).

METHOD OF STUDY: Male and female rabbits were immunized subcutaneously with rPH‐20 in Freund's adjuvant. Reproductive tract fluids and plasma were collected and assayed for specific IgG by enzyme‐linked immunoabsorbent assay (ELISA). Plasma anti‐rPH‐20 antibodies were tested for their ability to inhibit in vitro fertilization.

RESULTS: Plasma rPH‐20‐specific IgG titers of >21 × 106 were induced in bucks. Antibody levels in the rete testis and cauda epididymidis fluids were only 0.026 and 0.168% of plasma levels, respectively. Plasma IgG titers were >30 × 106 in does, but antibody levels in free flow vaginal fluid, free flow uterine fluid and free flow oviduct fluid were only 0.016, 0.078 and 0.072% of plasma levels, respectively. Induction of ovulation by administration of human chorionic gonadotrophin (hCG) significantly increased rPH‐20‐specific IgG only in free flow vaginal fluids. Plasma antibody from immunized rabbits inhibited in vitro fertilization but conception rates following mating of rPH‐20 immunized male to untreated female rabbits were not affected.

CONCLUSIONS: The IgG antibody entry into the reproductive ducts of both male and female rabbits is restricted to less than 0.2% of levels induced in plasma following subcutaneous immunization. This finding raises doubts about the suitability of rPH‐20 to induce immunocontraception in rabbits using strategies optimized for induction of a serum antibody response. Whether mucosal immunization strategies can achieve this remains to be tested.
Effect of treatment delay upon pulp and periodontal healing of traumatic dental injuries – a review article
Authors: Andreasen J.O. , Andreasen F.M. , Skeie A. , Hjørting‐Hansen E. , Schwartz O.
Published: March 2003

Abstract – Based on an analysis of the literature concerning parameters influencing the prognosis of traumatic dental injuries, few studies were found to have examined possible relationships between treatment delay and pulpal and periodontal ligament healing complications. It has been commonly accepted that all injuries should be treated on an emergency basis, for the comfort of the patient and also to reduce wound healing complications. For practical and especially economic reasons, various approaches can be selected to fulfill such a demand, such as acute treatment (i.e. within a few hours), subacute (i.e. within the first 24 h), and delayed (i.e. after the first 24 h). In this survey the consequences of treatment delay on pulpal and periodontal healing have been analyzed for the various dental trauma groups. Applying such a treatment approach to the various types of injuries, the following treatment guidelines can be recommended, based on our present rather limited knowledge of the effect of treatment delay upon wound healing. Crown and crown/root fractures: Subacute or delayed approach. Root fractures: Acute or subacute approach. Alveolar fractures: Acute approach (evidence however questionable). Concussion and subluxation: Subacute approach. Extrusion and lateral luxation: Acute or subacute approach (evidence however questionable). Intrusion: Subacute approach (evidence however questionable). Avulsion: If the tooth is not replanted at the time of injury, acute approach; otherwise subacute. Primary tooth injury: Subacute approach, unless the primary tooth is displaced into the follicle of the permanent tooth or occlusal problems are present; in the latter instances, an acute approach should be chosen. These treatment guidelines are based on very limited evidence from the literature and should be revised as soon as more evidence about the effect of treatment delay becomes available.
Augmentation of the narrow traumatized anterior alveolar ridge to facilitate dental implant placement
Authors: Oikarinen Kyösti S. , Sàndor George K. B. , Kainulainen Vesa T. , Salonen‐Kemppi Maarit
Published: March 2003

Abstract  – Traumatic tooth loss leads to alveolar resorption especially in sagittal direction. This can be due to avulsion of bone substance during the accident itself or due to resorption of the alveolar crest that takes place afterwards. Shortage of bone can prevent proper positioning of dental implants unless the volume of bone is increased before implantation. In the maxillary anterior area, this is also an esthetic problem. Several treatment modalities have been presented to augment the bone. This report reviews the latest literature on bone grafting, bone substitutes, guided bone regeneration, osteocompression and distraction which are potentially useful in the anterior maxilla. A special emphasis is paid to the versatility of using a crestal split osteotomy, by means of chisels and osteotomes to widen the narrow ridge. Three examples are illustrated showing onlay grafting, preservation of alveolar width with alloplastic coral material and lateral widening of a narrow maxillary alveolar ridge, using the crestal splitting technique.
Evidence for general cognitive ability (g) in heterogeneous stock mice and an analysis of potential confounds
Authors: Galsworthy M. J. , Paya‐Cano J. L. , Monleón S. , Plomin R.
Published: March 2003

The heterogeneous stock (HS) is a genetically outbred line of mice established more than 30 years ago from an 8‐way cross of C57BL/6, BALB/c, RIII, AKR, DBA/2, I, A/J and C3H inbred mouse strains. The present study compared the performance of 40 HS mice across a battery of diverse cognitive tasks under a variety of motivations. Indices of emotionality were also included in order to assess their influence on performance. All measures of ability loaded positively on an unrotated first principal component that accounted for 31% of the variance, suggesting the presence of a common factor of general cognitive ability (g) underlying all tasks. A first factor derived from anxiety indices correlated nonsignificantly with all cognitive tasks and nonsignificantly with this g factor, supporting the hypothesis that the factor is cognitive rather than temperamental in nature. The factor was also robust in relation to outliers and sex differences, accounting for 28% of the variance after removal of outlier individuals and also after correcting for variance owing to sex differences. A general cognitive ability (g) appears to underlie the performance of HS mice on a battery tapping diverse cognitive demands.
Multiple Cross Mapping (MCM) markedly improves the localization of a QTL for ethanol‐induced activation
Authors: Hitzemann R. , Malmanger B. , Cooper S. , Coulombe S. , Reed C. , Demarest K. , Koyner J. , Cipp L. , Flint J. , Talbot C. , Rademacher B. , Buck K. , McCaughran J.
Published: March 2003

This study examines the use of multiple cross mapping (MCM) to reduce the interval for an ethanol response QTL on mouse chromosome 1. The phenotype is the acute locomotor response to a 1.5‐g/kg i.p. dose of ethanol. The MCM panel consisted of the six unique intercrosses that can be obtained from the C57BL/6J (B6), DBA/2J (D2), BALB/cJ (C) and LP/J (LP) inbred mouse strains (N ≥ 600/cross). Ethanol response QTL were detected only with the B6xD2 and B6xC intercrosses. For both crosses, the D2 and C alleles were dominant and decreased ethanol response. The QTL information was used to develop an algorithm for sorting and editing the chromosome 1 Mit microsatellite marker set (http://www.jax.org). This process yielded a cluster of markers between 82 and 85 cM (MGI). Evidence that the QTL was localized in or near this interval was obtained by the analysis of a sample (n = 550) of advanced cross heterogenous stock animals. In addition, it was observed that one of the BXD recombinant inbred strains (BXD‐32) had a recombination in the interval of interest which produced the expected change in behavior. Overall, the data obtained suggest that the information available within existing genetic maps coupled with MCM data can be used to reduce the QTL interval. In addition, the MCM data set can be used to interrogate gene expression data to estimate which polymorphisms within the interval of interest are relevant to the QTL.
Recombinant factor VIIa in the management of surgery and acute bleeding episodes in children with haemophilia and high responding inhibitors
Authors: O'Connell Niamh , Mc Mahon Corrina , Smith Jude , Khair Kate , Hann Ian , Liesner Ri , Smith Owen P.
Published: March 2003

Summary.  The management of acute and surgical bleeding episodes in children with severe factor VIII or IX deficiency who develop high responding inhibitors presents a major therapeutic challenge to clinicians. Recombinant factor VIIa (rVIIa) is an effective, reliable and safe treatment that can be used to treat acute bleeding episodes prior to commencing an immune tolerance programme and to cover surgical procedures until the immune tolerance programme is successful. In a significant minority of patients, immune tolerance therapy is ineffective and an alternative haemostatic agent such as rVIIa is required for life‐long treatment. The present study evaluated the use of rVIIa in a paediatric setting. Twelve children, aged 1–16 years, were treated successfully with rVIIa to prevent surgical bleeding in 20 surgical procedures (19 central venous access device insertion or removal, 1 dental extraction). Minor postoperative haematomata developed in 2 out of 20 cases after regular rVIIa therapy had been discontinued and resolved with a short course of rVIIa in both cases. Three children had six life‐ or limb‐threatening bleeding episodes. All bleeding episodes resolved with regular rVIIa treatment although topical fibrin glue was needed in one child with a frenulum tear. One patient required two red cell transfusions for symptomatic anaemia resulting from two separate bleeding episodes. The rVIIa therapy was well tolerated and there was no evidence of treatment‐related complications. We conclude that rVIIa is the treatment of choice for the management of surgery and acute life‐ or limb‐threatening bleeding in children with haemophilia and high responding inhibitors.
Lack of multimer organization of von Willebrand factor in an acquired von Willebrand syndrome
Authors: Casonato A. , Pontara E. , Doria A. , Bertomoro A. , Cattini M. G. , Gambari P. F. , Girolami A.
Published: March 2003

Summary. We report a case of acquired von Willebrand syndrome (AVWS) in a 20‐year‐old‐woman with systemic lupus erythematosus, in whom severe bleeding complications followed kidney biopsy. Coagulation studies demonstrated undetectable levels of ristocetin‐induced platelet aggregation (RIPA), von Willebrand factor antigen (VWF:Ag) and VWF ristocetin cofactor activity (VWF:RCo), associated with significantly prolonged bleeding time; unlike type 3 von Willebrand disease (VWD), platelet VWF was reduced but not undetectable. The plasma VWF multimer pattern was characterized by the presence of only two bands, one of low molecular weight (MW) running as the protomer of plasma VWF in normals, the other of abnormally high MW without detectable intermediate multimers; this pattern resembles that of VWF present in endothelial cells. A search for an anti‐VWF antibody demonstrated the presence of an inhibitor at high titre. This anti‐VWF antibody did not interfere in the interaction of VWF with platelet glycoprotein (GP) Ib through the A1 domain, and did not react with the A2 domain of VWF; instead, it seemed to modify the relative representation of high and low MW VWF multimers released by normal human umbilical vein endothelial cells (HUVEC). After Azathioprine and corticosteroid treatment, the anti‐VWF antibody disappeared and the patient's haemostatic profile normalized, except for the platelet VWF content which still remained decreased. We suggest that the anti‐VWF antibody present in the AVWS described compromised both circulating VWF levels and their multimeric organization, inducing the maintenance of the multimer structure that VWF normally has before or in the early phase after secretion from endothelial cells.
Effect of laparoscopy on immune function
Authors: Gupta A , Watson D I
Published: March 2003

Background

Laparoscopic surgery is believed to lessen surgical trauma and so cause less disturbance of immune function. This may contribute to the rapid recovery noted after many laparoscopic operations. Preservation of both systemic and intraperitoneal immunity is particularly important in surgery for sepsis or cancer and so an understanding of the impact of laparoscopy on immune function is relevant.

Methods

Literature on immunological changes following laparoscopy and open surgery was identified from Medline, along with cross-referencing from the reference lists of major articles on the subject.

Results and discussion

Despite a few contradictory reports, systemic immunity appears to be better preserved after laparoscopic surgery than after open surgery. However, the local intraperitoneal immune system behaves in a particular way when exposed to carbon dioxide pneumoperitoneum; suppression of intraperitoneal cell-mediated immunity has been demonstrated in a number of studies. This feature may be clinically important and should be acknowledged when considering laparoscopic surgery in patients with malignancy or sepsis.
Acute compartment syndromes
Authors: Tiwari A , Haq A I , Myint F , Hamilton G
Published: March 2003

Background

Acute compartment syndrome is both a limb- and life-threatening emergency that requires prompt treatment. To avoid a delay in diagnosis requires vigilance and, if necessary, intracompartmental pressure measurement. This review encompasses both limb and abdominal compartment syndrome, including aetiology, diagnosis, treatment and outcome.

Methods

A Pubmed and Cochrane database search was performed. Other articles were cross-referenced.

Results and conclusion

Diagnosis of limb compartment syndrome is based on clinical vigilance and repeated examination. Many techniques exist for tissue pressure measurement but they are indicated only in doubtful cases, the unconscious or obtunded patient, and children. However, monitoring of pressure has no harmful effect and may allow early fasciotomy, although the intracompartmental pressure threshold for such an undertaking is still unclear. Abdominal compartment syndrome requires measurement of intra-abdominal pressure because clinical diagnosis is difficult. Treatment is by abdominal decompression and secondary closure. Both types of compartment syndrome require prompt treatment to avoid significant sequelae.
Structural investigation on the lipooligosaccharide fraction of psychrophilic Pseudoalteromonas haloplanktis TAC 125 bacterium
Authors: Corsaro M. Michela , Lanzetta Rosa , Parrilli Ermenegilda , Parrilli Michelangelo , Tutino M. Luisa
Published: March 2003

The core structure of the cell‐wall lipooligosaccharide (LOS) fraction of an Antarctic Gram‐negative bacterium, Pseudoalteromonas haloplanktis TAC 125 strain, was determined to be deacetylated alditols. These were obtained from native LOS fraction by O‐deacylation, dephosphorylation, reduction and finally N‐deacylation. Two novel structures were detected, the more highly represented molecule consisting of the following hexasaccharide chain: α‐d‐ManpNH2‐(1→3)‐β‐d‐Galp‐(1→4)‐α‐l‐glycero‐d‐manno‐Hepp‐(1→5)‐α‐d‐Kdo‐(2→6)‐β‐d‐GlcpNH2‐(1→6)‐d‐GlcNH2(ol) while the corresponding pentasaccharide, lacking the ManpNH2 residue, was less abundant.

To the best of our knowledge, the structural investigation presented here, mainly performed by NMR and MS methods, is the first report of the lipopolysaccharide fraction of a psychrophilic bacterium.
Signalling events underlying platelet aggregation induced by the glycoprotein VI agonist convulxin
Authors: Atkinson Ben T. , Stafford Margaret J. , Pears Catherine J. , Watson Steve P.
Published: March 2003

We have investigated the role of secretion and intracellular signalling events in aggregation induced by the glycoprotein (GP)VI‐selective snake venom toxin convulxin and by collagen. We demonstrate that aggregation induced by threshold concentrations of convulxin undergoes synergy with ADP acting via the P2Y12 receptor whereas there is no synergy via the P2Y1 receptor or with thromboxanes. On the other hand, apyrase, the P2Y12 receptor antagonist, AR‐C67085, and indomethacin only marginally inhibit aggregation induced by convulxin. In comparison, these inhibitors severely attenuate the response to collagen. In order to investigate whether the weak inhibitory action against convulxin is due to release of agonists other than ADP from dense granules, experiments were performed on murine platelets deficient in this organelle (pearl mice platelets). A slightly greater reduction in aggregation induced by convulxin was observed in pearl platelets than in the presence of inhibitors of ADP, but a maximal response was still attained. Importantly, inhibition of protein kinase C further reduced the response to convulxin in pearl platelets demonstrating a direct role for the kinase in aggregation. Chelation of intracellular Ca2+ with 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N,N′,N′‐tetraacetic acid (acetoxymethyl)ester (BAPTA‐AM) abolished aggregation induced by convulxin under all conditions. Activation of phospholipase C by convulxin was potentiated by ADP acting through the P2Y12 receptor. In conclusion, we show that Ca2+ and protein kinase C, but not release of the secondary agonists ADP and thromboxane A2, are required for full aggregation induced by convulxin, whereas the response induced by collagen shows a much greater dependence on secretion of secondary agonists.
Structural analysis of the lipopolysaccharide from the nontypable Haemophilus influenzae strain SB 33
Authors: Cox Andrew D. , Masoud Hussein , Thibault Pierre , Brisson Jean‐Robert , van der Zwan Mark , Perry Malcolm B. , Richards James C.
Published: March 2003

The structure of the core region of the lipopolysaccharide (LPS) from the nontypable Haemophilus influenzae strain SB 33 was elucidated. The LPS was subjected to a variety of degradative procedures. The structures of the derived oligosaccharide products were established by monosaccharide and methylation analyses, NMR spectroscopy and mass spectrometry. These analyses revealed a series of related phosphocholine (PCho) containing structures differing in the number of hexose residues. The results pointed to each species containing a conserved phosphoethanolamine (PEtn) substituted heptose‐containing trisaccharide inner‐core moiety. The major LPS glycoforms were identified as 2‐Hex, 3‐Hex and 4‐Hex species according to the number of hexose residues present.
The role of protein phosphorylation in human health and disease.
Authors: Cohen Philip
Published: March 2003

The reversible phosphorylation of proteins regulates almost all aspects of cell life, while abnormal phosphorylation is a cause or consequence of many diseases. Mutations in particular protein kinases and phosphatases gives rise to a number of disorders and many naturally occurring toxins and pathogens exert their effects by altering the phosphorylation states of intracellular proteins. In this lecture, I present an overview of the progress that is being made in developing specific inhibitors of protein kinases for the treatment of cancer and chronic inflammatory diseases and describe how recent advances in our understanding of the specificity and regulation of one particular protein kinase (GSK3) may facilitate the development of drugs to treat diabetes that would not have the potential to be oncogenic. I also discuss the exploitation of specific protein kinase inhibitors for the study of cell signalling and make recommendations for their effective use in cell‐based assays.
Physiological and biochemical characteristics of poly γ‐glutamate synthetase complex of Bacillus subtilis
Authors: Ashiuchi Makoto , Nawa Chizuko , Kamei Tohru , Song Jae‐Jun , Hong Seung‐Pyo , Sung Moon‐Hee , Soda Kenji , Yagi Toshiharu , Misono Haruo
Published: March 2003

An enzymatic system for poly γ‐glutamate (PGA) synthesis in Bacillus subtilis, the PgsBCA system, was investigated. The gene‐disruption experiment showed that the enzymatic system was the sole machinery of PGA synthesis in B. subtilis. We succeeded in achieving the enzymatic synthesis of elongated PGAs with the cell membrane of the Escherichia coli clone producing PgsBCA in the presence of ATP and d‐glutamate. The enzyme preparation solubilized from the membrane with 8 mm Chaps catalyzed ADP‐forming ATP hydrolysis only in the presence of glutamate; the d‐enantiomer was the best cosubstrate, followed by the l‐enantiomer. Each component of the system, PgsB, PgsC, and PgsA, was translated in vitro and the glutamate‐dependent ATPase reaction was kinetically analyzed. The PGA synthetase complex, PgsBCA, was suggested to be an atypical amide ligase.
Analysis of a putative voltage‐gated prokaryotic potassium channel
Authors: Ungar D. , Barth A. , Haase W. , Kaunzinger A. , Lewitzki E. , Ruiz T. , Reiländer H. , Michel H.
Published: March 2003

Most of the completely sequenced prokaryotic genomes contain genes of potassium channel homologues, but there is still not much known about the role of these proteins in prokaryotes. Here we describe the large‐scale overproduction and purification of a prokaryotic voltage‐gated potassium channel homologue, Kch, from Escherichia coli. After successful overproduction of the protein, a specific increase in the potassium permeability of the cells was found. Kch could be purified in large amounts using classical purification methods to prevent aggregation of the protein. The physiological state of the protein was revealed to be a homotetramer and the protein was shown to be localized to the cytoplasmic membrane of the cells. In the course of the localization studies, we found a specific increase in the density of the cytoplasmic membrane on Kch production. This was linked to the observed increase in the protein to lipid ratio in the membranes. Another observed change in the membrane composition was an increase in the cardiolipin to phosphatidylglycerol ratio, which may indicate a specific cardiolipin requirement of Kch. On the basis of some of our results, we discuss a function for Kch in the maintenance of the membrane potential in E. coli.
Phosphorylation of eukaryotic initiation factor 4E (eIF4E) at Ser209 is not required for protein synthesis in vitro and in vivo
Authors: McKendrick Linda , Morley Simon J. , Pain Virginia M. , Jagus Rosemary , Joshi Bhavesh
Published: March 2003

Eukaryotic translation initiation factor 4E (eIF4E) is essential for efficient translation of the vast majority of capped cellular mRNAs; it binds the 5′‐methylated guanosine cap of mRNA and serves as a nucleation point for the assembly of the 48S preinitiation complex. eIF4E is phosphorylated in vivo at residue 209 of the human sequence. The phosphorylated form is often regarded as the active state of the protein, with ribosome‐associated eIF4E enriched for the phosphorylated form and increased phosphorylation often correlated with upregulation of rates of protein synthesis. However, the only reported measured effect attributable to phosphorylation at the physiological site has been a relatively small increase in the affinity of eIF4E for the mRNA m7GTP cap structure. Here, we provide data to suggest that phosphorylation of eIF4E at Ser209 is not required for translation. eIF4E that is modified such that it cannot be phosphorylated (Ser209→Ala), is unimpaired in its ability to restore translation to an eIF4E‐dependent in vitro translation system. In addition, both the wild‐type and mutant forms of eIF4E interact equally well with eIF4G, with the phosphorylation of eIF4E not required to effect the change in conformation of eIF4G that is required for efficient cleavage of eIF4G by L‐protease. Furthermore, we show that wild‐type and phosphorylation‐site variants of eIF4E protein are equally able to rescue the lethal phenotype of eIF4E deletion in S. cerevisiae.
Solution structure of Pyrobaculum aerophilum DsrC, an archaeal homologue of the gamma subunit of dissimilatory sulfite reductase
Authors: Cort John R. , Mariappan S. V. Santhana , Kim Chang‐Yub , Park Min S. , Peat Thomas S. , Waldo Geoffrey S. , Terwilliger Thomas C. , Kennedy Michael A.
Published: March 2003

The solution structure of DsrC, an archaeal homologue of the γ subunit of dissimilatory sulfite reductase, has been determined by NMR spectroscopy. This 12.7‐kDa protein from the hyperthermophilic archaeon Pyrobaculum aerophilum adopts a novel fold consisting of an orthogonal helical bundle with a β hairpin along one side. A portion of the structure resembles the helix‐turn‐helix DNA‐binding motif common in transcriptional regulator proteins. The protein contains two disulfide bonds but remains folded following reduction of the disulfides. DsrC proteins from organisms other than Pyrobaculum species do not contain these disulfide bonds. A conserved cysteine next to the C‐terminus, which is not involved in the disulfide bonds, is located on a seven‐residue C‐terminal arm that is not part of the globular protein and is likely to dynamically sample more than one conformation.
Purification, characterization, and cDNA cloning of a novel soluble saxitoxin and tetrodotoxin binding protein from plasma of the puffer fish, Fugu pardalis
Authors: Yotsu‐Yamashita Mari , Sugimoto Atsuko , Terakawa Takahiro , Shoji Yuki , Miyazawa Teruo , Yasumoto Takeshi
Published: March 2003

Some species of puffer fish have been reported to possess both of tetrodotoxin and saxitoxin, which share one binding site on sodium channels. We purified a novel soluble glycoprotein that binds to these toxins from plasma of the puffer fish, Fugu pardalis, and named puffer fish saxitoxin and tetrodotoxin binding protein (PSTBP). PSTBP possessed a binding capacity of 10.6 ± 0.97 nmol·mg−1 protein and a Kd of 14.6 ± 0.33 nm for [3H]saxitoxin in equilibrium binding assays. [3H]Saxitoxin (10 nm) binding to PSTBPs was half‐inhibited by the presence of tetrodotoxin and saxitoxin at 12 µm and 8.5 nm, respectively. From the results of gel filtration chromatography (200 kDa) and SDS/PAGE (104 kDa), PSTBP was suggested to consist of noncovalently linked dimers of a single subunit. PSTBP was completely deglycosylated by glycopeptidase F, producing a single band at 42 kDa. Two highly homologous cDNAs to each other coding PSTBP (PSTBP1 and PSTBP2, the predicted amino‐acid identity 93%), were obtained from a cDNA library of F. pardalis liver. These proteins consisted to two tandemly repeated homologous domains. The predicted amino‐acid sequences of PSTBP1 and 2 were not homologous to that of saxiphilin, a reported saxitoxin binding protein, or sodium channels, but their N‐terminus sequences were homologous to that of the reported tetrodotoxin binding protein from plasma of Fugu niphobles, which has not been fully characterized. The partially homologous cDNA sequences to PSTBP1 and 2 were also found in expressed sequence tag clones of nontoxic flounders liver. Presumably, PSTBP is involved in accumulation and/or excretion of toxins in puffer fish.
The second ADF/cofilin actin‐binding site exists in F‐actin, the cofilin–G‐actin complex, but not in G‐actin
Authors: Blondin Laurence , Sapountzi Vasilia , Maciver Sutherland K. , Renoult Celine , Benyamin Yves , Roustan Claude
Published: March 2003

ADF/cofilins are actin binding proteins that bind actin close to both the N‐ and C‐termini (site 1), and we have found a second cofilin binding site (site 2) centered around helix 112–125 [Renoult, C., Ternent, D., Maciver, S.K., Fattoum, A., Astier, C., Benyamin, Y. & Roustan, C. (1999) J. Biol. Chem.274, 28893–28899]. We proposed a model in which ADF/cofilin intercalated between subdomains 1 and 2 of two longitudinally associated actin monomers within the actin:cofilin cofilament, explaining the change in twist that ADF/cofilins induce in the filament [McGough, A. Pope, B., Chiu, W. & Weeds, A. (1998) J. Cell Biol.138, 771–781]. Here, we have determined the fuller extent of the cofilin footprint on site 1 of actin. Site 1 is primarily the G‐actin binding site. Experiments with both peptide mimetics and fluorescently labeled cofilin suggest that site 2 only becomes available for cofilin binding within the filament, possibly due to motion between subdomains 1 and 2 within an actin monomer. We have detected motion between subdomains 1 and 2 of G‐actin by FRET induced by cofilin, to reveal the second cofilin‐binding site. This motion may also explain how cofilins inhibit the nucleotide exchange of actin, and why the actin:cofilin complex is polymerizable without dissociation.
Lactate dehydrogenase has no control on lactate production but has a strong negative control on formate production in Lactococcus lactis
Authors: Andersen Heidi W. , Pedersen Martin B. , Hammer Karin , Jensen Peter R.
Published: March 2003

A series of mutant strains of Lactococcus lactis were constructed with lactate dehydrogenase (LDH) activities ranging from below 1% to 133% of the wild‐type activity level. The mutants with 59% to 133% of lactate dehydrogenase activity had growth rates similar to the wild‐type and showed a homolactic pattern of fermentation. Only after lactate dehydrogenase activity was reduced ninefold compared to the wild‐type was the growth rate significantly affected, and the ldh mutants started to produce mixed‐acid products (formate, acetate, and ethanol in addition to lactate). Flux control coefficients were determined and it was found that lactate dehydrogenase exerted virtually no control on the glycolytic flux at the wild‐type enzyme level and also not on the flux catalyzed by the enzyme itself, i.e. on the lactate production. As expected, the flux towards the mixed‐acid products was strongly enhanced in the strain deleted for lactate dehydrogenase. What is more surprising is that the enzyme had a strong negative control (

=−1.3) on the flux to formate at the wild‐type level of lactate dehydrogenase. Furthermore, we showed that L. lactis has limited excess of capacity of lactate dehydrogenase, only 70% more than needed to catalyze the lactate flux in the wild‐type cells.
A Raman optical activity study of rheomorphism in caseins, synucleins and tau
Authors: Syme Christopher D. , Blanch Ewan W. , Holt Carl , Jakes Ross , Goedert Michel , Hecht Lutz , Barron Laurence D.
Published: March 2003

The casein milk proteins and the brain proteins α‐synuclein and tau have been described as natively unfolded with random coil structures, which, in the case of α‐synuclein and tau, have a propensity to form the fibrils found in a number of neurodegenerative diseases. New insight into the structures of these proteins has been provided by a Raman optical activity study, supplemented with differential scanning calorimetry, of bovine β‐ and κ‐casein, recombinant human α‐, β‐ and γ‐synuclein, together with the A30P and A53T mutants of α‐synuclein associated with familial cases of Parkinson's disease, and recombinant human tau46 together with the tau46 P301L mutant associated with inherited frontotemporal dementia. The Raman optical activity spectra of all these proteins are very similar, being dominated by a strong positive band centred at ≈ 1318 cm−1 that may be due to the poly(l‐proline) II (PPII) helical conformation. There are no Raman optical activity bands characteristic of extended secondary structure, although some unassociated β strand may be present. Differential scanning calorimetry revealed no thermal transitions for these proteins in the range 15–110 °C, suggesting that the structures are loose and noncooperative. As it is extended, flexible, lacks intrachain hydrogen bonds and is hydrated in aqueous solution, PPII helix may impart a rheomorphic (flowing shape) character to the structure of these proteins that could be essential for their native function but which may, in the case of α‐synuclein and tau, result in a propensity for pathological fibril formation due to particular residue properties.
A β‐lysine adenylating enzyme and a β‐lysine binding protein involved in poly β‐lysine chain assembly in nourseothricin synthesis in Streptomyces noursei
Authors: Grammel Nicolas , Pankevych Kvitka , Demydchuk Julia , Lambrecht Klaus , Saluz Hans‐Peter ,   Ullrich Keller, Krügel Hans
Published: March 2003

Nourseothricins (syn. Streptothricins), a group of nucleoside peptides produced by several streptomycete strains, contain a poly β‐lysine chain of variable length attached in amide linkage to the amino sugar moiety gulosamine of the nucleoside portion. We show that the nourseothricin‐producing Streptomyces noursei contains an enzyme (NpsA) of an apparent Mr 56 000 that specifically activates β‐lysine by adenylation but does not bind to it as a thioester. Cloning and sequencing of npsA from S. noursei including its flanking DNA regions revealed that it is closely linked to the nourseothricin resistance gene nat1 and some other genes on the chromosome possibly involved in nourseothricin biosynthesis. The deduced amino‐acid sequence revealed that NpsA is a stand‐alone adenylation domain with similarity to the adenylation domains of nonribosomal peptide synthetases (NRPS). Further analysis revealed that S. noursei contains a β‐lysine binding enzyme (NpsB) of about Mr 64 100 which can be loaded by NpsA with β‐lysine as a thioester. Analysis of the deduced amino‐acid sequence from the gene (npsB) of NpsB showed that it consists of two domains. The N‐terminal domain of ≈ 100 amino‐acid residues has high similarity to PCP domains of NRPSs whereas the 450‐amino‐acid C‐terminal domain has a high similarity to epimerization (E)‐domains of NRPSs. Remarkably, in this E‐domain the conserved H‐H‐motif is changed to H‐Q, which suggests that either the domain is nonfunctional or has a specialized function. The presence of one single adenylating β‐lysine activating enzyme in nourseothricin‐producing streptomycete and a separate binding protein suggests an iteratively operating NRPS‐module catalyses synthesis of the poly β‐lysine chain.