bluuebunny/crossref_metadata_2025 huggingface dataset:
Reactivity with erythroid and non‐erythroid tissues of a murine monoclonal antibody to a synthetic peptide having amino acid sequence common to cytoplasmic domain of human glycophorins C and D
Authors: King M.‐J. , Holmes C. H. , Mushens R. E. , Mawby W. , Reid M. E. , Scott M. L.
Published: March 2008

Three synthetic peptides encompassing the entire cytoplasmic polypeptide sequence (amino acid residues 82‐128) of glycophorin C (GPC) and glycophorin D (GPD) were used to immunize mice for the production of monoclonal antibodies (MoAbs). Only the synthetic peptide (GPC‐peptidel) corresponding to C‐terminal residues 112‐128 elicited a MoAb (named BGRL‐100) which could react with native and denatured GPC and GPD. We characterized BGRL‐100 by inhibition using GPC‐peptide 1 and red cell sialoglycoproteins. The ability of BGRL‐100 to interact with native GPC and GPD was assessed by immunoprecipitation with normal red cells (RBCs), and with denatured GPC and GPD by Western blotting of both normal RBCs and RBCs carrying GPC variants. Immunohistochemical staining of human tissue sections was performed using both BGRL‐100 and a rat MoAb (named BRAC‐1), which is specific for an extracellular domain of GPC and GPD. Both antibodies showed strong staining of erythroid lineage haemopoietic cells in fetal liver, sinusoids of adult liver and RBCs in the blood vessels of all tissues tested. Neither antibody reacted with epithelia from a range of human tissues. However, both MoAbs stained neural tissue in a distinctive fibrillar pattern. This suggests the presence of an analogue of erythroid GPC in neural tissues.
Diamond‐Blackfan anaemia: three patterns of in vitro response to haemopoietic growth factors
Authors: McGuckin Colin P. , Ball Sarah E. , Gordon‐Smith Edward C.
Published: March 2008

Culture of bone marrow from patients with Diamond‐Blackfan anaemia (DBA) has previously shown a variable progenitor response to growth factor stimulation. An extensive standardized study has now been undertaken to investigate the presence of distinct sub‐groups in this disorder. In vitro response of bone marrow progenitors to recombinant human growth factors, including stem cell factor, was examined in 18 DBA patients and five normal donors, assessing BFU‐E, CFU‐GM and CFU‐GEMM development. In 16 of the DBA patients a synergistic response to combinations of growth factors was observed with optimal growth in cultures containing erythropoietin, interleukin‐3 and stem cell factor. Growth factor induced erythroid response formed three distinct groups, based on BFU‐E numbers: type I (mean age 4.87 years) showed > 70% normal erythroid response; type II (mean age 13.87 years) showed < 70% normal; and type III (mean age 15.29 years) < 5% normal. CFU‐GM response also followed the trigrouping. The results suggest more than one pathogenic mechanism for the erythroid failure in DBA, indicating DBA may be composed of more than one distinct disorder, and further suggest the defect in DBA may not be confined to the erythroid series.
Analysis of molecular breakpoint and m‐RNA transcripts in a prospective randomized trial of interferon in chronic myeloid leukaemia: no correlation with clinical features, cytogenetic response, duration of chronic phase, or survival
Authors: Shepherd Patricia , Suffolk Ruth , Halsey Jim , Allan Norman
Published: March 2008

Two hundred and nineteen cases of Ph+ve CML and 15 Ph‐ve, BCR+ve CML cases have been analysed to determine the breakpoint site and its relationship to clinical features, cytogenetic response, duration of chronic phase and survival. 119 cases have had RNA analysis performed to determine the type of BCR/ABL transcript and have also been analysed in a similar way. Presenting features at diagnosis including age, sex, white‐cell count and platelet count showed no significant difference for those with 5’and 3’breakpoints and those with either b2a2 or b3a2 BCR/ABL transcripts. However, in a subgroup of patients whose presenting white‐cell count was <100×p109/1, those with b3a2 transcript did have a significantly higher platelet count.

Analysis by Sokal risk grouping showed no difference for 5’or 3’breakpoints but a trend for lower stage among those with b2a2 transcripts. No correlation was found either for genomic breakpoint site or BCR/ABL RNA transcript in terms of duration of chronic phase or survival. When stratified by randomized therapy, either interferon‐α or standard chemotherapy, no difference was noted in relation to genomic breakpoint site or BCR/ABL transcript. Cytogenetic response was not related to the molecular findings.
The receptor for urokinase plasminogen activator is present in plasma from healthy donors and elevated in patients with paroxysmal nocturnal haemoglobinuria
Authors: Rønne Ebbe , Pappot Helle , Grøndahl‐Hansen Jan , Høyer‐Hansen Gunilla , Plesner Torben , Hansen Niels Ebbe , Danø Keld
Published: March 2008

The urokinase plasminogen activator (uPA) is a proteolytic enzyme which converts the proenzyme plasminogen to the active serine protease plasmin. A cell surface receptor for uPA (uPAR) is attached to the cell membrane by a glycosyl‐phosphatidylinositol anchor. Binding of uPA to uPAR leads to an enhanced plasmin formation and thereby an amplification of pericellular proteolysis. We have shown previously that uPAR is expressed on normal blood monocytes and granulocytes, but is deficient on affected blood monocytes and granulocytes in patients with paroxysmal nocturnal haemoglobinuria (PNH), and that uPAR is present in plasma from these patients. In this study a newly established sensitive enzyme‐linked immunosorbent assay (ELISA) has been applied for quantttation of uPAR in plasma. Unexpectedly, we found that uPAR is not only present in PNH plasma but also in plasma from healthy individuals. In 39 healthy individuals the mean plasma‐uPAR value ±SD was 31 ± 15 pm, median 28 (range 11‐108), and the corresponding value for six PNH patients was 116±67 pm, median 90 (range 61‐228). The elevated uPAR‐level in PNH patients was highly significant (Mann‐Whitney test; P < 0.0001), and may possibly contribute to the propensity for thrombosis in PNH by inhibition of the fibrinolytic system. Binding of pro‐uPA by uPAR in plasma may interfere with the appropriate binding of pro‐uPA to cell‐bound uPAR and therefore inhibit cell‐associated plasmin generation and fibrinolysis. It is likely that the uPAR in normal plasma reflects the overall level of activity of the uPAR‐mediated cell surface proteolysis. The present ELISA may be used for studies of uPAR levels in plasma from patients with conditions in which this activity might be increased, such as cancer and inflammatory disorders. Future studies will determine if uPAR in plasma is a parameter of clinical importance in these diseases.
Severe autoimmune protein S deficiency in a boy with idiopathic purpura fulminans
Authors: Bergmann F. , Hoyer P. F. , ANGELO S. Vigano' D' , Mazzola G. , Oestereich C. , Barthels M. , D'Angelo A.
Published: March 2008

Idiopathic purpura fulminans usually occurs in young children and is frequently preceeded by a preparatory viral or bacterial infection. Following a severe streptococcal pharyngitis, an 8‐year‐old boy developed purpura fulminans with disseminated intravascular coagulation and severe protein S deficiency (total antigen <0.05μ/ml). Despite generous plasma infusions, skin necrosis progressed rapidly into compartment syndrome which required fasciotomy and skin grafting and resulted in the loss of three digits of the right foot. Total protein S remained low for over a month despite plasma supplementation and complete normalization of protein C levels. A polyclonal anti‐protein S IgG was demonstrated in the patient's plasma, which decreased to 25% of baseline titre after 1 month and was undetectable 6 months after purpura fulminans, when plasma protein S had returned to normal. Transient, isolated and severe deficiencies of protein S have been reported in patients with idiopathic purpura fulminans and a previous preparatory infection. Autoimmune protein S deficiency may play a key role in the aetiopathogenesis of idiopathic purpura fulminans.
Expression of granulocyte colony‐stimulating factor receptor on CD10‐positive human B‐cell precursors
Authors: Inukai Takeshi , Sugita Kanji , Iijima Kiyomu , Tezuka Toru , Goi Kumiko , Kojika Satoru , Shiraishi Kyoko , Kagami Keiko , Nakazawa Shinpei
Published: March 2008

We examined the expression of CD10 and G‐CSF receptor (G‐CSFR) on the lymphoid population of mononuclear cells obtained from bone marrow (BM) using two‐colour analysis. In the BM of children with ALL in remission, the CD10+ population was significantly increased (20.6 ± 5.1% compared with that of controls (2.5 ± 0.5%). More than half (61.3 ± 2.9%) of the CD10+ cells coexpressed G‐CSFR, but not CD13. These results indicate G‐CSFR+ B‐cell precursors are markedly increased in BM of ALL in remission, suggesting the probable involvement of G‐CSF in the human early B‐cell ontogeny.
Detection of monoclonal plasma cells in the peripheral blood stem cell harvests of patients with multiple myeloma
Authors: Witzig Thomas E. , Gertz Morie A. , Pineda Alvaro A. , Kyle Robert A. , Greipp Philip R.
Published: March 2008

We evaluated the harvest product from 47 patients undergoing peripheral blood (BP) stem cell collections for monoclonal plasma cells (PC) using a sensitive immunofluorescence technique. 60% (28/47) had docu mented tumour cells in the apheresis product. The 32 patients in plateau had a mean of 1.62 x 106 PC/1 v 74.64 x 106 in 15 relapsed patients (P < 0.01). 32% (6/19) of patients without any tumour cells in the initial sample had them detected on a subsequent apheresis sample. In four cases (all treated with GM‐CSF) small numbers of tumour cells were detected initially but became undetectable on a subsequent sample.
Application of a polymorphic Y microsatellite to the detection of post bone marrow transplantation chimaerism
Authors: Lo Y.‐M. D. , Noakes L. , Roux E. , Jeannet M. , Chapuis B. , Fleming K. A. , Wainscoat J. S.
Published: March 2008

A highly sensitive Y‐microsatellite amplification system was developed for the detection of post bone marrow transplantation (BMT) chimaerism. The system is able to detect the DNA equivalent of a single male cell in a background of 105 female cells. For clinical applications it has the distinct advantage that the polymorphic nature of the PCR product allows the checking of the genuineness of a positive PCR signal by reference to its allelic type. This extra degree of precision enhances the accuracy of Y‐PCR for clinical diagnosis.
Identification of biclonal (duplex) leukaemic cells expressing either CD4+/CD8‐ or CD4‐/CD8+ from a patient with adult T‐cell leukaemia/lymphoma
Authors: Kondo Seiji , Kotani Tomio , Tsumori Shinichiro , Narahara Shinichiro , Aratake Yatsuki , Kobayashi Masako , Takahashi Masayoshi , Inoue Shouhei , Ohtaki Sachiya
Published: March 2008

A 24‐year‐old Japanese woman was admitted to our hospital in 1987 with a chief complaint of skin eruptions, and was diagnosed as having chronic ATLL. In 1993 the leucocyte count increased gradually to 126.0x109/l with 91.5% abnormal lymphocytes expressing two different types of antigenicity, either CD4+/CD8‐ or CD4‐/CD8+. Monoclonal integration of human T‐cell lymphotropic virus type‐I proviral DNA was detected at different sites of the genomic DNA in each cell type. These studies clearly indicate that CD4+/CD8‐ and CD4‐/CD8‐ leukaemic cells originated from two independent clones.
Biological control methods of pest insects in oilseed rape
Authors: Hokkanen H. M. T.
Published: March 2008

Parasitoids, predators, insect pathogenic fungi, nematodes and microsporidia have all shown promise in the biological suppression of the key target pest, the pollen beetle Meligethes aeneus in rapeseed, as well as in the control of other rapeseed pests. Under favourable conditions pollen beetles are locally and temporarily already under efficient natural control by one or several of these agents, but overall the pollen beetle remains the number one pest of rapeseed in Finland, as well as elsewhere in Europe. Conservation biological control to enhance natural control appears the most feasible approach to solve the problem. Field and simulation data are used to demonstrate how a simple change in the tillage practice may be enough to obtain efficient control of the pollen beetle. Several other options are, or will be available, subject to successful registration of microbial products based on the fungi Metarhizium anisopliae and Beauveria bassiana, and the nematode Steinernema feltiae. Strategies and methods of applying these agents have been developed to facilitate either inundative treatments, or ecosystem restoration to (re‐)establish the presence of these agents in agricultural fields. Critical components of agricultural practice have been identified for maximising control by biological agents within a functioning IPM system.
Different methods of monitoring susceptibility of oilseed rape beetles to insecticides
Authors: Thieme T. , Drbal U. , Gloyna K. , Hoffmann U.
Published: March 2008

The resistance of oilseed rape pollen beetles to lambda‐cyhalothrin has increased in Germany over the period 2002 to 2007. Oilseed rape pollen beetles are very good at dispersing, therefore field results should always be compared with those of laboratory experiments. The response of oilseed rape pollen beetles to active substances may not be the same as that to formulated products. Bioassays in which beetles are exposed to insecticides on either complete inflorescences or flower buds give different results. It is evident that samples of the genus Meligethes collected in the field can include several different species. Monitoring for insecticide resistance should be done using only the true oilseed rape pollen beetle.
Habitat heterogeneity overrides the species–area relationship
Authors: Báldi András
Published: March 2008

Aim  The most obvious, although not exclusive, explanation for the increase of species richness with increasing sample area (the species–area relationship) is that species richness is ultimately linked to area‐based increases in habitat heterogeneity. The aim of this paper is to examine the relative importance of area and habitat heterogeneity in determining species richness in nature reserves. Specifically, the work tests the hypothesis that species–area relationships are not positive if habitat heterogeneity does not increase with area.

Location  Sixteen nature reserves (area range 89–11,030 ha) in central Hungary.

Methods  Four‐year faunistic inventories were conducted in the reserves involving c. 70 fieldworkers and 65 taxonomists. CORINE 50,000 land‐cover maps were used for calculating the heterogeneity of the reserve landscape (number of habitat types, number of habitat patches and total length of edges).

Results  Large reserves were less heterogeneous than small reserves, probably because large reserves were established in large blocks of unproductive land whereas small reserves tended to be in more fertile land. In total, 3975 arthropod species were included in the analysis. The slope of the species–area relationship was positive only for Neuroptera and Trichoptera. There was no significant relationship in the other nine taxa examined (Collembola, Acari, Orthoptera, Thysanoptera, Coleoptera, Araneae, Diplopoda, Chilopoda, Diptera). The density (number of species ha−1) of all species, however, showed a positive correlation with heterogeneity.

Main conclusions  The general lack of fit of species–area relationships in this study is inconsistent with most previous published studies. Importantly, and unlike many other studies, habitat heterogeneity was not correlated with reserve area in the studied system. In the absence of this source of covariation, stronger relationships were identified that suggested a fundamental link between species richness and habitat heterogeneity. The results indicate that habitat heterogeneity rather than area per se is the most important predictor of species richness in the studied system.
The association between cheese consumption and cardiovascular risk factors among adults
Authors: Houston D. K. , Driver K. E. , Bush A. J. , Kritchevsky S. B.
Published: March 2008

Background  Although current dietary guidelines recommend limiting foods high in fat and saturated fat, such as high‐fat dairy, the effect of cheese consumption on body composition and cardiovascular risk factors is largely unknown.

Methods  Participants from a US population‐based survey, NHANES III, aged 25–75 years who completed a food frequency questionnaire and had measures of body composition and cardiovascular risk factors were included (n = 10 872). Linear regression was used to compare anthropometrics, blood lipids, blood pressure and blood glucose across categories of cheese consumption (combined full and low‐fat).

Results  In women, more frequent cheese consumption was associated with higher HDL‐C and lower LDL‐C (p for trend, < 0.05). However, in men, more frequent cheese consumption was associated with a higher body mass index (BMI), waist circumference, HDL‐C and LDL‐C, and diastolic blood pressure (p for trend, < 0.05). Men consuming 30 +  servings/month had significantly higher BMI, waist circumference, and diastolic blood pressure compared to nonconsumers (P < 0.05).

Conclusions  More frequent cheese consumption was associated with less favourable body composition and cardiovascular risk profile in men, but with a more favourable cardiovascular risk profile in women. However, the type of cheese consumed by men and women may have differed resulting in opposing trends on body composition and cardiovascular risk factors.
Assessing dietary patterns in Barbados highlights the need for nutritional intervention to reduce risk of chronic disease
Authors: Sharma S. , Cao X. , Harris R. , Hennis A. J. M. , Wu S.‐Y. , Leske M. C. ,   for the Barbados National Cancer Study Group
Published: March 2008

Background  The dietary habits of the Caribbean have been changing to include more fast foods and a less nutrient dense diet. The aims of this study are to examine dietary patterns in Barbados and highlight foods for a nutritional intervention.

Methods  Four‐day food diaries collected from control participants in the population‐based, case–control Barbados National Cancer Study (BNCS).

Results  Forty‐nine adult participants (91% response) completed the diaries providing 191 days of dietary data. Total energy intake was almost identical to data collected 5‐years earlier in the Barbados Food Consumption and Anthropometric Survey 2000, but the percent energy derived from fat was from 2.1% to 5.2% higher. Sugar intake exceeded the Caribbean recommendation almost four‐fold, while intakes of calcium, iron (women only), zinc and dietary fibre were below recommendations. Fish and chicken dishes were the two largest sources of energy and fat. Sweetened drinks and juices provided over 40% of total sugar intake.

Conclusions  These data provide existing dietary patterns and strongly justify a nutritional intervention program to reduce dietary risk factors for chronic disease. The intervention could focus on the specific foods highlighted, both regarding frequency and amount of consumption. Effectiveness can be evaluated pre‐ and post‐intervention using our Food Frequency Questionnaire developed for BNCS.
Nutrition support in major burn injury: case analysis of dietetic activity, resource use and cost implications
Authors: Windle E. M.
Published: March 2008

Background  Patients with major burn injury are among the most catabolic and nutritionally vulnerable of all hospital admissions. A considerable amount of dietetic intervention is often required. To date, UK practice has not been described in detail. This study aimed to identify nutritional interventions, resource use and associated costs for a typical patient admitted with major burns.

Methods  A 28‐year‐old male with 43.5% total body surface area flame injury was selected for study. Dietetic, medical and pharmacy records were reviewed for data regarding nutritional status, interventions and product use. Costs of dietetic staffing, nutrition support products, related ancillary items and hospital food provision were calculated.

Results  The patient required 68 days of nutrition support, including 40 days of nasogastric tube feeding. Seventy per cent of bedside reviews resulted in dietetic intervention. Initiation of nutrition support, substrate use, and frequency of biochemical and weight monitoring were largely in compliance with practice guidelines. Overall cost of nutritional care for the inpatient episode was £1377.

Conclusion  Work is now required to assess current nutrition practices across different UK centres and for a range of burn severities, to establish a baseline from which resource and financial requirements can ultimately be developed.
Three persons with multiple disabilities accessing environmental stimuli and asking for social contact through microswitch and VOCA technology
Authors: Lancioni G. E. , O'Reilly M. F. , Singh N. N. , Sigafoos J. , Oliva D. , Severini L.
Published: March 2008

Background  Direct access to environmental stimuli and opportunity to ask for social contact/attention may be considered highly relevant objectives for persons with multiple disabilities. We assessed the possibility of enabling three of these persons (two children and one adolescent) to combine two microswitches (for accessing environmental stimuli) and a Voice Output Communication Aid (VOCA), which allowed them to ask for caregiver's attention.

Methods  Initially, the participants were required to use each of the two microswitches individually and then together. Thereafter, they were taught to use the VOCA. Eventually, the VOCA was available together with the microswitches, and the participants could use any of the three.

Results  The results, which support preliminary data on this topic, showed that all participants (1) were able to operate the two microswitches as well as the VOCA; and (2) used all three of them consistently when they were simultaneously available.

Conclusions  Teaching persons with multiple disabilities to combine a VOCA with conventional microswitches may enrich their general input, emphasize their active social role and eventually enhance their social image.
Behavioural characteristics associated with dementia assessment referrals in adults with Down syndrome
Authors: Adams D. , Oliver C. , Kalsy S. , Peters S. , Broquard M. , Basra T. , Konstandinidi E. , McQuillan S.
Published: March 2008

Background  Behavioural changes associated with dementia in Down syndrome are well documented, yet little is known about the effect of such behaviours on carers and referral. By comparing the behavioural and cognitive profiles of individuals referred for a dementia assessment with those of individuals not referred, some insight can be gained into behavioural characteristics that initiate referral for specialist support or interventions.

Method  Forty‐six adults with Down syndrome were divided into two groups dependent upon method of entry into the study; post‐referral to a specialist service for older adults with intellectual disabilities and Down syndrome for a dementia assessment (n = 17) or after receiving information sent out to day centres and residential homes (n = 29). These groups were compared on established measures of dementia alongside two informant measures of behaviour.

Results  Those referred for a dementia assessment evidenced scores indicative of cognitive decline on both informant and direct Neuropsychological Assessments and showed more behavioural excesses, but not deficits, and lower socialisation and coping skills than those in the comparison group. Carers of those referred for a dementia assessment reported a greater impact of behavioural excesses on staff than on the individual showing the behaviour in contrast to the comparison group.

Conclusion  The behavioural differences between those referred and the comparison group suggest that two factors are involved in the instigation of a referral for a dementia assessment: the nature of the behavioural presentation (excesses rather than deficits) and the effect of that behavioural change upon the care staff.
Nitric oxide synthesis and signalling in plants
Authors: WILSON IAN D. , NEILL STEVEN J. , HANCOCK JOHN T.
Published: March 2008

As with all organisms, plants must respond to a plethora of external environmental cues. Individual plant cells must also perceive and respond to a wide range of internal signals. It is now well‐accepted that nitric oxide (NO) is a component of the repertoire of signals that a plant uses to both thrive and survive. Recent experimental data have shown, or at least implicated, the involvement of NO in reproductive processes, control of development and in the regulation of physiological responses such as stomatal closure. However, although studies concerning NO synthesis and signalling in animals are well‐advanced, in plants there are still fundamental questions concerning how NO is produced and used that need to be answered. For example, there is a range of potential NO‐generating enzymes in plants, but no obvious plant nitric oxide synthase (NOS) homolog has yet been identified. Some studies have shown the importance of NOS‐like enzymes in mediating NO responses in plants, while other studies suggest that the enzyme nitrate reductase (NR) is more important. Still, more published work suggests the involvement of completely different enzymes in plant NO synthesis. Similarly, it is not always clear how NO mediates its responses. Although it appears that in plants, as in animals, NO can lead to an increase in the signal cGMP which leads to altered ion channel activity and gene expression, it is not understood how this actually occurs.

NO is a relatively reactive compound, and it is not always easy to study. Furthermore, its biological activity needs to be considered in conjunction with that of other compounds such as reactive oxygen species (ROS) which can have a profound effect on both its accumulation and function. In this paper, we will review the present understanding of how NO is produced in plants, how it is removed when its signal is no longer required and how it may be both perceived and acted upon.
Subgingival microbiota of shallow periodontal pockets in individuals after head and neck irradiation
Authors: Leung W. K. , Jin L. J. , Samaranayake L. P. , Chlu G. K. C.
Published: March 2008

This study aimed at investigating the subgingival plaque microorganisms of shallow pockets (≤5 mm) in subjects who previously received irradiation in the head and neck region for treatment of nasopharyngeal carcinoma. Direct microscopy and anaerobic culture were used. Subgingival paper point samples were taken from 6 tooth‐sites (one/sextant) per subject for direct microscopy (n= 108). Another set of paper points was taken from the deepest of the previously selected sites (one per subject) with: group A) no bleeding on probing to the sulcus depth (n= 9) and group B) bleeding on probing to the sulcus depth (n= 6) for microscopic and anaerobic culture study. Under the microscope, the microflora was found to be a complex mixture comprising gram‐positive and gram‐negative cocci, rods and filaments, fusiforms, curved rods and spirochetes. Low level of fungi were observed and mycelia were occasionally detected. There was no significant variation in the plaque bacterial morphotypes observable according to sites of isolation and no significant difference between group A and group B in morphotypes of the different microflora. The predominant cultivable microflora comprised several species of facultative and obligate anaerobic bacteria: Gemella, Peptostreptococcus, Staphylococus, Stomatococcus, Streptococcus, Actinomyces, Eubacterium, Lactobacillus, Propionibacterium, Neisseria, Veillonella, Bacteroides, Campylobacter, Capnocytophaga, Fusobacterium, Kingella, Porphyromonas and Prevotella species. There was no difference between the two groups except the significantly higher proportion of Kingella dentrificans isolated from group B sites. However, colonization of the gingival sulcus in these individuals by microbes that are normal flora of: skin (Peptostreptococcus prevotii and Propionibacterium granulosum) and gut (Eubacterium aerofaciens, Fusobacterium mortiferum and Fusobacterium varium) was detected. These findings appear to suggest that the major components of the subgingival microflora of shallow sites in previously head‐and neck‐irradiated individuals are similar to that of gingivitis sites in the normal population although they may contain bacterial or fungal species uncommon in normal subjects.
Identification of bacterial species on or in crevicular epithelial cells from healthy and periodontally diseased patients using DNA‐DNA hybridization
Authors: Dibart S. , Skobe Z. , Snapp K. R. , Socransky S. S. , Smith C. M. , Kent R.
Published: March 2008

The purpose of this investigation was to identify bacterial species present on or in crevicular epithelial cells in healthy and diseased sites using DNA probes. In order to achieve this aim, further improvements were made in the separation of unattached bacteria from those adherent to epithelial cells isolated from the human gingival crevice or periodontal pocket. Then the DNA probes were used to determine the prevalence of detectable DNA from 15 microbial species on or in crevicular epithelial cells. One sample was taken from a single subgingival site in each of 51 individuals ranging in age from 19 to 45 years. Samples were taken from 27 sites of clinically healthy subjects and 24 samples were taken from subjects having periodontally diseased sites. DNA‐DNA hybridization indicated that a majority of epithelial cells from healthy sites (63%) were in contact with or harbored Streptococcus oralis. On the other hand, species such as Bacteroides forsythus, Prevotella intermedia, Capnocytophaga ochracea and Campylobacter rectus were more frequently detected in elevated numbers in periodontally diseased sites. Cluster analysis of the microbial profiles generally aggregated subjects with and without periodontitis into separate cluster groups. The cluster patterns suggest the possibility that microbial complexes will be, in part, determined by the receptors available on the epithelial cells.
Cross‐reactivity of specific antibodies directed to heat shock proteins from periodontopathogenic bacteria of human origin
Authors: Hinode D. , Nakamura R. , Grenier D. , Mayrand D.
Published: March 2008

This study describes the immunological characterization of two different classes of heat shock proteins isolated from periodontopathogenic bacteria. Analysis of the JV‐terminal amino acid sequence of a 74‐kDa protein from Bacteroides forsylhus showed a high degree of homology with the DnaK protein from Escherichia coli. However, this heat shock protein from B. forsythus reacted very weakly with a commercial anti‐DnaK polyclonal antibody by dot‐blotting. GroEL‐like proteins isolated from Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and B. forsythus showed a high degree of homology of their ^‐terminal amino acid sequences. In general, polyclonal antibodies raised against each GroEL‐like protein showed a high level of cross‐reactivity. The cross‐reactivity of antibodies to bacterial DnaK‐like proteins was much more limited. Our findings suggest that DnaK‐and GroEL‐like proteins from periodontal pathogens are well conserved and that the GroEL‐like proteins resemble each other more closely.
Development of gene expression system in a marine diatom using viral promoters of a wide variety of origin
Authors: Sakaue Kunihiro , Harada Hisashi , Matsuda Yusuke
Published: March 2008

Promoter sequences of the cytomegalovirus (PCMV), the rous sarcoma virus long terminal repeat (PRSV‐LTR) and the cauliflower mosaic virus 35s (PCaMV35s) were ligated with the β‐glucuronidase (GUS) gene, uidA, and were introduced into cells of the marine diatom, Phaeodactylum tricornutum. Transformants were selected on a 100 mg l−1 Zeocin plate, and Zeocin‐resistant clones were further selected by the occurrence of GUS activity. Two to 10 GUS‐positive clones were obtained, and GUS activities in these transformants did not change in response to changes in ambient CO2 concentration except that the PRSV‐LTR was weakly activated in air. These results indicate that a wide spectrum of viral promoters originating from mammalian, avian and plant hosts can operate as constitutive promoters in a marine diatom. The CO2 responsive promoter sequence of the chloroplastic carbonic anhydrase gene in P. tricornutum (Pptca1) with a deleted initiator region was ligated with the minimal region of the PCMV followed by uidA and was introduced into P. tricornutum. GUS expression in the resulting transformants was clearly regulated by CO2, that is, GUS expression was stimulated in air to about 10‐fold than that in cells grown in 5% CO2. However, the CO2 response disappeared when the core regulatory region of Pptca1 (−76 to −11 bp) was removed. The regulative function of the endogenous diatom promoter was thus maintained after fusion with an extrinsic viral promoter. These results indicate that diatom cells accommodate a wide range of transcriptional system from beyond the plant kingdom and that an efficient transcriptional system could potentially be constructed in marine diatoms by selecting an appropriate set of viral promoter and functional cis elements.
Transfusion support for a patient with McLeod phenotype without chronic granulomatous disease and with antibodies to Kx and Km
Authors: Bansal I. , Jeon H.‐R. , Hui S. R. , Calhoun B. W. , Manning D. W. , Kelly T. J. , Lee S. , Baron B. W.
Published: March 2008

Background and Objectives  Kell antigens are encoded by the KEL gene on the long arm of chromosome 7. Kx antigen is encoded by the XK gene on the short arm of the X chromosome. Kell and Kx proteins in the red cell membrane are covalently linked by a disulphide bond. The McLeod phenotype is characterized by weakened expression of antigens in the Kell blood group system, absence of Km and Kx antigens, and acanthocytosis. It has an X‐linked mode of inheritance with transmission through carrier females. Some males with the McLeod syndrome also have chronic granulomatous disease (CGD). It is generally believed that patients with non‐CGD McLeod may develop anti‐Km but not anti‐Kx, but that those with CGD McLeod can develop both anti‐Km and anti‐Kx.

Materials and Methods  We present serological data, DNA genotyping and gene sequencing, monocyte monolayer assay and neutrophil oxidative burst test from a patient with the McLeod phenotype without clinical evidence of CGD.

Results  We report here the second example of a patient with non‐CGD McLeod who developed anti‐Kx in addition to anti‐Km. Sequencing of our patient's XK gene confirmed the presence of a mutation resulting in a premature stop codon and lack of Kx protein in the red cell membrane, which is consistent with the diagnosis of McLeod syndrome. Neutrophil oxidative burst test was normal, indicating that our patient did not have CGD. The challenge of providing 10 compatible blood units for multiple surgeries was met.

Conclusion  The second case of a rare entity, a patient with non‐CGD McLeod who developed anti‐Kx and anti‐Km, was managed successfully with a combination of autologous donations and procurement of compatible units from national and international sources.
Efficacy and tolerability of human fibrinogen concentrate administration to patients with acquired fibrinogen deficiency and active or in high‐risk severe bleeding
Authors: Danés A. Farriols , Cuenca L. Gallur , Bueno S. Rodríguez , Mendarte Barrenechea L. , Ronsano J. Bruno Montoro
Published: March 2008

Background and Objectives  Fibrinogen deficiency is a cause for massive haemorrhage whose management in emergency situations is the subject of debate. Plasma‐derived fibrinogen concentrates are indicated for reversing the haemorrhagic diathesis found in congenital and acquired deficiencies.

Materials and Methods  We report on the results of an observational study that evaluated the effects of fibrinogen concentrates in patients suffering from various forms of acquired severe hypofibrinogenaemia with life‐threatening consumptive thrombo‐haemorrhagic disorders (surgery, trauma and digestive haemorrhage), or underlying disease states that limit fibrinogen synthesis (hepatic dysfunction, haematological malignancies).

Results  Sixty‐nine patients were identified and included, in whom most of the processes (62%) corresponded to consumptive hypofibrinogenaemia. After a median dose of 4 g, a mean absolute increase of 1·09 g/l in plasma fibrinogen was measured and coagulation parameters were significantly improved (P < 0·001). Mortality rates of 32·3% and 44·2% were reported after 24 h and 72 h, respectively.

Conclusion  We conclude that the administration of fibrinogen concentrates in unresponsive, life‐threatening haemorrhage with acquired hypofibrinogenaemia improves laboratory measures of coagulation, and may also be life saving. Although observational in nature, our data indicate a direct relationship between plasma fibrinogen levels and survival in acquired fibrinogen deficiency. Further studies are warranted to ascertain a clear relationship between fibrinogen levels and survival.
Mitochondrial mechanisms of death responses in pancreatitis
Authors: Odinokova Irina V , Sung Kai‐Feng , Mareninova Olga A , Hermann Kip , Gukovsky Ilya , Gukovskaya Anna S
Published: March 2008

Pancreatitis is a severe and frequently lethal disorder, a major cause of which is alcohol abuse. Parenchymal cell death is a major complication of pancreatitis. In experimental models of acute pancreatitis, acinar cells have been shown to die through both necrosis and apoptosis, the two principal pathways of cell death. The severity of experimental acute pancreatitis correlates directly with the extent of necrosis and inversely with apoptosis. Thus, understanding the regulation of apoptosis and necrosis is becoming exceedingly important in investigations of the pathogenesis and treatment of pancreatitis. Over the past decade, the mitochondria have emerged as a master regulator of cell death in various physiological and pathological processes. Release of mitochondrial cytochrome c into the cytosol is a central event in apoptosis, whereas mitochondrial depolarization resulting in ATP depletion leads to necrosis. The present review focuses on the mitochondrial mechanisms of death responses in pancreatitis, with emphasis on mitochondrial membrane permeabilization and its role in the balance between apoptosis and necrosis in acute pancreatitis, and alcohol's effects on death responses of pancreatitis.
Impaired receptor‐mediated endocytosis: Its role in alcohol‐induced apoptosis
Authors: Casey Carol A , Lee Serene ML , Aziz‐Seible Razia , McVicker Benita L
Published: March 2008

Hepatocyte apoptosis, inflammation, and fibrosis are prominent features of liver disease in general and of alcoholic liver injury in particular. Although the link between these processes remains unclear, one universal characteristic of liver injury is the induction of hepatocellular damage, which results in the generation of apoptotic bodies. Work from our laboratory over the last several years has studied the effect of ethanol administration on the process of apoptosis and a role for altered endocytosis in alcoholic apoptosis. We initially focused our research on the hepatocyte by examining endocytosis using the asialoglycoprotein receptor (ASGP‐R) pathway as a model and we identified multiple ethanol‐induced impairments in receptor function. We also showed that uptake of apoptotic bodies is impaired in hepatocytes isolated from ethanol‐fed animals compared to controls, and that this impairment is linked to altered ASGP‐R function. Recent work from our laboratory is examining a link between ethanol‐impaired ASGP‐R function, apoptotic body accumulation, and inflammation in the liver. We are particularly interested in data showing that factors produced by Kupffer cells incubated with apoptotic bodies can lead to production of tumor necrosis factor‐alpha and interleukin‐6, and that this effect is exacerbated in the setting of alcohol administration. In addition, we have preliminary data showing that media from Kupffer cell cultures incubated with apoptotic bodies can induce hepatocyte killing. The goal of our future work is to show that inadequate removal of apoptotic cells, in part via altered receptor‐mediated endocytosis, plays a role in the course of pathogenesis of alcoholic liver injury.
Oxidative stress in alcoholic liver disease: Role of NADPH oxidase complex
Authors: De Minicis Samuele , Brenner David A
Published: March 2008

Alcohol is a well‐known risk factor for liver damage and is one of the major causes of liver disease worldwide. Chronic intake of alcohol, over a certain limit, inevitably leads to hepatic steatosis. If the injury persists, steatosis with concomitant tumor necrosis factor‐α and other cytokines, progresses to steatohepatitis, fibrosis and finally cirrhosis. Among the multiple factors involved in the process of alcohol‐induced liver injury, a crucial role is played by oxidative stress. Several mechanisms during ethanol metabolism result in reactive oxygen species (ROS) production. Although the main site of ethanol metabolism is hepatocytes, other mechanisms are involved in alcohol‐induced liver injury. Specifically, in the ROS production activity, an important role is played by the NADPH oxidase complex. NADPH oxidase is expressed in hepatocytes, hepatic stellate cells and Kupffer cells in the liver. Studying NADPH oxidase gives new insights into alcohol‐induced liver damage and provides new direction for future therapeutic strategies.
Association analysis among polymorphisms of the various genes and chronic alcoholic pancreatitis
Authors: Maruyama Katsuya , Harada Shoji , Yokoyama Akira , Naruse Satoru , Hirota Masahiko , Nishimori Isao , Otsuki Makoto
Published: March 2008

Chronic and excessive consumption of alcohol is an important factor responsible for the onset of pancreatitis. However, the incidence of chronic pancreatitis in heavy drinkers differs in individuals, suggesting that these individual differences may involve various genetic and environmental factors. In the present study, we investigated an association of alcoholic pancreatitis with polymorphisms of the various genes related to metabolism of the oxidative compounds. We analyzed polymorphisms of NADPH‐quinone oxidoreductase 2 (NQO2), multidrug resistance 1 (MDR1), alcohol dehydrogenase 1B (ADH1B) and lipoprotein lipase (LPL). The subjects consisted of 53 patients with chronic alcoholic pancreatitis (AlCP), 54 alcoholic patients without pancreatic dysfunction (Alc), and 42 healthy individuals. DNA samples were prepared from the peripheral blood of all subjects, and the genetic mutations were analyzed by polymerase chain reaction and restriction fragment length polymorphism methods. The ADH1B gene frequencies were significantly different between healthy controls and Alc patients (P < 0.001), and also between AlCP and Alc patients (P < 0.05). However, no significant difference was found between healthy controls and AlCP patients. The gene frequencies of MDR1 (3435C > T) and MDR1 (2677G > A/T) of patients with AlCP or Alc were different when compared with healthy controls, although the difference was not significant. The NQO2 and LPL genes showed no relation with Alc and AlCP patients. The ADH1B*1 gene frequency in AlCP was significantly lower compared with Alc. We speculate that the ADH1B*1 gene may function by reducing vulnerability to the onset of alcoholic pancreatitis. Other genes analyzed in the present study lacked association with AlCP.
Molecular basis for the synergy between alcohol and hepatitis C virus in hepatocarcinogenesis
Authors: Koike Kazuhiko , Tsutsumi Takeya , Miyoshi Hideyuki , Shinzawa Seiko , Shintani Yoshizumi , Fujie Hajime , Yotsuyanagi Hiroshi , Moriya Kyoji
Published: March 2008

Overwhelming lines of epidemiological evidence have indicated that persistent infection with hepatitis C virus (HCV) is a major risk for the development of hepatocellular carcinoma (HCC). In addition, heavy alcohol use has been linked with earlier progression to HCC in chronic hepatitis C patients. However, in the pathogenesis of HCV‐associated HCC, it still remains controversial as to whether the virus plays a direct or an indirect role, and as to how alcohol operates in the acceleration of HCC development. Several studies using transgenic mouse models, in which the core protein of HCV has an oncogenic potential, indicate that HCV is directly involved in hepatocarcinogenesis, although other factors such as continuous inflammation or environmental factors seem also to play a role. The downstream events of the HCV core protein expression in the transgenic mouse HCC model are segregated into two pathways. One is the augmented production of oxidative stress in the absence of inflammation along with the attenuation of some scavenging systems in the putative preneoplastic stage with steatosis in the liver. The other pathway is the alteration in cellular gene expression and intracellular signaling, including the mitogen‐activated protein kinase cascade. The combination of these pathways would explain the unusually high incidence and multicentric nature of HCC development in HCV infection. In addition, alcohol feeding in this animal model further activated the two pathways synergistically with HCV, leading to an earlier development of HCC. Such a synergy would reveal the molecular basis for the acceleration of HCC development by alcohol in HCV infection.
Alcohol, cofactors and the genetics of hepatocellular carcinoma
Authors: Yu Mimi C , Yuan Jian‐Min , Lu Shelly C
Published: March 2008

Heavy alcohol consumption, chronic infection with the hepatitis B virus (HBV) or the hepatitis C virus (HCV), tobacco smoking, and diabetes are risk factors for hepatocellular carcinoma (HCC). In the Los Angeles Non‐Asian HCC Study, heavy alcohol intake was shown to exhibit synergistic effects with viral hepatitis (HBV, HCV) and diabetes in the causation of HCC among individuals with joint exposures. Although chronic infection with HBV is recognized as the most important causal factor for HCC in humans, only a minority of HBV carriers eventually develop HCC, suggesting the presence of important cofactors in HBV‐related HCC. In the Guangxi/China HCC Study, a 20‐fold difference in HCC risk was observed between individuals possessing the least versus the most favorable cytokine genotypes for hepatitis B viral clearance. Experimental studies have indicated an important role for one‐carbon metabolism in HCC development. In both the Los Angeles and Guangxi studies, low‐activity genotypes (reduced enzymatic activities) of methylenetetrahydrofolate reductase (MTHFR) and high‐activity genotypes (enhanced enzymatic activities) of thymidylate synthase (TYMS), both of which discourage the misincorporation of uracil into DNA, were shown to be associated with a reduced risk for HCC.