bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of Sprouted and Midge‐Damaged Wheat Kernels Using Near‐Infrared Hyperspectral Imaging
Authors: Singh Chandra B. , Jayas Digvir S. , Paliwal Jitendra , White Noel D. G.
Published: April 2009

Sprout damage which results in poor breadmaking quality due to enzymatic activity of α‐amylase is one of the important grading factors of wheat in Canada. Potential of near‐infrared (NIR) hyperspectral imaging was investigated to detect sprouting of wheat kernels. Artificially sprouted, midge‐damaged, and healthy wheat kernels were scanned using NIR hyperspectral imaging system in the range of 1000–1600 nm at 60 evenly distributed wavelengths. Multivariate image analysis (MVI) technique based on principal components analysis (PCA) was applied to reduce the dimensionality of the hyperspectral data. Three wavelengths 1101.7, 1132.2, and 1305.1 nm were identified as significant and used in analysis. Statistical discriminant classifiers (linear, quadratic, and Mahalanobis) were used to classify sprouted, midge‐damaged, and healthy wheat kernels. The discriminant classifiers gave maximum accuracy of 98.3 and 100% for classifying healthy and damaged kernels, respectively.
Functional Properties of 7s Globulin Extracted from Cowpea Vicilins
Authors: Zhang Mei‐Li , Gao Ju‐Lin , Yang Hai‐Xia
Published: April 2009

The seed of cowpea (Vigna unguiculata L.) is rich in protein and the amino acid profiles of the meal are suitable for human dietary products, but little is known about the structure and chemical properties of the protein extracted from this legume. This study determined the functional properties of two selected cowpea cultivars and their solubility, emulsifying capacity, surface hydrophobicity, and thermal stability. Seeds of red and black cowpea were surface sterilized and the 7s globulin was isolated and purified using column chromatography with Sephacryl S‐300 (Hi‐Prep 26/60) gel column. Also, SDS‐PAGE and protein structure were analyzed using biochemical procedures. At high ionic strength (μ = 0.5), cowpea 7s globulin fraction exhibited better solubility for a wide range of pH levels, higher emulsifying capacities, and greater thermal stability than those obtained at low ionic strength (μ = 0.08). The lowest solubility was observed at pH 5.3–6.4 at the low ionic strength. Emulsifying capacities at high protein concentration were greater when compared with low protein concentration. Tm values of black cowpea globulin fraction were higher than those of red cowpea globulin fraction, whereas the surface hydrophobicity of the globulin fraction in red cowpea was larger than that in black cowpea.
Identification of non-fermenting Gram-negative bacteria of clinical importance by an oligonucleotide array
Authors: Su Siou Cing , Vaneechoutte Mario , Dijkshoorn Lenie , Wei Yu Fang , Chen Ya Lei , Chang Tsung Chain
Published: April 2009

Many species of non-fermenting Gram-negative bacilli (non-fermenters) are important opportunistic and nosocomial pathogens. Identification of most species of non-fermenters by phenotypic characteristics can be difficult. In this study, an oligonucleotide array was developed to identify 38 species of clinically relevant non-fermenters. The method consisted of PCR-based amplification of 16S–23S rRNA gene intergenic spacer (ITS) regions using bacterial universal primers, followed by hybridization of the digoxigenin-labelled PCR products with oligonucleotide probes immobilized on a nylon membrane. A total of 398 strains, comprising 276 target strains (i.e. strains belonging to the 38 species to be identified) and 122 non-target strains (i.e. strains not included in the array), were analysed by the array. Four target strains (three reference strains and one clinical isolate) produced discrepant identification by array hybridization. Three of the four discordant strains were found to be correctly identified by the array, as confirmed by sequencing of the ITS and 16S rRNA genes, with the remaining one being an unidentified species. The sensitivity and specificity of the array for identification of non-fermenters were 100 and 96.7 %, respectively. In summary, the oligonucleotide array described here offers a very reliable method for identification of clinically relevant non-fermenters, with results being available within one working day.
Comparison of different methods of determining β-lactam susceptibility in clinical strains of Pseudomonas aeruginosa
Authors: Torres Eva , Villanueva Rosa , Bou Germán
Published: April 2009

One hundred and one randomly selected (2003–2005) clinical isolates ofPseudomonas aeruginosawere used to assess the quantitative (MIC) and qualitative (susceptibility category) agreement between the microdilution broth reference method (RM) and disc diffusion (DD), Etest and the VITEK 2 automated susceptibility test system for determination of the susceptibility ofP. aeruginosato piperacillin (PIP), PIP–tazobactam (TZP), ceftazidime (CAZ), aztreonam (ATM) cefepime (FEP) and imipenem (IMP). The results obtained by the RM were compared with those obtained by the other methods. The RM and DD were performed according to CLSI criteria. Etest and VITEK 2 were according to the manufacturer's instructions. The Advanced Expert System (AES), which interprets MICs generated by VITEK 2, was modified with new rules of interpretation. Overall, VITEK 2 showed the lowest MIC90values for the six antibiotics. The RM categorical testing (susceptibility and resistance) rates withP. aeruginosawere 11.8 and 88.1 for PIP, 22.7 and 77.2 for TZP, 14.8 and 78.2 for CAZ, 12.8 and 54.4 for ATM, 16.8 and 75.3 for FEP, and 7.9 and 90.1 for IMP, respectively. Very major errors (false susceptible) were only detected for ATM and FEP with DD and for IMP with three methods. Major errors (false resistant) were generally acceptable for all antibiotics except TZP. VITEK 2 yielded a high level of minor errors (trends toward false susceptibility), mainly with CAZ and FEP. A good agreement was obtained for all antibiotics/methods assayed, thus highlighting the importance of the AES for categorization ofβ-lactam susceptibility inP. aeruginosa.
Commensal bacteria can enter colonic epithelial cells and induce proinflammatory cytokine secretion: a possible pathogenic mechanism of ulcerative colitis
Authors: Ohkusa Toshifumi , Yoshida Tsutomu , Sato Nobuhiro , Watanabe Sumio , Tajiri Hisao , Okayasu Isao
Published: April 2009

Interleukin 2 (IL-2)- and IL-10-knockout mice develop spontaneous colitis under conventional but not germ-free conditions, suggesting that commensal bacteria play an important role in the pathogenesis of colitis. However, interactions between commensal bacteria and colonic epithelial cells have not been fully investigated. We therefore assessed the ability of various commensal bacteria and probiotics to adhere to and invade colonic epithelial cells. Effects of the bacteria on production of proinflammatory cytokines were also measured. Commensal bacteria, including mucosal organisms isolated from ulcerative colitis (UC) patients, such asFusobacterium varium, reported as a possible pathogen in UC,Bacteroides vulgatus,Escherichia coliandClostridium clostridioforme, as well as their type strains and probiotics, were assessed for their ability to adhere to and invade colonic epithelial cells using two cell lines, SW-480 and HT-29. Our experiments employed co-incubation, a combination of scanning and transmission electron microscopy and recovery of bacteria from infected-cell lysates.F. variumand several other commensal bacteria, but not probiotics, adhered to colonic epithelial cells and invaded their cytoplasm. ELISA and real-time PCR revealed that the host cells, particularly those invaded byF. varium, showed significant increases in IL-8 and TNF-αconcentrations in supernatants, with elevation of IL-8, TNF-α, MCP-1 and IL-6 mRNAs. Furthermore, IL-8 and TNF-αexpression and nuclear phosphorylated NF-κB p65 expression could be immunohistochemically confirmed in inflamed epithelium with cryptitis or crypt abscess in UC patients. Certain commensal bacteria can invade colonic epithelial cells, activating early intracellular signalling systems to trigger host inflammatory reactions.
Efficient diagnosis of tuberculous meningitis by detection of Mycobacterium tuberculosis DNA in cerebrospinal fluid filtrates using PCR
Authors: Haldar Sagarika , Sharma Neera , Gupta V. K. , Tyagi Jaya Sivaswami
Published: April 2009

Tuberculous meningitis (TBM) is the most devastating form of meningitis and prompt diagnosis holds the key to its management. Conventional microbiology has limited utility and nucleic acid-based methods have not been widely accepted for various reasons. In view of the paucibacillary nature of cerebrospinal fluid (CSF) and the recent demonstration of free Mycobacterium tuberculosis DNA in clinical specimens, the present study was designed to evaluate the utility of CSF ‘filtrates’ for the diagnosis of TBM using PCR. One hundred and sixty-seven CSF samples were analysed from patients with ‘suspected’ TBM (n=81) and a control group including other cases of meningitis or neurological disorders (n=86). CSF ‘sediments’ and ‘filtrates’ were analysed individually for M. tuberculosis DNA by quantitative real-time PCR (qRT-PCR) and conventional PCR. Receiver-operating characteristic curves were generated from qRT-PCR data and cut-off values of 84 and 30 were selected for calling a ‘filtrate’ or ‘sediment’ sample positive, respectively. Based on these, TBM was diagnosed with 87.6 % and 53.1 % sensitivity (P <0.001) in ‘filtrates’ and ‘sediments’, respectively, and with 92 % specificity each. Conventional devR and IS6110 PCR were also significantly more sensitive in ‘filtrates’ versus ‘sediments’ (sensitivity of 87.6 % and 85.2 % vs 31 % and 39.5 %, respectively; P <0.001). The qRT-PCR test yielded a positive likelihood ratio of 11 and 6.6 by analysing ‘filtrate’ and ‘sediment’ fractions, respectively, which establishes the superiority of the ‘filtrate’-based assay over the ‘sediment’ assay. PCR findings were separately verified in 10 confirmed cases of TBM, where M. tuberculosis DNA was detected using devR PCR assays in ‘sediment’ and ‘filtrate’ fractions of all samples. From this study, we conclude that (i) CSF ‘filtrates’ contain a substantial amount of M. tuberculosis DNA and (ii) ‘filtrates’ and not ‘sediments’ are likely to reliably provide a PCR-based diagnosis in ‘suspected’ TBM patients.
Multiplex PCR reveals a high rate of nasopharyngeal pneumococcal 7-valent conjugate vaccine serotypes co-colonizing indigenous Warao children in Venezuela
Authors: Rivera-Olivero Ismar A. , Blommaart Martijn , Bogaert Debby , Hermans Peter W. M. , de Waard Jacobus H.
Published: April 2009

Knowledge of co-colonization with multiple pneumococcal serotypes is becoming very important in the light of both serotype replacement and switching as a result of vaccination. Co-colonization has been reported to occur in up to 30 % of carriers, especially in populations with highStreptococcus pneumoniaecarriage rates. For the determination of co-colonization, single colonies of nasopharyngeal specimens are serotyped with the Quellung method, a costly method with a low sensitivity. Here we explore the use of a multiplex PCR to identify simultaneous carriage of the capsular serotypes targeted by the 7-valent conjugate vaccine. We applied this multiplex PCR to 50 primary cultures from the nasopharyngeal swabs of healthy Warao Amerindian children, a population with a high pneumococcal carriage rate, most of them with vaccine serotypes, and we identified a second serotype in 20 % (n=10) of the pneumococci carriers. These results were confirmed by detailed serotyping of multiple colonies isolated from the primary culture with the Quellung method. We conclude that the multiplex PCR is a sensitive, simple and cost-effective method for detecting multiple serotypes in nasopharyngeal cultures, and thus might be useful for the monitoring of pneumococcal colonization over time, especially in the surveillance of nasopharyngeal colonization after conjugate vaccination.
Bartholin's abscess caused by hypermucoviscous Klebsiella pneumoniae
Authors: Pinsky Benjamin A. , Baron Ellen J. , Janda J. Michael , Banaei Niaz
Published: April 2009

Klebsiella pneumoniae serogroups displaying the hypermucoviscosity phenotype are associated with a distinct clinical syndrome characterized by liver abscesses, bacteraemia and metastatic lesions. We describe here what we believe to be the first reported case of hypermucoviscous K. pneumoniae causing a superficial Bartholin's abscess in the absence of systemic involvement.
Community-acquired meticillin-resistant Staphylococcus aureus in Palestine
Authors: Kaibni Maha H. , Farraj Mohammad A. , Adwan Kamel , Essawi Tamer A.
Published: April 2009

Community-acquired meticillin-resistantStaphylococcus aureus(CA-MRSA) is becoming an important public-health problem. This study attempted to investigateS. aureusand MRSA colonization in nasal swabs obtained from 843 patients without a history of hospitalization at the time of hospital admission and from 72 health-care workers chosen for comparison. Of the patients,S. aureuswas detected in 218/843 (25.9 %) and MRSA in 17/843 (2.0 %). Of the health-care workers,S. aureuswas detected in 15/72 (20.8 %) and MRSA in 10/72 (13.9 %). The majority of the 27 MRSA isolates exhibited a sensitivity pattern expected for CA-MRSA. Multilocus restriction fragment typing resolved the isolates into eight restriction fragment types. The predominant restriction fragment types were AAACCAA and AAAAAAA, accounting for 51.9 % (14/27) of the MRSA isolates and included CC5 and CC1 groups, respectively. This study thus demonstrated the transmission of CA-MRSA strain types into a health-care setting, emphasizing the need for implementation of a revised set of control measures in both hospital and community settings.
Two cases of failed ceftriaxone treatment in pharyngeal gonorrhoea verified by molecular microbiological methods
Authors: Tapsall John , Read Phillip , Carmody Christopher , Bourne Christopher , Ray Sanghamitra , Limnios Athena , Sloots Theo , Whiley David
Published: April 2009

Diagnostic, genotypic and antibiotic-resistance determinants ofNeisseria gonorrhoeaewere analysed by molecular methods to verify the failure of ceftriaxone treatment in two cases of pharyngeal gonorrhoea. Monoplex assays were needed to define competitive inhibition of a positiveChlamydiaPCR in a duplex assay. DifferentpenAchanges were detected in theN. gonorrhoeaeisolated from the two cases. These were associated with raised ceftriaxone MICs of 0.03 and 0.016 mg l−1, which may have contributed to the treatment failures in these cases.
Burkholderia mallei cellular interactions in a respiratory cell model
Authors: Whitlock Gregory C. , Valbuena Gustavo A. , Popov Vsevolod L. , Judy Barbara M. , Estes D. Mark , Torres Alfredo G.
Published: April 2009

Burkholderia mallei is a facultative intracellular pathogen that survives and replicates in phagocytic cell lines. The bacterial burden recovered from naïve BALB/c mice infected by intranasal delivery indicated that B. mallei persists in the lower respiratory system. To address whether B. mallei invades respiratory non-professional phagocytes, this study utilized A549 and LA-4 respiratory epithelial cells and demonstrated that B. mallei possesses the capacity to adhere poorly to, but not to invade, these cells. Furthermore, it was found that B. mallei was taken up by the murine alveolar macrophage cell line MH-S following serum coating, an attribute suggestive of complement- or Fc receptor-mediated uptake. Invasion/intracellular survival assays of B. mallei-infected MH-S cells demonstrated decreased intracellular survival, whilst a type III secretion system effector bopA mutant strain survived longer than the wild-type. Evaluation of the potential mechanism(s) responsible for efficient clearing of intracellular organisms demonstrated comparable levels of caspase-3 in both the wild-type and bopA mutant with characteristics consistent with apoptosis of infected MH-S cells. Furthermore, challenge of BALB/c mice with the bopA mutant by the intranasal route resulted in increased survival. Overall, these data suggest that B. mallei induces apoptotic cell death, whilst the BopA effector protein participates in intracellular survival.
Vertical Helicobacter pylori transmission from Mongolian gerbil mothers to pups
Authors: Oshio Ichiro , Osaki Takako , Hanawa Tomoko , Yonezawa Hideo , Zaman Cynthia , Kurata Satoshi , Kamiya Shigeru
Published: April 2009

To identify the time frame and route of mother-to-child Helicobacter pylori infection, a Mongolian gerbil model was used. Four-week-old female Mongolian gerbils were infected with H. pylori, and then mated with uninfected males 2 months after infection. The offspring were sacrificed weekly after birth, and then serum, mother's milk from the stomach and gastric tissues were obtained from pups. Anti-H. pylori antibody titres were measured in sera and maternal milk using an ELISA. The stomach was cut in two in the sagittal plane, and then H. pylori colonization in mucosa was confirmed by culture and real-time RT-PCR in one specimen and by immunochemical staining in the other. Faeces and oral swabs were obtained from infected mothers, and H. pylori 16S rRNA was measured using real-time RT-PCR. H. pylori was not identified in cultures from the gastric mucosa of pups delivered by infected mothers, but H. pylori 16S rRNA was detected from 4 weeks after birth, suggesting that Mongolian gerbil pups become infected via maternal H. pylori transmission from 4 weeks of age. The anti-H. pylori antibody titre in sera of pups from infected mothers was maximum at 3 weeks of age and then rapidly decreased from 4 weeks of age. High antibody titres in mother's milk were detected during the suckling period, and GlcNAcα was detectable at 2–4 weeks of age, but disappeared as the offspring aged. Thus H. pylori seems to infect Mongolian gerbil pups from 4 weeks of age, in parallel with decreasing GlcNAcα expression in the gastric mucosa. These results suggested that H. pylori infection of Mongolian gerbil pups occurs via faecal–oral transmission from an infected mother.
Glucocorticoid‐induced tumor necrosis factor receptor stimulation enhances the multifunctionality of adoptively transferred tumor antigen‐specific CD8+ T cells with tumor regression
Authors: Imai Naoko , Ikeda Hiroaki , Tawara Isao , Wang Linan , Wang Lijie , Nishikawa Hiroyoshi , Kato Takuma , Shiku Hiroshi
Published: April 2009

We have reported for the first time the significance of effector T‐cell multifunctionality in antitumor immunity, suggesting that the appearance of multifunctional/polyfunctional tumor‐specific CD8+ T cells in vivo is a critical determinant of the success of antitumor immunotherapy, and a strategy to induce multifunctionality in effector cells is required for the successful immunotherapy of hosts with progressing tumor. Glucocorticoid‐induced tumor necrosis factor receptor (GITR) stimulation has been shown to enhance antitumor immune response. However, its functional impact on adoptively transferred T cells remains unclear. Here, we analyzed the impact of GITR stimulation in vivo on the functional profiles of adoptively transferred CD8+ T cells specific for murine fibrosarcoma CMS5. GITR stimulation was found to enhance multifunctionality (interferon (IFN)‐γ and tumor necrosis factor (TNF)‐α production and CD107a mobilization as a degranulation marker) in transferred cells at the single‐cell level. These cells exhibited upregulated expression of CD25 in draining lymph nodes and increased infiltration in tumor. Mice that received T‐cell therapy with GITR stimulation showed reduced Foxp3+CD4+ T cells among tumor infiltrating lymphocytes and increased in vivo cytotoxic T lymphocytes (CTL) activity even with progressing tumor, resulting in enhanced tumor regression. These data strengthen the idea that effector T‐cell multifunctionality is a sensitive immune correlate for successful immunotherapy against malignancy and provide an immunological rationale for effective T‐cell therapy combined with GITR stimulation. (Cancer Sci 2009; 100: 1317–1325)
Cytoplasmic tethering is involved in synergistic inhibition of p53 by Mdmx and Mdm2
Authors: Ohtsubo Chihiro , Shiokawa Daisuke , Kodama Masami , Gaiddon Christian , Nakagama Hitoshi , Jochemsen Aart G. , Taya Yoichi , Okamoto Koji
Published: April 2009

The mdm2 and mdmx oncogenes play essential yet nonredundant roles in synergistic inactivatiosn of p53. However, the biochemical mechanism by which Mdmx synergizes with Mdm2 to inhibit p53 function remains obscure. Here we demonstrate that, using nonphosphorylatable mutants of Mdmx, the cooperative inhibition of p53 by Mdmx and Mdm2 was associated with cytoplasmic localization of p53, and with an increase of the interaction of Mdmx to p53 and Mdm2 in the cytoplasm. In addition, the Mdmx mutant cooperates with Mdm2 to induce ubiquitination of p53 at C‐terminal lysine residues, and the integrity of the C‐terminal lysines was partly required for the cooperative inhibition. The expression of subcellular localization mutants of Mdmx revealed that subcellular localization of Mdmx dictated p53 localization, and that cytoplasmic Mdmx tethered p53 in the cytoplasm and efficiently inhibited p53 activity. RNAi‐mediated inhibition of Mdmx or introduction of the nuclear localization mutant of Mdmx reduced cytoplasmic retention of p53 in neuroblastoma cells, in which cytoplasmic sequestration of p53 is involved in its inactivation. Our data indicate that cytoplasmic tethering of p53 mediated by Mdmx contributes to p53 inactivation in some types of cancer cells. (Cancer Sci 2009; 100: 1291–1299)
Malignant mesothelioma‐associated antigens recognized by tumor‐infiltrating B cells and the clinical significance of the antibody titers
Authors: Shigematsu Yoshiki , Hanagiri Takeshi , Kuroda Koji , Baba Tetsuro , Mizukami Makiko , Ichiki Yoshinobu , Yasuda Manabu , Takenoyama Mitsuhiro , Sugio Kenji , Yasumoto Kosei
Published: April 2009

Malignant pleural mesothelioma (MPM) is difficult to diagnose at an early stage. The present study attempted to obtain a tumor‐specific antibody against MPM derived from tumor‐infiltrating B lymphocytes in MPM by using a xenotransplanted severe combined immunodeficiency (SCID) mouse model, and to identify the antigens recognized by the antibodies. Among the antigen–antibody relationships, the clinical usefulness of antibody titers in the sera was evaluated from the viewpoint of diagnosis of MPM and monitoring of therapeutic effects. Tumor tissue specimens from two patients with MPM were engrafted subcutaneously in SCID mice and blood samples were obtained and pooled every 2 weeks after xenotransplantation until 14 weeks when the mice were killed. A cDNA library was constructed from the mRNA of a MPM cell line (K921MSO). Immunoscreening of the libraries was carried out by serological identification of antigens by a recombinant expression cloning method (SEREX) and four antigens were identified as MPM‐associated antigens. Among them, antibody titers against two antigens, Gene‐X and thrombospondin‐2 (THBS‐2), were analyzed by phage plaque assay as the first step. ELISA systems correlated with the phage plaque assay to detect antibody titers against the two antigens were constructed using 20‐mer peptides of the antigen‐coding genes. The cut‐off value was decided by the average and standard deviation of normal healthy persons. Antibody against Gene‐X was detected in 10 out of 18 (55.6%) mesothelioma patients and antibody against THBS‐2 was detected in 16 out of 18 (88.9%) mesothelioma patients. No patients with lung cancer regardless of asbestos exposure exhibited positive antibody titer against the two antigens. Furthermore, the serum antibody titers decreased after surgical treatment of MPM and increased after recurrence of the disease. The titers of the antibodies against Gene‐X and THBS‐2 could be used as tumor markers for the diagnosis and follow up of patients with MPM. (Cancer Sci 2009; 100: 1326–1334)
Comparison of the prevalence of fructose and lactose malabsorption across chronic intestinal disorders
Authors: BARRETT J. S. , IRVING P. M. , SHEPHERD S. J. , MUIR J. G. , GIBSON P. R.
Published: April 2009

Background  Fructose malabsorption, lactose malabsorption and an early rise in breath hydrogen after lactulose (ERBHAL) may play roles in induction of symptoms in gastrointestinal conditions.

Aim  To compare prevalence and interactions of fructose malabsorption, lactose malabsorption and ERBHAL among healthy subjects and those with chronic intestinal disorders using consistent breath hydrogen testing methodologies.

Methods  Consecutive series of Caucasian patients with Crohn’s disease (n = 91), ulcerative colitis (56), functional gastrointestinal disorders (FGID) (201), coeliac disease (136) and 71 healthy volunteers underwent breath hydrogen testing using lactulose, fructose and lactose.

Results  Early rise in breath hydrogen after lactulose occurred more commonly in healthy controls (39%) than in Crohn’s disease (20%) and untreated coeliac disease (14%; P < 0.008), but not FGID (27%), ulcerative colitis (26%) or treated coeliac disease (29%). Fructose malabsorption was more frequent in Crohn’s disease (61%) than other groups (33–44%, P < 0.05). Lactose malabsorption was most common in Crohn’s disease (42%) and ulcerative colitis (40%) and uncommon (10%) in 79 patients with newly diagnosed coeliac disease. In Crohn’s disease, concurrent Fructose malabsorption and lactose malabsorption was most common (29%), and the association of fructose malabsorption with ERBHAL seen overall (62%) was not observed (36%, P < 0.0001).

Conclusions  Carbohydrate malabsorption and ERBHAL are normal physiological phenomena. The abnormal patterns observed in Crohn’s disease may have pathogenic importance.
Risks and benefits of splenectomy versus no splenectomy for hereditary spherocytosis – a personal view
Authors: Schilling Robert F.
Published: April 2009

Splenectomy is indicated in hereditary spherocytosis to relieve symptoms due to anaemia or splenomegaly, reverse growth failure or skeletal changes due to over‐robust erythropoiesis, and prevent recurrent gallstones. A life‐long risk of bacterial infection has been recognised for many years as a concomitant cost of splenectomy. The scope of this risk has expanded to include a number of organisms beyond the triad of pneumococcus, meningococcus, and haemophilus influenzae. Recently, it has been demonstrated that splenectomy also confers a significant risk of delayed adverse vascular events in patients with hereditary spherocytosis, just as it does in patients undergoing splenectomy for other indications. Further, these same studies demonstrated a benefit of avoiding splenectomy: hereditary spherocytosis patients with a spleen have significantly fewer adverse vascular events than unaffected family members, probably because of the protective effect of chronic, mild anaemia. Accordingly, this review marshals the evidence favouring a conservative approach to splenectomy in spherocytosis.
Immunomodulation of innate immune responses by vasoactive intestinal peptide (VIP): its therapeutic potential in inflammatory disease
Authors: Smalley S G R , Barrow P A , Foster N
Published: April 2009

Since the late 1970s a number of laboratories have studied the role of vasoactive intestinal peptide (VIP) in inflammation and immunity. These studies have highlighted the dramatic effect of VIP on immune cell activation and function, and studies using animal models of disease have indicated that VIP has significant therapeutic and prophylactic potential. This review will focus on the effects of VIP on innate immune cell function and discuss the therapeutic potential for VIP in inflammatory diseases of humans.
Histamine H1-receptor antagonists with immunomodulating activities: potential use for modulating T helper type 1 (Th1)/Th2 cytokine imbalance and inflammatory responses in allergic diseases
Authors: Okamoto T , Iwata S , Ohnuma K , Dang N H , Morimoto C
Published: April 2009

Being a first-line treatment for hypersensitivity allergic disease, histamine H1-receptor antagonists possess anti-inflammatory activity in addition to being H1-receptor antagonists. While it is not purely a histamine-related condition, hypersensitivity allergic disease is associated with an increase in the number of T helper type 2 (Th2) cells and Th2 cytokines, and a decrease in the number of Th1 cells and Th1 cytokines. Suppression of Th2-type cytokine production in addition to H1-receptor blockade may therefore represent a successful therapeutic strategy for the treatment of hypersensitivity allergic diseases. H1-receptor antagonists have been reported to modulate immune cascade at various points by acting on T cell-related inflammatory molecules, including adhesion molecules, chemokines and inflammatory cytokines. These effects of H1-receptor antagonists may be optimized for the treatment of allergic diseases. Besides their ability to regulate inflammatory molecules, some H1-receptor antagonists have been reported to down-regulate Th2 cytokine production. In particular, it has been shown that several H1-receptor antagonists specifically inhibit the production of Th2, but not Th1, cytokines. Accumulating evidence indicates a crucial role for Th1/Th2 cytokine imbalance on the development of allergic diseases. Accordingly, the use of H1-receptor antagonist with Th2 cytokine inhibitory activity to modulate Th1/Th2 cytokine imbalance might be a favourable strategy for the treatment of hypersensitivity allergic diseases. Furthermore, the identification of H1-receptor antagonists which possess immunoregulatory activities in addition to their anti-histamine activity will provide an important insight into the development of novel immunoregulatory drugs.
Enhanced complement activation is part of the unfavourable cardiovascular risk profile in South Asians
Authors: Siezenga M A , Chandie Shaw P K , Van Der Geest R N , Mollnes T E , Daha M R , Rabelink T J , Berger S P
Published: April 2009

South Asian immigrants in western societies exhibit a high burden of diabetes and subsequent vascular complications. Diabetic vascular complications are associated with vascular inflammation. We hypothesize that enhanced complement activation is involved. Therefore, levels of complement C3 and SC5b-9 – the soluble end product of complement activation – in a group of 200 South Asians were compared with an age- and sex-matched control group of native Caucasians. In addition, the association between complement levels and albuminuria, an indicator of renal damage and a cardiovascular risk marker, was assessed in the diabetic South Asian group. Compared with native Caucasians, South Asians had significantly higher levels of both serum C3 and plasma SC5b-9, even when only non-diabetic South Asians were considered. Diabetic South Asians had significantly higher C3 levels compared with non-diabetic South Asians. In diabetic South Asians, higher levels of SC5b-9 were associated with an increased prevalence of albuminuria (odds ratio 5·4, 95% confidence interval 1·8–15·8). These results suggest that enhanced complement activation is part of the unfavourable cardiovascular risk profile in South Asians.
Direct evidence for roles of phosphorylated regulatory light chain of myosin II in furrow ingression during cytokinesis in HeLa cells
Authors: Asano Satoshi , Hamao Kozue , Hosoya Hiroshi
Published: April 2009

Phosphorylation of myosin II is thought to play an important role in cytokinesis. Although it is well known that phosphorylated regulatory light chain of myosin II (P‐MRLC) localizes along the contractile ring, it is not clear how P‐MRLC controls myosin II and F‐actin in furrow ingression during cytokinesis. To elucidate roles of P‐MRLC in furrow ingression, HeLa cells transfected with EGFP‐tagged wild‐type or each MRLC mutant were observed using a live‐imaging microscope. Time‐lapse observation revealed that a delay of furrow ingression was observed in the nonphosphorylatable form of MRLC (AA‐MRLC)‐expressing cell but not in the wild‐type or phospho‐mimic MRLC‐expressing cell. Among each form of MRLC‐expressing cell, the total amount of P‐MRLC including phospho‐mimic MRLCs was smallest in the cell expressing AA‐MRLC. However, the amount of F‐actin and myosin II at the contractile ring in the AA‐MRLC‐expressing cell was the same as that in the normal cell. Interestingly, delay of furrow ingression by a Rho‐kinase inhibitor, Y27632, was rescued by phospho‐mimic MRLCs. These results suggest that the P‐MRLC is essential for the progress of furrow ingression but not the retainment of F‐actin and myosin II in the contractile ring of dividing HeLa cells.
Schizosaccharomyces pombe cell division cycle under limited glucose requires Ssp1 kinase, the putative CaMKK, and Sds23, a PP2A‐related phosphatase inhibitor
Authors: Hanyu Yuichiro , Imai Kumiko K. , Kawasaki Yosuke , Nakamura Takahiro , Nakaseko Yukinobu , Nagao Koji , Kokubu Aya , Ebe Masahiro , Fujisawa Asuka , Hayashi Takeshi , Obuse Chikashi , Yanagida Mitsuhiro
Published: April 2009

Calcium/calmodulin‐dependent protein kinase (CaMK) is required for diverse cellular functions, and similar kinases exist in fungi. Although mammalian CaMK kinase (CaMKK) activates CaMK and also evolutionarily‐conserved AMP‐activated protein kinase (AMPK), CaMKK is yet to be established in yeast. We here report that the fission yeast Schizosaccharomyces pombe Ssp1 kinase, which controls G2/M transition and response to stress, is the putative CaMKK. Ssp1 has a CaM binding domain (CBD) and associates with 14‐3‐3 proteins as mammalian CaMKK does. Temperature‐sensitive ssp1 mutants isolated are defective in the tolerance to limited glucose, and this tolerance requires the conserved stretch present between the kinase domain and CBD. Sds23, multi‐copy suppressor for mutants defective in type 1 phosphatase and APC/cyclosome, also suppresses the ssp1 phenotype, and is required for the tolerance to limited glucose. We demonstrate that Sds23 binds to type 2A protein phosphatases (PP2A) and PP2A‐related phosphatase Ppe1, and that Sds23 inhibits Ppe1 phosphatase activity. Ssp1 and Ppe1 thus seem to antagonize in utilizing limited glucose. We also show that Ppk9 and Ssp2 are the catalytic subunits of AMPK and AMPK‐related kinases, respectively, which bind to common β‐(Amk2) and γ‐(Cbs2) subunits.
Growth rate dependent numbers of SeqA structures organize the multiple replication forks in rapidly growing Escherichia coli
Authors: Morigen  , Odsbu Ingvild , Skarstad Kirsten
Published: April 2009

When the bacterium Escherichia coli is grown in rich medium, the replication and segregation periods may span two, three or four generations and cells may contain up to 24 replication forks. The newly synthesized, hemimethylated DNA at each fork is bound by SeqA protein. The SeqA–DNA structures form distinct foci that can be observed by immunofluorescence microscopy. The numbers of foci were lower than the numbers of replication forks indicating fork co‐localization. The extent of co‐localization correlated with the extent of replication cycle overlap in wild‐type cells. No abrupt increase in the numbers of foci occurred at the time of initiation of replication, suggesting that new replication forks bind to existing SeqA structures. Manipulations with replication control mechanisms that led to extension or reduction of the replication period and number of forks, did not lead to changes in the numbers of SeqA foci per cell. The results indicate that the number of SeqA foci is not directly governed by the number of replication forks, and supports the idea that new DNA may be ‘captured’ by existing SeqA structures.
Ecological specialization of the aphid Aphis gossypii Glover on cultivated host plants
Authors: CARLETTO J. , LOMBAERT E. , CHAVIGNY P. , BRÉVAULT T. , LAPCHIN L. , VANLERBERGHE‐MASUTTI F.
Published: April 2009

Many plant‐feeding insect species considered to be polyphagous are in fact composed of genetically differentiated sympatric populations that use different hosts and between which gene flow still exists. We studied the population genetic structure of the cotton‐melon aphid Aphis gossypii that is considered as one of the most polyphagous aphid species. We used eight microsatellites to analyse the genetic diversity of numerous samples of A. gossypii collected over several years at a large geographical scale on annual crops from different plant families. The number of multilocus genotypes detected was extremely low and the genotypes were found to be associated with host plants. Five host races were unambiguously identified (Cucurbitaceae, cotton, eggplant, potato and chili‐ or sweet pepper). These host races were dominated by asexual clones. Plant transfer experiments using several specialized clones further confirmed the existence of host‐associated trade‐offs. Finally, both genetic and experimental data suggested that plants of the genus Hibiscus may be used as refuge for the specialized clones. Resource abundance is discussed as a key factor involved in the process of ecological specialization in A. gossypii.
The PFKFB3 splice variant UBI2K4 is downregulated in high‐grade astrocytomas and impedes the growth of U87 glioblastoma cells
Authors: Zscharnack K. , Kessler R. , Bleichert F. , Warnke J. P. , Eschrich K.
Published: April 2009

Aims: Fructose‐2,6‐bisphosphate, a key regulator of glycolysis, is synthesized and degraded by four different isozymes of 6‐phosphofructo‐2‐kinase/fructose‐2,6‐bisphosphatase (PFKFB1–4). The PFKFB3 isozyme is upregulated in several human tumours. Six alternatively spliced variants of PFKFB3 mRNA are known in humans (UBI2K1–6). Here, we studied the role of the PFKFB3 splice variants in human astrocytic gliomas. Methods: We analysed the PFKFB3 splice variants in 48 astrocytic gliomas by RT‐PCR and real‐time PCR. The effect of transient and stable overexpression of the PFKFB3 isoforms was studied in U87 glioblastoma cells by MTT, cell counting, clone formation assay and metabolic measurements. Results: UBI2K5 and UBI2K6 are the predominant splice variants in rapidly proliferating high‐grade astrocytomas while the expression of UBI2K3 and UBI2K4 is mainly restricted to low‐grade astrocytomas and nonneoplastic brain tissue. Overexpression of UBI2K5 or UBI2K6 in the U87 glioblastoma cell line enhances the glycolytic flux but does not affect cell growth. In contrast, overexpression of UBI2K4 reduces cell viability and anchorage‐independent growth of U87 cells. The UBI2K4 mRNA level is downregulated in astrocytic gliomas with increasing malignancy grade. Moreover, the UBI2K4 mRNA level correlates with growth rate of several human cancer cell lines derived from different tissue types. Conclusions: Our results demonstrate that the splice variant UBI2K4 impedes the tumour cell growth and might serve as a tumour suppressor in astrocytic tumours.
Factors that affect hearing level in individuals with the mitochondrial 1555A>G mutation
Authors: Lu SY , Nishio S , Tsukada K , Oguchi T , Kobayashi K , Abe S , Usami S
Published: April 2009

The mitochondrial 1555A>G mutation is one of the most common mutations responsible for hearing loss in Asians. Although the association with aminoglycoside exposure is well known, there is great variation in the severity of hearing loss. We analyzed hearing levels in 221 Japanese individuals with this mutation and attempted to identify relevant covariants including (i) age, (ii) aminoglycoside exposure, (iii) heteroplasmy ratio, and (iv) other gene mutations. At every age, average hearing levels were worse than those in normal subjects, suggesting that mitochondrial function itself may affect the severity of hearing loss. Although the hearing loss in individuals with the 1555A>G mutation progressed with age, the rate did not differ from that of the normal subjects. Those who had reported aminoglycoside exposure had moderate‐to‐severe hearing impairment regardless of age, confirming that such exposure is the most important environmental variable. We also confirmed the presence of heteroplasmy, which is known to modify the expression of other mitochondrial diseases, but found no evidence for a significant correlation with hearing impairment. A high prevalence of GJB2 heterozygous mutations was noted, indicating that these mutations may exhibit epistatic interaction with the 1555A>G mutation.
CASK: a scaffold protein involved in X‐linked brain malformation phenotypes
Authors: Sanders SS
Published: April 2009

Mutations of CASK cause an X‐linked brain malformation phenotype with microcephaly and hypoplasia of the brainstem and cerebellum Najm et al. (2008) Nature Genetics 40: 1065–1067
Recurrence risk due to germ line mosaicism: Duchenne and Becker muscular dystrophy
Authors: Helderman‐van den Enden ATJM , De Jong R , Den Dunnen JT , Houwing‐Duistermaat JJ , Kneppers ALJ , Ginjaar HB , Breuning MH , Bakker E
Published: April 2009

The presence of multiple affected offspring from apparently non‐carrier parents is caused by germ line mosaicism. Although germ line mosaicism has been reported for many diseases, figures for recurrence risks are known for only a few of them. In X‐linked Duchenne and Becker muscular dystrophies (DMD/BMD), the recurrence risk for non‐carrier females due to germ line mosaicism has been estimated to be between 14% and 20% (95% confidence interval 3–30) if the risk haplotype is transmitted. In this study, we have analyzed 318 DMD/BMD cases in which the detected mutation was de novo with the aim of obtaining a better estimate of the ‘true’ number of germ line mosaics and a more precise recurrence risk. This knowledge is essential for genetic counseling. Our data indicate a recurrence risk of 8.6% (4.8–12.2) if the risk haplotype is transmitted, but there is a remarkable difference between proximal (15.6%) (4.1–27.0) and distal (6.4%) (2.1–10.6) deletions. Overall, most mutations originated in the female. Deletions occur more often on the X chromosome of the maternal grandmother, whereas point mutations occur on the X chromosome of the maternal grandfather. In unhaplotyped de novo DMD/BMD families, the risk of recurrence of the mutation is 4.3%.
A prospective study examining the effects of gender and sexual/physical abuse on mood outcomes in patients with co‐occurring bipolar I and substance use disorders
Authors: Meade Christina S , McDonald Leah J , Graff Fiona S , Fitzmaurice Garrett M , Griffin Margaret L , Weiss Roger D
Published: April 2009

Objectives:  Prior research suggests possible gender differences in the longitudinal course of bipolar disorder. This study prospectively examined gender differences in mood outcomes and tested the effects of sexual/physical abuse and posttraumatic stress disorder (PTSD).

Methods:  Participants (49 men, 41 women) with co‐occurring bipolar I and substance use disorders (92% alcohol, 42% drug) were enrolled in a group treatment trial. They were followed for eight months, with monthly assessments, yielding 32 weeks of data. Primary outcome measures were number of weeks in each mood state, recurrences of depression or mania, and polarity shifts from depression to mania or vice versa. Negative binomial regression was used to examine the effects of gender, lifetime abuse, and PTSD on these outcomes.

Results:  Participants met syndromal criteria for a mood episode on a mean of 27% of 32 weeks, with depression occurring most frequently. Compared to men, women reported significantly more weeks of mixed mania [relative rate (RR) = 8.53], fewer weeks of euthymia (RR = 0.58), more recurrences of mania (RR = 1.96), and more direct polarity shifts (RR = 1.49) (all p < 0.05). Women also reported significantly higher rates of lifetime sexual or physical abuse (68% versus 33%), which partially explained the relationships between gender and mixed mania and direct polarity shifts.

Conclusions:  Participants experienced persistent mood symptoms over time. Women consistently reported poorer mood outcomes, and lifetime abuse may help explain observed gender differences in mood outcomes. Further research is necessary to better understand the treatment implications of these findings.
Long‐term physical outcome in patients with septic shock
Authors: POULSEN J. B. , MØLLER K. , KEHLET H. , PERNER A.
Published: April 2009

Background: Limited information is available on physical function after septic shock. The aim of the present study was to assess the physical outcome in survivors 1 year after septic shock.

Methods: The outcome status of all 174 adult patients admitted to a mixed ICU with the diagnosis septic shock in a 1‐year period was registered. Survivors were interviewed about physical function and socioeconomic status using a questionnaire including the Short Form‐36 survey. The pre‐ICU‐admission Functional Comorbidity Index (FCI) was also registered.

Results: Of the 80 survivors, two were still hospitalised; thus, 78 were invited to participate and 70 replied (inclusion‐rate 88%). Patients were followed up at median 351 days after hospital discharge. At follow‐up the patients had a markedly reduced physical component summary score (PCS) compared with age‐ and sex‐adjusted general population controls (36 vs. 47, P<0.0001). This was also observed in patients with no comorbidity before ICU admission (34 vs. 47, P<0.001). There was a negative correlation (r=−0.27, P=0.03) between pre‐ICU‐admission FCI values and the PCS at follow‐up. According to 81% of the patients, loss of muscle mass was the main cause of decreased physical function. Only 43% (10 vs. 23, P=0.01) of the previously employed had returned to work, and the number of patients in need of home‐based personal assistance had doubled (10/20, P=0.04).

Conclusion: Physical function is substantially reduced in survivors of septic shock 1 year after discharge.
Continuous femoral nerve block after total knee arthroplasty?
Authors: KADIC L. , BOONSTRA M. C. , DE WAAL MALEFIJT M. C. , LAKO S. J. , VAN EGMOND J. , DRIESSEN J. J.
Published: April 2009

Background: A continuous femoral nerve block is frequently used as an adjunct therapy after total knee arthroplasty (TKA). However, there is still debate on its benefits.

Methods: In this prospective, randomized study, patients received a basic analgesic regimen of paracetamol and dicloflenac for the first 48 h postoperatively. In addition, the study group received a continuous femoral nerve block. A morphine patient‐controlled analgesia pump was also available as a rescue analgesic to all the patients. Patients' numeric rating scores for pain, the amount of morphine consumed and its side effects during the first 48 h were recorded. Knee flexion angles achieved during the first week were registered. Three months postoperatively, patients completed Western Ontario and McMaster Universities Osteoarthritis Index and Knee Society Score.

Results: The study group (n=27) had less pain (P=0.0016) during the first 48 h, was more satisfied with the analgesia (P<0.001) and used less morphine (P=0.007) compared with the control group (n=26). Fewer patients were nauseated, vomited or were drowsy in the study group (P=0.001). Also, the study group achieved better knee flexion in the first 6 days after surgery (P=0.001), with more patients reaching 90° flexion than the control group. However, after 3 months, there were no significant functional differences between the groups.

Conclusion: A continuous femoral nerve block leads to better analgesia, less morphine consumption and less morphine‐related side effects after TKA. Early functional recovery is improved, resulting in more patients reaching 90° knee flexion after 6 days. However, after 3 months, no significant functional benefits were found.