bluuebunny/crossref_metadata_2025 huggingface dataset:
Genes associated with the formation of germ cells from embryonic stem cells in cultures containing different glucose concentrations
Authors: Mizuno Kentaro , Tokumasu Ako , Nakamura Aya , Hayashi Yutaro , Kojima Yoshiyuki , Kohri Kenjiro , Noce Toshiaki
Published: January 2006

In a previous study, we established a system for visualizing the development of germ cells from mouse embryonic stem (ES) cells in culture using knock‐in ES clones in which visual reporter genes were expressed from the mouse vasa homolog, Mvh. While assessing various culture conditions, we found that germ‐cell formation was markedly depressed in low glucose medium. Using a repeated polymerase chain reaction (PCR) subtraction method, we identified genes that were differentially expressed in low versus high glucose media. Three genes that were predominantly expressed in high glucose medium, thioredoxin‐interacting protein (Txnip), pituitary tumor‐transforming gene 1 (Pttg), and RuvB‐like protein 2 (RuvBl2), were further investigated. These genes were also found to be highly expressed in adult and embryonic gonads, and RuvBl2 in particular, which encodes an ATP‐dependent DNA helicase, was specifically detected in the spermatocytes and spermatids of the adult testis as well as in primordial germ cells. Furthermore, using a green fluorescent protein (GFP) fusion construct, we found that RuvBl2 was expressed in both the nucleus and cytoplasm of testicular germ cells. These findings suggest a possible relationship between glucose metabolism and germ‐cell development. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Characterization of vitellogenin in the shrimp Metapenaeus ensis: Expression studies and hormonal regulation of MeVg1 transcription in vitro
Authors: Tiu Shirley H.K. , Hui Jerome H.L. , HE Jian‐Guo , Tobe Stephen S. , Chan Siu‐Ming
Published: January 2006

During gonad maturation, female shrimp accumulate the major egg yolk protein vitellin (Vn) in premolt stage, and the process of molting and reproduction is synchronized. Using a polyclonal anti‐Vn antibody, immunopositive signals could be detected in the ovary and among the proteins secreted by the hepatopancreas by Western blot. In the ovary, Vn immunoreactivity was located in the posterior lobe. Hepatopancreas proteins with sizes identical to ovary vitellogenin (Vg) subunits (i.e., 78 and 157 kDa) were immunoreactive to the Vn antibody and these proteins included amino acid sequences identical to parts of the MeVg1 precursor. A major 7.8 kb MeVg1 transcript, was detected in the ovary. In the hepatopancreas, the transcripts were primarily small (<2.3 kb) and while the 7.8 kb transcript which constitutes <50% of the total Vg mRNA. MeVg1 transcript could be detected in the hepatopancreas of juvenile females with a maximum level during late intermolt and early premolt. To study the effect of different hormones on expression of MeVg1, explant cultures of hepatopancreas and ovary were developed. Although several hormones (20‐hydroxyecdysone, estradiol (ES), farnesoic acid (FA), juvenile hormone (JH) III, methyl farnesoate, and progesterone (PG)) apparently stimulated MeVg1 gene expression, only FA consistently stimulated MeVg1 expression by the hepatopancreas explants, while both FA and 20‐hydroxyecdysone were stimulated ovarian explants. In summary, (i) Vg transcripts can be detected in both reproductive and nonreproductive females; (ii) the presence of large quantities of smaller Vg transcripts and the absence of a large Vg precursor from the hepatopancreas suggests that smaller MeVg1 transcripts provide an important contribution to Vg synthesis in shrimp. Our results suggest that there is differential processing of the MeVg1 precursor in the ovary and hepatopancreas of shrimp. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Chimeric honeybees (Apis mellifera) produced by transplantation of embryonic cells into pre‐gastrula stage embryos and detection of chimerism by use of microsatellite markers
Authors: Bergem M. , Norberg K. , Roseth A. , Meuwissen T. , Lien S. , Aamodt R. H.
Published: January 2006

The production of chimeras, by use of cell transplantation, has proved to be highly valuable in studies of development by providing insights into cell fate, differentiation, and developmental potential. So far, chimeric honeybees have been created by nuclear transfer technologies. We have developed protocols to produce chimeric honeybees by use of cell transplantation. Embryonic cells were transplanted between pre‐gastrula stage embryos (32–34 hr after oviposition) and hatched larvae were reared in vitro for 4 days. Chimeric individuals were detected by use of microsatellite analysis and a conservative estimation approach. 4.8% of embryos, posteriorly injected with embryonic cells, developed into chimeric honeybee larvae. By injection of cells pre‐stained with fluorescent cell tracer dye, we studied the integration of transplanted cells in the developing embryos. Number of injected cells varied from 0 to 50 and cells remained and multiplied mainly in the area of injection. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Denuding and centrifugation of maturing bovine oocytes alters oocyte spindle integrity and the ability of cytoplasm to support parthenogenetic and nuclear transfer embryo development
Authors: Li Guang‐Peng , Bunch Thomas D. , White Kenneth L. , Rickords Lee , Liu Ying , Sessions Benjamin R.
Published: January 2006

The effects of cumulus cell removal and centrifugation of maturing bovine oocytes on nuclear maturation and subsequent embryo development after parthenogenetic activation and nuclear transfer were examined. Removal of cumulus cells at 4, 8, and 15 hr after in vitro maturation (IVM) or the centrifugation of denuded oocytes had no effect on maturation rates. Oocytes treated at 0 hr of IVM had a lower expulsion rate (50%) of the first polar body (PB1). The removal of cumulus cells and centrifugation affected the pattern of spindle microtubule distribution and division of chromosomes. There were almost no spindle microtubules allocated to PB1 and the spindles were swollen in anaphase I and telophase I oocytes. Approximately 20% of PB1 oocytes contained tripolar or multipolar spindles. After activation, oocytes denuded with or without centrifugation at 8 hr of IVM resulted in the lowest rate of development (3.0%). Denuded oocytes at 4, 15, and 24 hr of IVM with centrifugation or not resulted in similar blastocyst development rates (9.6%–13.2%). However, centrifugation of oocytes denuded at the beginning of IVM resulted in lower blastocyst development rate (8.1%, P < 0.05) than the noncentrifuged oocytes (17.3%). After nuclear transfer, the blastocyst development rates of oocytes denuded and centrifuged at 0, 4, and 8 hr of IVM were not different when compared to the same patch of noncentrifuged oocytes. However, oocytes denuded and centrifuged at 15 hr of IVM resulted in lower (P < 0.05) blastocyst development rates than the noncentrifuged oocytes. The results of this study suggest that removal of cumulus cells and centrifugation of denuded oocytes affect the spindle pattern. Embryo development of denuded and centrifuged oocytes may differ depending on the time of removal of cumulus cells. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Expression of p27BBP/eIF6 is highly modulated during Xenopus laevis embryogenesis
Authors: Vaccaro Maria Carmela , Cuccaro Marta , Marco Nadia De , Campanella Chiara
Published: January 2006

Protein p27BBP/eIF6 is necessary for ribosomal function of all cells. Previous data showed that from mammals to yeast p27BBP/eIF6 is involved in the biogenesis of ribosomal subunit 60S and its association with the 60S prevents premature 80S formation regulated by PKC signaling, indicating that phosphorylation of p27BBP/eIF6 is needed for translation to occur. While in vitro p27BBP/eIF6 is constitutively expressed, and it has a high level of expression in cycling cells, in vivo its expression varies according to tissues and appears regulated by factors up to now unknown. p27BBP/eIF6 has never been investigated in developing organisms where its upregulation can be correlated with tissue growth and differentiation. In this study we have sequenced p27BBP/eIF6 cDNA and studied its expression during development of Xenopus laevis, as the first step for studying its regulation. The amino acid sequence is highly conserved with two putative PKC phosphorylation sites in serine, one site being typical of Xenopus. At the end of gastrulation, the p27BBP/eIF6 riboprobe localizes in the neural plate and in the paraxial mesoderm. In particular, from stage 24, a clear‐cut localization occurs in the perspective head. In embryos exposed to teratogens, the localization of p27BBP/eIF6 riboprobe varies according to the change of head size caused by the treatment. p27BBP/eIF6 expression is particularly evident in differentiating olfactory pits, the lens, otic vesicles, and in branchial arches. Features of particular interest are p27BBP/eIF6 high level of expression in the eye field, and in the mid‐hindbrain‐boundary, two regions with high proliferative activity. Altogether, data indicate that a modulated expression of p27BBP/eIF6 occurs in developing anlagens in addition to a basal level of expression, and may suggest a correlation between p27BBP/eIF6 and proliferative activity. Moreover, the X. laevis cDNA isolation and characterization offer new hints for further studies in relation to potential p27BBP/eIF6 phosphorylation. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Effects of imatinib mesylate (Glivec®) as a c‐kit tyrosine kinase inhibitor in the guinea‐pig urinary bladder
Authors: Kubota Yasue , Biers Suzanne M. , Kohri Kenjiro , Brading Alison F.
Published: January 2006

Aims

In the gastrointestinal tract, slow wave activity in smooth muscle is generated by the interstitial cells of Cajal (ICC). Detrusor smooth muscle strips of most species show spontaneous contractions which are triggered by action potential bursts, however, the pacemaker mechanisms for the detrusor are still unknown. Recently, ICC‐like cells have been found in guinea‐pig bladder, using antibodies to the c‐kit receptor. We have investigated the effects of Glivec, a c‐kit tyrosine kinase inhibitor, on spontaneous action potentials in guinea‐pig detrusor and intravesical pressure of isolated guinea‐pig bladders.

Methods

Changes in the membrane potential were measured in guinea‐pig detrusor smooth muscle using conventional microelectrode techniques. Pressure changes in the bladder were recorded using whole organ bath techniques.

Results

Smooth muscle cells in detrusor muscle bundles exhibited spontaneous action potentials, and spontaneous pressure rises occurred in isolated bladders. Glivec (10 μM) converted action potential bursts into continuous firing with no effects on the shape of individual action potentials. Glivec (>50 μM) reduced the amplitude of spontaneous pressure rises in the whole bladder in a dose dependent manner and abolished spontaneous action potentials in detrusor smooth muscle cells.

Conclusions

The results suggest that ICC‐like cells may be responsible for generating bursts of action potentials and contractions in detrusor smooth muscle. Drugs inhibiting the c‐kit receptor may prove useful for treating the overactive bladder. Neurourol. Urodynam. 25:205–210, 2006. © 2006 Wiley‐Liss, Inc.
Lipid signaling changes in smooth muscle remodeling associated with partial urinary bladder outlet obstruction
Authors: LaBelle Edward , Zderic Stephen , Delaney Daniel , Hypolite Joseph , Wein Alan , Chacko Samuel
Published: January 2006

Aims

Hypertrophy of the urinary bladder smooth muscle (detrusor) is associated with partial bladder outlet obstruction (PBOO). Hypertrophied detrusor smooth muscle (DSM) reveals altered contractile characteristics. In this study, we analyzed the lipid‐dependent signaling system that includes phospholipase A2in PBOO‐induced DSM remodeling and hypertrophy to determine whether the release of arachidonic acid (AA) from phospholipid is altered in the detrusor.

Methods

Partial bladder outlet obstruction (PBOO) was produced by partial ligation of the urethra in New Zealand white rabbits. Two weeks after the surgery, the bladder function was studied by keeping the rabbits in metabolic cages for 24 hr. Bladders were removed from rabbits that had bladder dysfunction (increased urinary frequency and decreased void volume) and the DSM separated from mucosa and serosa. The isolated smooth muscle was incubated with [3H] AA to equilibrate the cytoplasmic AA. The level of AA release was compared with the level obtained with 2‐week sham‐operated rabbits.

Results

The rate of AA release was high in DSM from bladders with PBOO‐induced hypertrophy. Carbachol stimulated AA release in control DSM but DSM from obstructed rabbits revealed no further increase from the elevated basal AA release. The half‐maximal concentration of carbachol that was required to stimulate AA release from control samples of detrusor was 35 µM.

Conclusions

The increased levels of AA release that are observed in this tissue after PBOO indicate the activation of phospholipase A2. The finding that carbachol could induce contraction, but not an increase in AA, indicates that the carbachol‐induced contraction in the obstructed bladders is independent of lipid signaling pathways that involve AA. It is possible that the increased rate of arachidonic acid release from obstructed bladders correlates with the enhanced rates of prostaglandin production reported by other investigators from the same tissue. Neurourol. Urodynam. © 2006 Wiley‐Liss, Inc.
Analysis of biologic surrogate markers from a Children's Oncology Group Phase I trial of gefitinib in pediatric patients with solid tumors
Authors: Jimeno Antonio , Daw Najat C. , Amador Maria L. , Cusatis George , Kulesza Peter , Krailo Mark , Ingle Ashish M. , Blaney Susan M. , Adamson Peter , Hidalgo Manuel
Published: January 2006

This trial evaluated the effect of gefitinib on the plasma circulating levels of epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)‐2 and ‐9 of patients treated on a pediatric Phase I trial. Complete plasma correlative studies were obtained from 16 of the 25 enrolled patients. There was a trend for lower MMP‐2 baseline levels in patients with partial response or stable disease. The Ewing sarcoma from the only patient with partial response lacked egfr mutations. Gefitinib did not induce any significant variation in the levels of the assessed parameters, and none of these determinations showed significant predictive or prognostic value. Pediatr Blood Cancer 2007;49:352–357. © 2006 Wiley‐Liss, Inc.
The outcome of children with acute myeloid leukemia (AML) post‐allogeneic stem cell transplantation (SCT) is not improved by the addition of etoposide to the conditioning regimen
Authors: Ayas Mouhab , Al‐Seraihi Amal , Al‐Mahr Mohamad , Al‐Jefri Abdullah , Belgaumi Assim , Elhassan Ibrahim , El‐Solh Hassan
Published: January 2006

Background

Relapse remains a concern for children with AML undergoing allogeneic SCT, so in an effort to reduce the risk of relapse in these patients, we intensified our pre‐SCT preparation by adding etoposide to the standard busulfan and cyclophosphamide regimen.

Procedure

We retrospectively analyzed the collected data and compared the two groups; Group A (n = 18) included patients who received busulfan 16 mg/kg plus cyclophosphamide 200 mg/kg (Bu/Cy), and Group B (n = 48) included patients who received busulfan 12 mg/kg, cyclophosphamide 90 mg/kg in addition to etoposide 60 mg/kg (Bu/Cy/VP). The patients' characteristics were similar in the two groups.

Results

No significant difference in the overall outcome was noted; the 5‐year overall survival was 50% and 53.3% for Groups A and B, respectively (P = 0.9). Similarly, the 5‐year probability of relapse was 64.1% and 46.1% for Groups A and B, respectively (P = 0.38). The use of etoposide was not associated with increased toxicity.

Conclusion

The addition of etoposide to the Bu/Cy regimen was well tolerated, but did not appear to improve the outcome. Pediatr Blood Cancer © 2006 Wiley‐Liss, Inc.
The proteomic analysis of an adipocyte differentiated from human mesenchymal stem cells using two‐dimensional gel electrophoresis
Authors: Lee Hyun‐Kyung , Lee Byung‐Hyo , Park Seung‐Ah , Kim Chan‐Wha
Published: January 2006

Adipose tissues play a crucial endocrine role in the control of whole body glucose homeostasis and insulin sensitivity. Considering the current substantial rise in obesity and obesity‐related diseases, including diabetes, it is important to understand the molecular basis of adipocyte differentiation and its control. In this study, we have analyzed the protein expression inherent to adipogenic differentiation, by 2‐DE, MALDI‐TOF, and RT‐PCR. This study focused on proteins that were differentially expressed by the differentiation of human mesenchymal stem cells (hMSCs) to adipocytes. We conducted 2‐DE for each set of proteins in the cytosol of adipocytes that had differentiated from hMSC, in a pH range from 3–10. Thirty‐two protein spots were shown to have different expression levels. Among these, eight up‐regulated proteins were identified by MALDI‐TOF/MS, as the following: syntaxin binding protein 3, OSBP‐related protein 3, phosphodiesterase, glycophorin, immunoglobulin kappa chain variable region, peroxisome proliferative activated receptor gamma (PPAR‐γ), bA528A10.3.1 (novel protein similar to KIAA01616, isoform 1), and T cell receptor V‐β 4. Four proteins: syntaxin‐3, OSBP‐related protein 3, PPAR‐γ and glycophorin were associated with adipogenesis.
Proteomic analysis of heterotrophy in Synechocystis sp. PCC 6803
Authors: Kurian Dominic , Jansèn Tove , Mäenpää Pirkko
Published: January 2006

To provide an insight into the heterotrophic metabolism of cyanobacteria, a proteomic approach has been employed with the model organism Synechocystis sp. PCC 6803. The soluble proteins from Synechocystis grown under photoautotrophic and light‐activated heterotrophic conditions were separated by 2‐DE and identified by MALDI‐MS or LC‐MS/MS analysis. 2‐DE gels made using narrow‐ and micro‐range IPG strips allowed quantitative comparison of more than 900 spots. Out of 67 abundant protein spots identified, 13 spots were increased and 9 decreased under heterotrophy, representing all the major fold changes. Proteomic alterations and activity levels of selected enzymes indicate a shift in the central carbon metabolism in response to trophic change. The significant reduction in light‐saturated rate of photosynthesis as well as in the expression levels of rubisco and CO2‐concentrating mechanism proteins under heterotrophy indicates the down‐regulation of the photosynthetic machinery. Alterations in the expression level of proteins involved in carbon utilization pathways refer to enhanced glycolysis, oxidative pentose phosphate pathway as well as tricarboxylic acid cycle under heterotrophy. Proteomic evidences also suggest an enhanced biosynthesis of amino acids such as histidine and serine during heterotrophic growth.
Tumor‐related immunity in prostate cancer patients treated with human recombinant granulocyte monocyte‐colony stimulating factor (GM‐CSF)
Authors: Schwaab Thomas , Tretter Christopher P.G. , Gibson Jennifer J. , Cole Bernard F. , Schned Alan R. , Harris Robert , Fisher Jan L. , Crosby Nancy , Stempkowski Laura M. , Heaney John A. , Ernstoff Marc S.
Published: January 2006

BACKGROUND

Granulocyte monocyte‐colony stimulating factor (GM‐CSF) supports the survival, expansion, and differentiation of lymphoid and myeloid derived dendritic cells (DCs). We hypothesized that systemic therapy with GM‐CSF in prostate cancer patients could augment prostate cancer‐related immunity and induce clinical response.

METHODS

Eligible patients were randomly assigned to receive either 125 or 250 µg/m2 GM‐CSF subcutaneously three times a week until clinical progression. Prostate‐specific antigen (PSA) T cell precursor frequencies were determined by a flow cytometric method.

RESULTS

We were able to show, for the first time, a statistically significant correlation between pre‐treatment PSA level and PSA‐specific CD4+ T cell precursors and a trend between pre‐treatment PSA level and PSA‐specific CD8+ T cell precursors (P <0.0001 and P = 0.059, respectively).

CONCLUSIONS

These results suggest that existent immunity to PSA in prostate cancer patients may be a promising target for future immunotherapeutic approaches to prostate cancer. Prostate 66:667–674, 2006. © 2006 Wiley‐Liss, Inc.
Differentiation‐dependent expression of hypothetical proteins in the neuroblastoma cell line N1E‐115
Authors: Oh Ji‐eun , Karlmark Karlin Raja , Shin Jooho , Hengstschläger Markus , Lubec Gert
Published: January 2006

Several protein cascades, including signaling, cytoskeletal, chaperones, metabolic, and antioxidant proteins, have been shown to be involved in the process of neuronal differentiation (ND) of neuroblastoma cell lines. No systematic approach to detect hitherto unknown and unnamed proteins or structures that have been predicted upon nucleic acid sequences in ND has been published so far. We therefore decided to screen hypothetical protein (HP) expression by protein profiling. Two‐dimensional gel electrophoresis with subsequent matrix‐assisted laser desorption/ionization–time of flight mass spectrometry (MALDI‐TOF/TOF) identification was used for expression analysis of undifferentiated and dimethylsulfoxide‐induced neuronally differentiated N1E‐115 cells. We unambiguously identified six HPs: Q8C520, Q99LF4, Q9CXS1, Q9DAF8, Q91WT0, and Q8C5G2. A prefoldin domain in Q91WT0, a t‐SNARE domain in Q9CXS1, and a bromodomain were observed in Q8C5G2. For the three remaining proteins, no putative function using Pfam, BLOCKS, PROSITE, PRINTS, InterPro, Superfamily, CoPS, and ExPASy could be assigned. While two proteins were present in both cell lines, Q9CXS1 was switched off (i.e., undetectably low) in differentiated cells only, and Q9DAF8, Q91WT0, and Q8C5G2 were switched on in differentiated cells exclusively. Herein, using a proteomic approach suitable for screening and identification of HP, we present HP structures that have been only predicted so far based upon nucleic acid sequences. The four differentially regulated HPs may play a putative role in the process of ND. Proteins 2006. © 2006 Wiley‐Liss, Inc.
Single Center Review of Femoral Arteriovenous Grafts for Hemodialysis
Authors: Englesbe Michael J. , Al‐Holou Wajd N. , Moyer Alice T. , Robbins Jessica , Pelletier Shawn J. , Magee John , Sung Randall S. , Campbell Darrell , Punch Jeffery D.
Published: January 2006

Introduction

It is unclear how to manage high risk hemodialysis patients who present with an indwelling catheter. The National Kidney Foundation Practice Guidelines urge prompt removal of the catheter, but the guidelines do not specifically address the problem of patients whose only option is a femoral arteriovenous (AV) graft.

Methods

This study was a retrospective review of all patients who underwent femoral AV graft placement for hemodialysis access between January 1, 1996 and January 1, 2003 at the University of Michigan Health System (UMHS). Graft patency is reported according to the standards developed by the Society of Vascular Surgery and the American Association of Vascular Surgeons.

Results

Thirty patients were identified who had undergone femoral AV graft placement. The mean follow‐up was 23 months (range 1–75 months). The patients had had significant medical co‐morbidities and multiple previous access operations (mean 3; interquartile range 1–5). The 1‐year secondary graft patency rate was 41%, the 2‐year rate was 26%, and the 3‐year rate was 21%. Infection was the cause of final graft loss in eight patients (50% of the grafts losses, 27% of the total grafts placed.) Among those who died (n = 14), the mean time from femoral graft placement to death was 31.2 ± 27.5 months. The patient survival was quite low: at 1 year 81%, at 2 years 68%, and at 3 years 54%.

Conclusions

These complex patients who have exhausted their upper extremity hemodialysis options do poorly following femoral AV graft placement. Consideration should be given to long‐term catheter‐based access in some of these patients.
Outcome and Treatment Strategy for Mid‐ and Lower‐Thoracic Esophageal Cancer Recurring Locally in the Lymph Nodes of the Neck
Authors: Motoyama Satoru , Kitamura Michihiko , Saito Reijiro , Maruyama Kiyotomi , Okuyama Manabu , Ogawa Jun‐ichi
Published: January 2006

The aim of the present study was to assess the outcome of treatment for patients with recurrent mid‐ and lower‐thoracic esophageal cancers in whom recurrence was localized to the lymph nodes of the neck, and to determine the best strategy for further treatment. Between 1989 and 2001, 270 patients with mid‐ and lower‐thoracic esophageal cancer underwent curative esophagectomy; 90 of those patients had a cancer recurrence. Our focus was on lymph node recurrence, especially when the recurrent cancers were localized to the lymph nodes in the neck. The outcomes of those patients and the efficacy of the strategies used to treat the recurrent cancers were determined. In 43 patients (48%), recurrent cancer initially appeared in the lymph nodes. Among the 43 patients, 15 (35%) had localized neck recurrence. The time between tumor recurrence and death among the 15 patients with localized neck recurrence was significantly longer than among the 28 patients with other recurrence patterns. In addition, 15 patients underwent lymph node resection, while 28 patients were treated non‐surgically. The time between tumor recurrence and death was significantly longer in patients treated surgically. Of the 15 patients in whom recurrence affected the neck lymph nodes only, 10 (67%) were treated surgically; their 2‐year survival rate after recurrence was 45%. The outcomes of recurrent esophageal cancers localized to the lymph nodes of the neck were better than those seen with other recurrence patterns, and salvage resection followed by chemoradiation therapy would seem to be indicated for those patients.
Typhoid Colonic Perforation in Childhood:A Ten‐Year Experience
Authors: Chang Yu‐Tang , Lin Jan‐You , Huang Yu‐Sheng
Published: January 2006

Background

Intestinal perforation resulting from a complication of typhoid fever has always been of concern because of its high morbidity and mortality rates. Most perforations occur in the terminal ileum.

Methods

Between May 1995 and June 2005, 24 children with typhoid colonic perforations receiving operations were retrospectively reviewed. The patients were 5 months to 86 months of age (mean: 25.1 ± 17.5 months). Surgical management consisted of primary closure of the perforation with ileostomy (70.8%), wedge resection and simple closure (20.8%), and partial colectomy with colostomy (4.2%). For the 11 patients with solitary cecal perforation, 6 received simple closure with ileostomy (group I), and wedge resection and simple closure was performed in the remaining 5 patients (group C). After operation, total parenteral nutrition (TPN) was instituted in each patient.

Results

Solitary cecal perforation is the most common type (45.8%). The length of hospital stay (LOS) varied from 12 days to 73 days, and the mean duration was 24.5 ± 14.7 days. The LOS showed no significant difference with regard to the location and the number of perforations. No mortality was noted in the entire series, but 19 patients had one or more complications, which may have prolonged the LOS; anemia and wound infections were the two most common problems. Group C had the shorter LOS (16.4 ± 3.4 days, versus 40.2 ± 22.6 days for group I) and fewer complications (0.6 ± 0.5, versus 3.25 ± 1.7 for group I). Any readmission for secondary operation to restore intestinal continuity in group C was therefore avoided.

Conclusions

To understand thoroughly the clinical course of typhoid fever is important. The typically high rate of complications after intestinal perforation secondary to typhoid fever may be reduced if operation is undertaken earlier. Solitary cecal perforation can be managed safely with wedge resection and simple closure without bowel exterioration. Routine usage of TPN may reduce the mortality associated with complications resulting from typhoid perforations.
Rapid Intraoperative Parathyroid Hormone Assay—More than Just a Comfort Measure
Authors: Hanif F. , Coffey J. C. , Romics L. , O‘Sullivan K. , Aftab F. , Redmond H. P.
Published: January 2006

Background

Minimally invasive radio‐guided parathyroidectomy (MIRP) has been embraced as an acceptable therapeutic approach to primary hyperparathyroidism. Preoperative sestamibi scanning has facilitated this technique. Here we evaluate the addition of a rapid intraoperative parathyroid hormone (iPTH) assay for patients undergoing MIRP.

Methods

A series of 51 patients underwent sestamibi localization of parathyroid glands followed by MIRP for primary hyperparathyroidism. Using peripheral venous samples, iPTH levels were measured prior to gland excision, as well as post‐excision at 5, 10, and 15 minutes, taking a 50% reduction in iPTH level as indicative of complete excision. Next, changes in serum iPTH were compared with preoperative and postoperative changes in serum calcium, as well as levels of intraoperative ex‐vivo radiation counts taken by hand‐held gamma probe.

Results

In this series, a drop of greater than 50% in iPTH levels was observed in 94% of patients (n = 48). Moreover, a significant drop in iPTH occurred within 10 minutes of excision in the majority (n = 42) of cases (P < 0.004). Changes in iPTH were comparable with the therapeutic reduction in calcium levels, as well as with the change in intraoperative ex‐vivo gamma counts.

Conclusions

This study demonstrates that the addition of an iPTH assay to MIRP provides a quick and reliable intraoperative diagnostic modality in confirming correct adenoma removal. Moreover, it precludes the requirement of frozen section.
Identification of Prognostic Factors Associated with Early Mortality after Surgical Resection for Pancreatic Cancer—Under‐analysis of Cumulative Survival Curve
Authors: Takamori Hiroshi , Hiraoka Takehisa , Kanemitsu Keiichiro , Tsuji Tatsuya , Hamada Chikuma , Baba Hideo
Published: January 2006

Background

The cumulative survival curve after surgery for advanced pancreatic cancer is characterized by a steep downward slope in the early postoperative period. The aim of this investigation was to identify the characteristics associated with early mortality in patients undergoing pancreatic resection for pancreatic cancer.

Methods

Thirty‐seven patients with extended radical pancreatectomy combined with intraoperative radiation therapy were studied. The cumulative survival curve in this series was depicted using the Kaplan‐Meier method. Assuming that there were two distinct survival curves, below and above the breakpoint, each part of the curve was modeled as an exponential distribution. Three parameters, the breakpoint, the high hazard rate below the breakpoint, and the low hazard rate above the breakpoint were estimated by the maximum likelihood method. Prognostic factors associated with early mortality after surgery were evaluated using univariate and multivariate Cox proportional hazards regression analyses.

Results

The breakpoint of the survival curve was estimated at 41 months. The short‐survival group (SSG) was defined as deceased earlier than 41 months after surgery, and included 31 patients (83.8 %). The long‐survival patient group (LSG) consisted of 6 patients who were alive more than 41 months after surgery. Eighteen SSG patients (58.1 %) died of hepatic metastases, whereas no LSG patients died of hepatic metastases. Abdominal pain and/or back pain during clinical course was identified by multivariate analysis as a prognostic factor for patients undergoing pancreatic resection.

Conclusions

The high hazard rate in the early postoperative period was closely linked with death due to liver metastases. The preoperative presence of local pain was a prognostic factor associated with early mortality.
Effects of Mosapride Citrate on Patients after Vagal Nerve Preserving Distal Gastrectomy Reconstructed by Interposition of a Jejunal J Pouch with a Jejunal Conduit for Early Gastric Cancer
Authors: Tomita Ryouichi , Ikeda Taro , Fujisaki Shigeru , Koshinaga Tugumichi , Tanjoh Katsuhisa
Published: January 2006

Background

Vagal nerve–preserving distal gastrectomy reconstructed by interposition of a jejunal J pouch with a jejunal conduit (hereinafter called DGP) is a function‐preserving operation for early gastric cancer. However, some patients after DGP have suffered from postprandial stasis in the substitute stomach, and postprandial stasis leads to abdominal symptoms. To clarify the significance of mosapride citrate (MS) for prevention of food stasis in the substitute stomach for patients after DGP, we studied the effects of MS before and after administration of MS.

Materials and Methods

In a total of 18 patients (10 men, 8 women; aged 34 to 70 years, average 63.1 years) during 5 years after DGP for early gastric cancer (Billroth I, D1 + α lymph node dissection, curability A), the relationship between their postoperative quality of life (QOL) and emptying function of the substitute stomach (EFS) was compared using a radioisotope method before MS therapy and after MS therapy at an oral dose of 15 mg/day for 3 months.

Results

(1) Interview. After MS therapy patients evidently had more appetite and ate more, with a slightly increase in body weight (0.5 ∼ 2 kg) compared with patients before MS therapy. Before and after MS therapy no patients had early dumping symptoms, and after MS therapy all patients clearly had fewer symptoms such as reflux esophagitis, nausea, and abdominal pain compared with before MS therapy. After MS therapy they also had significantly decreased abdominal fullness compared with before MS therapy (P = 0.0026). Endoscopically, we found reflux esophagitis in 2 patients from the before MS therapy group but in no patients from the after MS therapy group. All patients in the before MS therapy group showed residual contents in the substitute stomach, and seven patients in the after MS therapy group showed residual contents in the substitute stomach (P < 0.0001). There was a significant difference between before and after MS therapy (P < 0.0001). (2) EFS; The time to 50% residual rate of the before MS therapy group (80.5 ± 16.2 min) was significantly slower than that of the after MS therapy patients (65.6 ± 9.4 min) (P = 0.0091). After MS therapy (28.4% ± 5.2%), the residual rates at 120 minutes were significantly decreased compared with patients before MS therapy (38.2% ± 5.7%) (P = 0.0372).

Conclusions

Patients from the after MS therapy group clearly had improved gastric stasis compared with the before MS therapy group. These results showed more satisfactory QOL in patients after MS therapy. It is possible that MS therapy improves abdominal fullness due to the postprandial stasis in the substitute stomach, contributing to the improvement of QOL of patients after DGP.
Prevalence of pain in adults admitted to Catalonian hospitals: A cross‐sectional study
Authors: Vallano A. , Malouf J. , Payrulet P. , Baños J.E. ,   On behalf of the Catalan Research Group for Studying Pain in Hospital
Published: January 2006

Objective To survey the prevalence of pain in patients admitted to different hospitals of Catalonia and to describe which factors are related to pain.

Methods A cross‐sectional study was performed in 1675 patients from fifteen hospitals in Catalonia (Spain). Clinical and demographic data, as well as the existence of pain intensity evaluations and analgesic therapy, were obtained from medical charts. Characteristics of pain were given by patients after being interviewed by trained interviewers. The main‐outcome measure was the existence of pain (at the interview, in the previous 24 h, at the admission and at any time after admission) that was assessed by a visual analogue scale (VAS). The relationship of prevalence of pain to patients' characteristics was carried out by means of a multiple‐logistic‐regression model with pain presence as the dependent variable of interest.

Results A great variability in the prevalence and intensity of pain among different hospitals was observed. At the time of the interview, 48.5% (95% CI: 46.1–50.9%) of the patients had pain and the median VAS was 40 mm (range: 10–100 mm), and the prevalence of pain during the previous 24 h was similar (47.6%; 95% CI: 45.2–50%). At admission, 26.7% (95% CI: 24.6–28.8%) of patients were in pain, whereas 62% (95% CI: 59.7–64.3%) reported having pain at some time during their stay. Pain intensity annotations were absent in 51.3% (95% CI: 47.9–54.7%) of the medical records of the patients with pain. The factors associated with pain were younger age, female gender, presence of surgery, orthopaedic surgery wards, large hospital and prescribed analgesics.

Conclusion A high prevalence of clinically relevant pain in in‐patients was found as well as a great variability according to type of patients, clinical wards and hospitals. This study gives clear evidence of the lack of adequate management of pain in the majority of the hospitals and calls for the implementation of organisational and educational measurements that may settle this epidemic problem.
Reduction of human experimental muscle pain by alfentanil and morphine
Authors: Schulte Helène , Segerdahl Märta , Graven‐Nielsen Thomas , Grass Stefan
Published: January 2006

Musculoskeletal pain is a major clinical problem. By using various experimental models in humans, the understanding of the basic mechanisms behind muscle pain can increase, thereby giving hope for new and optimized treatment. Opioids are increasingly often used to treat muscle pain. There are, however, a limited number of previous studies on opioids and muscle pain, most of them using a relative low, single dose. Therefore, we wanted to further study the effect of two rather high doses of alfentanil (25 and 75 ng/ml) and morphine (0.14 and 0.28 mg/kg) in human volunteers. The study consisted of two parallel studies with morphine and alfentanil, respectively, and was conducted as randomized, double‐blinded, placebo‐controlled, 3‐way cross‐over. We used intramuscular infusion of hypertonic saline and intramuscular electrical stimulation to induce experimental pain. Visual analog scale (VAS)‐score, intramuscular electrical pain thresholds and pain area (local and referred) were measured.

Both alfentanil and morphine at their highest doses induced a 6 to 7‐fold increase in pain thresholds to single and repetitive (5 stimulations, 2 Hz) electrical stimulation. Alfentanil and morphine also reduced VAS score about 4 to 5‐fold during suprathreshold electric stimulation and during infusion of hypertonic saline. None of the drugs decreased referred pain. There were no apparent differences between the drugs, in terms of effect or adverse reactions.

In conclusion, this is the first study to compare two high doses of alfentanil and morphine on experimental muscle pain in humans. Both alfentanil and morphine reduced experimental muscle pain. There were no indications of any true pharmacodynamic differences between the two drugs.
The p53‐inducible E3 ubiquitin ligase p53RFP induces p53‐dependent apoptosis
Authors: Huang Jun , Xu Liang-Guo , Liu Ting , Zhai Zhonghe , Shu Hong-Bing
Published: January 2006

Recently, it has been shown that really interesting new gene (RING)–in between ring finger (IBR)–RING domain‐containing proteins, such as Parkin and Parc, are E3 ubiquitin ligases and are involved in regulation of apoptosis. In this report, we show that p53‐inducible RING‐finger protein (p53RFP), a p53‐inducible E3 ubiquitin ligase, induces p53‐dependent but caspase‐independent apoptosis. p53RFP contains an N‐terminal RING–IBR–RING domain and an uncharacterized, evolutionally highly conserved C‐terminal domain. p53RFP interacts with E2 ubiquitin‐conjugating enzymes UbcH7 and UbcH8 but not with UbcH5, and this interaction is mediated through the RING–IBR–RING domain of p53RFP. Interestingly, the conserved C‐terminal domain of p53RFP is required and sufficient for p53RFP‐mediated apoptosis, suggesting p53RFP‐mediated apoptosis does not require its E3 ubiquitin ligase activity. Together with a recent report showing that p53RFP is involved in ubiquitination and degradation of p21, a p53 downstream protein promoting growth arrest and antagonizing apoptosis, our findings suggest that p53RFP is involved in switching a cell from p53‐mediated growth arrest to apoptosis.
Differential mechanisms of glutamate receptor regulation of SynGAP in cortical neurones
Authors: Rockliffe Nichola , Gawler Debra
Published: January 2006

One prime candidate linkingN‐methyl‐d‐aspartate (NMDA) receptors to the regulation of the MAP kinase cascade is SynGAP, a negative regulator of Ras. In order to assess how a physiological stimulus can alter SynGAP activity, an appropriate whole cell system must be used and SynGAP must be specifically extracted from membranes whilst preserving the catalytic activity of the protein. Here, we have achieved this and studied the effect of NMDA/α‐amino‐3‐hydroxy‐5‐methylisoxazole‐4‐propionic acid (AMPA) and kainate receptor stimulations on SynGAP activity in cortical neurones. Furthermore, we have examined the role of extracellular Ca2+, CaM kinase II and the PSD‐95‐NR2B subunit interaction in SynGAP activity regulation and propose a novel convergence of signalling between AMPA, kainate and NMDA receptors.
BIOREL: The benchmark resource to estimate the relevance of the gene networks
Authors: Antonov Alexey V. , Mewes Hans W.
Published: January 2006

The progress of high‐throughput methodologies in functional genomics has lead to the development of statistical procedures to infer gene networks from various types of high‐throughput data. However, due to the lack of common standards, the biological significance of the results of the different studies is hard to compare. To overcome this problem we propose a benchmark procedure and have developed a web resource (BIOREL), which is useful for estimating the biological relevance of any genetic network by integrating different sources of biological information. The associations of each gene from the network are classified as biologically relevant or not. The proportion of genes in the network classified as “relevant” is used as the overall network relevance score. Employing synthetic data we demonstrated that such a score ranks the networks fairly in respect to the relevance level. Using BIOREL as the benchmark resource we compared the quality of experimental and theoretically predicted protein interaction data.
Cholesterol loading augments oxidative stress in macrophages
Authors: Hung Yao-Ching , Hong Meng-Yen , Huang G. Steve
Published: January 2006

To investigate the molecular consequence of loading free cholesterol into macrophages, we conducted a large‐scale gene expression study to analyze acetylated‐LDL‐laden foam cells (AFC) and oxidized‐LDL‐laden foam cells (OFC) induced from human THP‐1 cell lines. Cluster analysis was performed using 9600‐gene microarray datasets from time course experiment. AFC and OFC shared common expression profiles; however, there were sufficient differences between these two treatments that AFC and OFC appealed as two separate entities. We identified 80 commonly upregulated genes and 48 commonly downregulated genes in AFC and OFC. Functional annotation of the differentially expressed genes indicated that apoptosis, extracellular matrix, oxidative stress, and cell proliferation was deregulated. We also identified 87 differentially expressed genes unique for AFC and 31 genes for OFC. The uniquely expressed genes of AFC are associated with kinase activity, ATP binding activity, and transporter activity, while unique genes for OFC are associated with cell signaling and adhesion. To validate the hypothesis that oxidative stress is a common feature for AFC and OFC, we performed a cluster analysis employing the genes related to oxidative stress, but we were unable to distinguish AFC from OFC in this manner. We performed real‐time RT‐PCR and ELISA on foam cells to examine the transcripts and secreted protein of interleukin 1 beta (IL1β). IL1β was rapidly induced in foam cells, but for AFC both RNA level and protein level dropped immediately and was attenuated. To detect levels of reactive oxygen species in foam cells we conducted hydroethidine staining and observed high levels of superoxide anion. We conclude that loading free cholesterol induces high levels of superoxide anion, increases oxidative stress, and triggers a transient inflammatory response in macrophages.
Salting‐in the microbial cytoplasm
Authors: Braun F.N.
Published: January 2006

Microbial organisms are known to rely for osmotic regulatory purposes on an assortment of low molecular weight molecules earmarked for function as osmolytes. The so‐called ‘compatible’ subclass of osmolyte, notably glycine betaine, is distinguished by a propensity to avoid the large bound fraction of cytoplasmic water adsorbed at the surface of biological macromolecules. Here we argue that this property is implicated in thermodynamic stabilisation of the cytoplasm. A rudimentary molecular statistical approach indicates that flooding the cytoplasm with large amounts of compatible osmolyte is an effective way to deal with the threat of phase separation.
Activation of p53 as a causal step for atherosclerosis induced by polycyclic aromatic hydrocarbons
Authors: Iwano Shunsuke , Shibahara Norihito , Saito Tetsuya , Kamataki Tetsuya
Published: January 2006

This study was performed to prove our hypothesis that the metabolite(s) of polycyclic aromatic hydrocarbons (PAHs) caused the activation or phosphorylation of p53 via DNA damage to suppress the liver X receptor (LXR)‐mediated signal transductions as a probably more direct mechanism. We found that LXR‐mediated trans‐activation was inhibited by 3‐methylchoranthrene (MC) and doxorubicin (Dox) in HepG2 cells carrying wild‐type p53, but not in Hep3B cells possessing mutant p53. The exogenous expression of wild‐type p53 suppressed the LXR‐mediated trans‐activation in Hep3B cells. The expression of mRNA for ATP binding cassette A1 was suppressed by MC and Dox in HepG2 cells. The protein expression of retinoid X receptor (RXR), a partner of LXR to form a heterodimer, was suppressed by MC and Dox in HepG2 cells.
Effects of water quality and stocking density on growth performance of juvenile cod (Gadus morhua L.)
Authors: Björnsson Björn , Ólafsdóttir Sólveig R.
Published: January 2006

In a 5-month experiment where groups of juvenile cod were reared in a flow-through system at low density (Group 1) and in a recirculating system at low (Group 2) and high densities (Group 3), the recirculated water had negative effects on growth rate, nutritional condition, and mortality. After the first month, mean weight was significantly larger in Group 1 than in the other two groups. The effect of stocking density on mean weight was not significant until the end of the experiment, when densities were 9.0 and 48.3 kg m−3 in Groups 2 and 3, respectively. Initial mean weight of fish was 37.3 g and at the termination of the experiment mean weights were 225.2, 181.8, and 167.9 g in Groups 1, 2, and 3, respectively. After 5 months, mean condition factors were 1.074, 0.965, and 0.946, mean liver indices 9.5, 9.0, and 7.6, and mean mortalities over the course of the experiment were 1.0%, 5.1%, and 2.4% in Groups 1, 2, and 3, respectively. Water temperature (10.4–10.6°C), pH (7.2–7.4) and oxygen concentration (8–10 mg l−1) were similar among groups. Concentration of total ammonia nitrogen (TAN) was similar among groups for the first half of the experiment (0.3–0.4), but during the last month it was 0.6, 1.3, and 1.5 mg l−1 in Groups 1, 2, and 3, respectively. There was a negative correlation (r2 = 0.48, n = 36) between relative growth rate and TAN, suggesting that ammonia may have been a limiting factor in the recirculating system. The apparent threshold limit of TAN for reduced growth was approximately 1 mg l−1.