bluuebunny/crossref_metadata_2025 huggingface dataset:
1141160720 Does the anti‐proliferative action of ovine uterine serpin involve apoptosis?
Authors: Padua MB , Hansen PJ
Published: May 2006

Ovine uterine serpin (OvUS) is the major progesterone‐induced protein in the uterus of the pregnant sheep. This protein is a member of the serine proteinase inhibitor superfamily and it has been proposed to down‐regulate uterine immune function during pregnancy to protect the fetus. In vitro experiments have shown that both the native and the recombinant (r) form of the protein can inhibit mitogen‐induced lymphocyte proliferation and growth of canine primary osteogenic sarcoma cells, mouse lymphoma cells, human prostatic adenocarcinoma cells (PC‐3 cell line) and bovine preimplantation embryos. The mechanism by which OvUS inhibits cell proliferation is still unknown. Accordingly, experiments were conducted to test whether rOvUS exerts its anti‐proliferative action through apoptosis. In the first experiment, the anti‐proliferative effect of rOvUS on PC‐3 cells was tested to determine the minimal concentration effective at inhibiting proliferation. Proliferation was inhibited by concentrations of OvUS at 8 μg/mL and higher. The incorporation of [3H]thymidine was 4043, 3998, 3464, 2785, 2827, 2310, and 2332 dpm for 0, 0.5, 2, 8, 32, 125, and 250 μg/mL, respectively. In the second experiment, PC‐3 cells were cultured for 48 hr with 50, 100 and 200 μg/mL of rOvUS or ovalbumin. Cells were then fixed and apoptotic cells identified using the TUNEL assay to detect DNA fragmentation. There was no effect of OvUS at any concentration tested on the percent of cells that were apoptotic. Percent apoptosis was 1.3% for control cells, 3.8%, 4.8% and 2% for cells cultured with 50, 100 and 200 μg/mL rOvUS, and 2% for cells cultured with 200 μg/mL ovalbumin. Results confirm that OvUS inhibits proliferation of PC‐3 cells and indicate that inhibition does not involve induction of apoptosis. Further experiments are warranted to elucidate the anti‐proliferative mechanism of action of OvUS.
1141170685 Tumor‐infiltrating lymphocytes contain higher proportion of FOXP3+ T lymphocytes from cervical cancer than that from cervical intraepithelial zneoplasm
Authors: Kuo TY , Ho HN
Published: May 2006

Cervical cancer (CC) is preceded by well‐defined changes in the epithelium known as cervical intraepithelial neoplasm (CIN). Our previous studies have revealed that the subpopulations and functions of tumor‐ infiltrating lymphocytes (TIL) from CC are altered. Dysfunction of the host immune system in cancer patients can be due to a number of reasons including the inhibitory functions of regulatory T (Treg) cells. FOXP3 has been shown to be a master control gene for the development and function of CD4+ CD25+ Treg cells. To characterize the functional role of Treg cells in progress of CC, we compared samples from four groups: CIN 2, CIN3, CC with and without tumor draining lymph node. By using the immunofluorescence staining and confocal‐based image quantitative analysis in paraffin‐embedded tissue sections, excess in the presence of FOXP3+ cells is observed from CC compared to that from CIN2/3. The significant difference of FOXP3+ cells expression level between CIN and CC indicate that the Treg might play an important role in the progression of CC.
1141180267 Mechanism of HIV transmission across female primary genital epithelial cells
Authors: Fernandez SA , Bunce M , Kaushic C
Published: May 2006

Our lab is utilizing human primary genital epithelial cultures to investigate the mechanism of HIV transmission across the female genital mucosa. Primary endometrial epithelial cells (ECs) grown on matrigel‐ coated cell inserts were used for HIV infection studies. EC's were infected with cell‐ free or cell‐ associated R5 and X4 virus strains. Infections were performed in the presence or absence of the appropriate macrophage (U937) or T‐cell (Jurkat) target cell‐ line in basolateral compartments of cultures. Virus was quantified from supernatants using p24 ELISA. U373‐CXCR4 and CCR5 magi cell lines were utilized to measure infectious virus and tropism. Sybrgreen real time PCR was used to detect viral RNA in cell lysates. Infectious HIV particles were found only in basolateral supernatants of EC's infected with X4 strains of HIV in the presence of appropriate target cells. This indicated that EC's may not be productively infected, but were only able to transmit HIV in the presence of target cells. These results were supported by the detection of higher levels of gag gene (CT‐15) in X4 target cells compared to EC's (CT‐25) using real‐time PCR. No infectious virus was present in primary EC's infected with cell‐free R5 strain, although gag gene product was detected by real time PCR in both R5 target cells as well as EC's. P24 levels in supernatants were not indicative of infectious virus. Our data indicates that R5 and X4 virus strains have differential abilities to cross the female genital mucosa to infect target cells. The presence of target cells appears to be critical for the production of infectious HIV particles under these culture conditions. These studies are providing important information regarding the ability of HIV‐1 to cross the female genital mucosa.
1141180772 Activated NK cells in spontaneous abortion cases induce apoptosis on extravillous trophoblast in a granulysin dependent manner
Authors: Nakashima A , Shizaki A , Myojo S , Saito S
Published: May 2006

Extravillous trophoblast (EVT) is well known to undergo apoptosis in spontaneous abortion cases although the apoptotic mechanism is still unclear. Granulysin (Gr) is a cytotoxic granule protein of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) with tumoricidal activities. We have reported that the expression of granulysin, which was mainly produced by decidual NK cells, is significantly higher in the deciduas of spontaneous abortion cases than early normal pregnancy. We then have attempted to investigate the molecular mechanism of apoptosis associated with Gr. We also assessed apoptosis in the HTR8/SV40 cell, extravillous trophoblast (EVT) cell line, which were transfected with GFP‐fused Gr. Anti‐cleaved cytokeratin 18 antibody and Annexin‐V were used for detecting apoptosis by IS and FC. We detected the apoptosis of the EVT in miscarriage cases. In those EVT, we detected nuclear staining of Gr. We next examined the effect of Gr with the target trophoblast cells by using GFP‐fused Gr vector. Exogenously expressed GFP‐fused Gr preferentially accumulated in nucleus from cytoplasm, resulting in apoptosis. Similar findings were obtained by native granulysin produced by IL‐2 stimulated decidual lymphocytes. Our findings suggest that the granulysin‐positive NK cells might attack EVT and they release granulysin into the intracellular space, resulting in apoptosis of EVT. So far, other groups have reported that Fas‐FasL pathway and and/or TNF‐α pathway is associated with apoptosis of EVT. In additiont, we propose granulysin‐positive NK cells‐induced apoptosis as a new mechanism for EVT cell death.
1141222326 Analysis of effects of androgen on protein profiles in TM4 mouse Sertoli cells by SELDI‐TOF mass spectrometry
Authors: Komori S , Sakata K , Kasumi H , Takenobu T , Uchida K , Fukuoka M , Koyama K
Published: May 2006

Aim:  The identification of proteins induced by androgen in Sertoli cells is a major issue in spermatogenesis.

Methods:  In this study, we analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone using SELDI‐TOF mass spectrometry.

Results:  We found increases in expression of a 5.0 kDa protein at 15 min, a 11.3 kDa protein at 24 hr and 4.3, 5.7, 5.8, 9.95 and 9.98 kDa proteins at 48 hr. On the other hand, expression of 6.3 and 8.6 kDa proteins were decreased at 30 min, and 4.9, 5.0, 12.4 and 19.8 kDa proteins at 48 hr. The 11.3 kDa molecule was identified as macrophage migration inhibitory factor (MIF) known to having various functions. The 9.98 kDa molecule was identified as calgizzarin related to calcium channel. The timing of its expression suggests that MIF and calgizzarin are involved late in androgen regulation of spermatogenesis in Sertoli cells. The 19.8 kDa molecule was identified as translationally controlled tumor protein (TCTP) known to bind to tublin and Ca2+ and regulates cell proliferation.

Conclusion:  Androgenic effects on spermatogenesis are exerted eventually through MIF, calgizzarin and TCTP.
1141424444 Detection of a2V‐ATPase in T regulatory cells of women with recurrent spontaneous abortions or implantation failures
Authors: Ntrivalas EI , Fukui A , Gilman‐Sachs A , Levine R , Kwak‐Kim J , Beaman KD
Published: May 2006

Problem:  T regulatory cells (Tregs) have recently been shown to play a critical role in maternal tolerance to the fetus. Tregs are decreased in women with recurrent miscarriages. a2V‐ATPase (previously referred to as Regeneration and Tolerance Factor) is expressed in activated lymphocytes and plays a role in immune regulation. The aim of this study was to investigate the expression of a2V‐ATPase on CD4+/CD25bright Tregs.

Method of Study:  Whole blood from women with RSA or implantation failures was reacted with anti‐CD4 and anti‐CD25 mAbs for the identification of CD4+/CD25bright and CD4+/CD25neg T cells by flow cytometric analysis. Subsequently, these two T‐cell populations were analyzed for the expression of a2V‐ATPase using PE‐conjugated 2C1 mAb (specific for the membrane portion of a2V‐ATPase). These two cell populations were also analyzed for the expression of CD71, CD62L, CD45RO and CD58 (Treg markers).

Results:  a2V‐ATPase was more highly expressed on CD4+/CD25bright Tregs (22.8 ± 16.4%) than on CD4+/CD25neg T cells (2.4 ± 3.8%) in women with RSA (P < 0.0001). Additionally, a2V‐ATPase was more highly expressed on CD4+/CD25bright Tregs (18.0 ± 18.2%) than on CD4+/CD25neg T cells (1.5 ± 1.4%) in women with implantation failures (P < 0.0001). a2V‐ATPase expression also coincided with the expression of CD71, CD62L, CD45RO and CD58 in Tregs, as opposed to the conventional CD4+/CD25neg T cells.

Conclusions:  The expression of a2V‐ATPase in Tregs of women with RSA or implantation failures is a novel finding and suggests that this vacuolar ATPase plays an important role in suppression. a2V‐ATPase may be a unique molecule in the identification of Tregs among peripheral blood lymphocytes and may also explain the tolerogenic activity of these cells.
1141612296 Combined treatment of GnRH analog plus aromatase inhibitor versus GnRH analog
Authors: Scarpellini L , Scarpellini F
Published: May 2006

Uterine leiomyomatosis occurs frequently in the women population. Pharmacological treatment are suitable for this pathology, such as GnRH analogs. With these substances it has been achieved a dramatic shrinkage of leiomyomatas and uterus with a consequent rapid recovery from associated complications. However, long period of treatment, and consequent prolongated estrogen deprivation may give several problems on general physical conditions of women. In order to reduce the problems due to the lack of estrogen activity during GnRH treatment, reducing the therapy time, we add a low dose of aminogluthetimide, an aromatase inhibitor substance. A controlled study was conducted in leiomyomatosis women, to compare GnRH analog plus aminogluthetimide versus GnRH analog alone. 67 consecutive menstruated women suffering of leiomyomatosis and menometrorrhagia, referred to outpatient clinic of Ob/Gyn Department, ‘La Sapienza’ University, Roma, Italy, were enrolled in the study. Patients were randomly assigned to one of the two arms of the study. 33 patients were treated with 3.75 mg subcutaneous Goserelin every 28 days and 250 mg/daily of aminogluthetimide (group A). Thirty‐four women were treated with 3.75 mg subcutaneous Goserelin every 28 days alone (group B). Serum estradiol concentration and ultrasound uterine dimension were evaluated 1 day before the beginning of therapy, after 2 months and at the end of treatment. The two groups of patients were similar for age, body mass index, serum estradiol levels and uterine volume before treatment. After 2 months of treatment in group A serum estradiol levels were 26.9 ± 3.0 pg/mL and in group B 29.4 ± 23.4 pg/mL. Uterine volume were 318 ± 43 cm2 for group A (reduction of 23.5 ± 7.9%) 369 ± 49 cm2 for group B (reduction of 12.3 ± 6.1%) (P < 0.05). The duration of the treatment in group A was 4.3 ± 0.4 months whereas in group B was 6.2 ± 0.6 months (P < 0.01). At end of treatment in group A serum estradiol levels were 21.5 ± 2.7 pg/mL and in group B 26.4 ± 3.0 pg/mL. Uterine volume were 197 ± 61 cm2 for group A (reduction of 52.6 ± 10.3%) 243 ± 51 cm2 for group B (reduction of 42.2 ± 11.4%) (P < 0.05). Combined treatment of GnRH analog and aromatase inhibitor was more effective in reducing uterine volume than GnRH analog treatment alone, also in term of time. Since serum estradiol concentration in the two group did not show significative differences, the more rapid and dramatic effect observed in combined treatment may be due to an inhibition of aromatase enzyme in the leiomyomata tissue. The combined treatment allows a sensible reduction of treatment time, reduction of hypoestrogenic period and lower costs.
1141635071 Suppressor of Cytokine Signalling 3 (SOCS3) controls trophoblast invasion and proliferation
Authors: Poehlmann TG , Markert UR
Published: May 2006

Introduction:  Signal Transducer and Activator of Transcription 3 (STAT3) is an intracellular mediator of trophoblast invasion. In other cells, Suppressor Of Cytokine Signalling 3 (SOCS3) is known to be a major antagonist of STAT3 activation. For auto‐regulation and to avoid excessive signals, SOCS3 gene expression is partly induced by STAT3.

Methods:  The human choriocarcinoma cell line JEG‐3 was used for all experiments. STAT3 expression and STAT3 tyrosine (705) phosphorylation were analyzed by polyacrylamid gele electrophoresis followed by Western blotting. Interleukin‐6 (IL‐6) or Leukemia Inhibitory Factor (LIF) were used for stimulation. SOCS3 expression was silenced by RNA interference. For this approach in a first step, small interfering RNA (siRNA) was used. Subsequently, Jeg‐3 cells were transfected with genes for the respective, self‐designed small hairpin RNA (shRNA), neomycin resistance and green fluorescent protein. Application of neomycin and cell sorting by green fluorescence were used for elimination of non‐transfected cells. Non‐genomic siRNA and shRNA were respectively used for controls. Colorimetric proliferation assays were performed.

Results:  In IL‐6 stimulated cells, SOCS3 knockdown leads to increased STAT3 tyrosine phosphorylation. After LIF stimulation, proliferation in SOCS3 knockdown cells is significantly higher than in controls. STAT3 expression is not affected by SOCS3 knockdown.

Conclusion:  SOCS3 plays a major role in intracellular regulation of trophoblast proliferation and invasiveness. The balance of STAT3 and SOCS3 may tune the behavior of invading trophoblast cells.
1141636674 Differential serine and tyrosine phosphorylation of Signal Transducer and Activator of Transcription 3 (STAT3) in Jeg‐3 choriocarcinoma cell lines
Authors: Roediger J , Busch S , Enkelmann A , Poehlmann TG , Markert UR
Published: May 2006

Background:  Signal Transducer and Activator of Transcription 3 (STAT3) is an intracellular signalling molecule, which is used by several cytokines, including leukemia inhibitory factor (LIF), epithelial growth factor (EGF), and interleukin‐6 (IL‐6). It induces a variety of gene transcripts and cell functions. In trophoblast cells and in tumor cells, its tyrosine phosphorylation is directly linked to their invasiveness. The regulation and function of STAT3 serine phosphorylation is still widely unclear.

Material and Methods:  Jeg‐3 choriocarcinoma cells were stimulated with different concentrations of EGF, IL‐6 and LIF. STAT3 serine (727) and tyrosine (705) phosphorylation were analyzed 5–60 min after stimulation by SDS‐PAGE electrophoresis followed by Western blotting.

Results:  Jeg‐3 cells display spontaneous STAT3 serine phosphorylation. 100 ng/mL EGF induces a time‐dependent reduction starting 15 min after stimulation. Tyrosine phosphorylation does not occur spontaneously, but is strongly induced by EGF at all analyzed time points. LIF induces tyrosine phosphorylation, but affects serine phosphorylation only very slightly. IL‐6 did not influence neither serine phosphorylation nor tyrosine phosphorylation.

Discussion:  The EGF induced STAT3 tyrosine phosphorylation may be responsible for its invasion triggering capacities. The parallel reduction of serine phosphorylation may enhance this effect. LIF was formerly shown to enhance trophoblast invasion via STAT3 tyrosine phosphorylation. IL‐6 displays very little effects on STAT3 and seems to use other pathways for signalling.
1141638491 Hepatocyte growth factor (HGF) stimulates mammalian target of rapamycin (mTOR) in choriocarcinoma cell lines and human trophoblast cells
Authors: Busch S , Poehlmann TG , Roediger J , Markert UR
Published: May 2006

Introduction:  Hepatocyte growth factor (HGF), interleukin‐6 (IL‐6) and insulin‐like growth factor‐II (IGF‐II) are involved in the regulation of trophoblast cell migration and invasion. Signal Transducer and Activator of Transcription 3 (STAT3) and Mammalian Target Of Rapamycin (mTOR) signalling regulate cell invasion, growth and proliferation. mTOR plays also a key role during embryogenesis. Knock‐out mice embryos die after implantation and blastocysts trophoblast outgrowth is reduced.

Aim:  Stimuli which might trigger such invasive behaviour through mTOR should be defined.

Methods:  The human choriocarcinoma cell lines JEG‐3, JAR, the human choriocarcinoma‐trophoblast hybrid AC1‐M59 and human term trophoblast cells were stimulated with HGF, IL‐6 or IGF‐II. At several time points, the phosphorylation level of mTOR and STAT3 were tested by Western blot. STAT3 DNA‐binding capacity was analyzed by Electrophorectic Mobility Shift Assay (EMSA). To examine the role of mTOR for invasion and proliferation, mTOR expression was silenced by RNA interference (RNAi).

Results:  HGF, IGF‐II and IL‐6 did neither induce tyrosine (705) phosphorylation of STAT3 nor STAT3 DNA binding capacity as assessed by EMSA. HGF led to an increase of mTOR serine (2448) phosphorylation for all cell types after 15 and 30 min while IL‐6 and IGF‐II did not induce mTOR phosphorylation. Simultaneously, HGF decreased STAT3 serine (727) phosphorylation. mTOR silencing in AC1‐M59 correlates with reduced proliferation and invasion. STAT3 expression was not affected by mTOR knock down.

Conclusion:  HGF triggers mTOR activity in trophoblast and trophoblast‐like cells. mTOR is a main regulator of crucial trophoblast functions.
1141646904 Influence of conjugated linoleic acid (CLA) on proliferation and cytokine synthesis in maternal, fetal and decidual lymphocytes after stimulation with allergens
Authors: Enke U , Heinzelmann J , Markert UR , Jahreis G , Seyfarth L
Published: May 2006

Background:  Fetal programming is the notion that adverse environmental conditions in utero can cause short term survival adaptations that may have long‐term consequences, such as chronic disease in subsequent lifetime. Recently, some authors reported that the increase of allergy prevalence in childhood may be linked with fetal immune development. In this regard, literature survey inspired to study the influence of two conjugated linoleic acid (CLA)‐isomers (c9,t11 and t10,c12) on parameters of the immune system in pregnancy.

Methods:  Lymphocytes from allergic and non‐allergic mothers, cord blood of their newborns, and the decidual layer of term placentae were isolated and cultured for 2 days and supplemented or not with relevant allergens, c9,t11 CLA, t10,c12 CLA or linoleic acid, respectively. Expression of CD69 (activation marker) and CD71 (transferrin receptor; proliferation marker) on B and T lymphocytes and T helper cell type 1 (Th1) and Th2 cytokines in culture supernatant were analyzed.

Results:  Both CLA isomers led to an increase of the IFN‐γ/IL‐10 ratio in supernatants of peripheral maternal and cord blood cells from allergic patients and non‐allergic‐control. CLA supplementation decreased IL‐10 secretion of peripheral maternal and decidual lymphocytes and the portion of CD71+ maternal B cells. Linoleic acid induced a similar effect.

Conclusion:  An immunological effect of CLA on maternal and fetal lymphocyte responses could be demonstrated. It would be expected that further investigations could reveal differences in effects of CLA and linoleic acid.
1142280946 Effect of estradiol and PAMPs on class II mediated antigen presentation and immunomodulatory molecule expression in the mouse female reproductive tract
Authors: Wira CR , Rossoll RM , Ochiel DO , Haddad SN , Schaefer TM
Published: May 2006

Problem:  Antigen presenting cells (APC) in the female reproductive tract play important roles in innate immune defense and activation of the adaptive immune responses. The objective of this study was to examine the effects of estradiol and PAMP on antigen presentation in the female reproductive tract.

Method of Study:  DO11.10 T cell antigen receptor transgenic mice specific for the MHC class II‐restricted OVA323–339peptide were used to study the effects of estradiol and PAMP on antigen presentation of OVA by uterine epithelial (EC) and stromal cells as well as vaginal cells to OVA specific memory‐T cells.

Results:  Estradiol inhibited antigen presentation of OVA by uterine EC, uterine stromal cells and vaginal cells to OVA specific memory‐T cells. When ovariectomized animals were treated with estradiol for 1 or 3 days, antigen presentation decreased by 20–80%. In contrast, incubation with TLR agonists increased antigen presentation by EC (Poly (I:C), Pam3Cys), stromal cells (PGN, Pam3Cys) and vaginal cells (LPS, Pam3Cys). Analysis of mRNA expression by real time RT‐PCR indicated that estradiol inhibited CD40, CD80/86 and class II in the uterus and vagina. In contrast, stimulation of antigen presentation by PAMP did not correlate with changes in costimulatory molecule mRNA expression.

Conclusions:  These results indicate that APC in the uterus and vagina are responsive to estradiol, which inhibits antigen presentation and costimulatory molecule expression. These findings suggest that whereas APC in the uterus and vagina respond to TLR agonists with increased antigen presentation, which initiates an adaptive immune response, their effects appear to be at levels other than the expression of costimulatory molecules.

Acknowledgement:  Supported by AI‐13541 from NIH.
1142292808 Developmental exposure of mice to dioxin exhibit endometrial dysfunction which mimics endometrium from women with endometriosis
Authors: Bruner‐Tran Kaylon L. , Nayyar Tultul , Piestrzeniewicz‐Ulanska Dagmara , Eisenberg Esther , Osteen Kevin G.
Published: May 2006

Problem:  The environmental contaminant 2,3,7,8‐tetracholorodibenzo‐p‐dioxin (TCDD) has been suggested to play a role in the etiology of endometriosis, but whether women develop this disease due to a prior exposure to endocrine disruptors is difficult to ascertain. Several studies have failed to demonstrate a link between adult TCDD exposure and the development of endometriosis, but we suspect it is the very early exposures that may be most relevant to alterations in adult tissue function. To this end, we have recently developed a mouse model in which we can examine the impact of developmental TCDD exposure on the adult uterus. The endometrial phenotype of these mice is markedly similarly to the phenotype found in women with endometriosis.

Method of Study:  TCDD was administered to mice at multiple developmental stages (in utero, prepubertal and pubertal). Adult mice were ovariectomized and treated with either estradiol (E2) or with E2 and progesterone (P4). The uteri were collected and PR expression examined by immunohistochemistry. As an indicator of P4 responsiveness, tissues were further examined for expression of TGF‐β2. Human endometrial samples from women with and without endometriosis were similarly examined for PR and TGF‐β2 expression.

Results:  Minimal uterine expression of PR and TGF‐β2 was observed in TCDD exposed mice while control mice demonstrated abundant expression of both proteins. Analysis of normal human endometrium revealed intense staining for both PR and TGF‐β2 during the P4‐dominated secretory phase while these proteins were greatly diminished in tissues from women with endometriosis.

Conclusions:  A reduced responsiveness to P4 has recently been demonstrated in women with endometriosis. Although development of endometriosis is likely multifactorial, our studies are consistent with a role of in utero/developmental exposure to TCDD in the pathophysiology of this disease.

Acknowledgement:  This work was supported by NIEHS # R21ES12298 and The Endometriosis Association.
1141099968 Immunization under the influence of estradiol leads to non‐sterile protection against genital herpes infection
Authors: Gillgrass AE , Chege D , Kaushic C
Published: May 2006

Hormones play an important role in the induction of protective immune responses after intravaginal (IVAG) immunization with an attenuated strain of herpes simplex type 2 (HSV‐2 TK‐). Previously we reported that ovariectomized (OVX) mice receiving estradiol (E) prior to immunization were not protected, while mice receiving progesterone (P) had sterile protection after IVAG HSV‐2 challenge. Protection correlated with the ability of TK‐ to cause productive infection. Lack of protection in the E group was associated with the effect of E on thickening of vaginal epithelium. In the current study, we examined the effect of E on immunization after the thickening of vaginal epithelium subsided. OVX, C57BL/6 mice were given P, E or saline (S) for 3 days. The P and S groups were then immunized with IVAG TK‐. To determine the time point at which E mice could be successfully immunized, TK‐ was given between days 1 and 7 (E1–E7) post‐E treatment. Three weeks after immunization, mice were challenged with IVAG HSV‐2. Protection was absent in E1–E4 groups, 100% in E6 and E7 and partially successful in E5. While the protection in P and S was sterile, protected mice in E5 and E6 groups had non‐sterile protection. The presence of HSV‐2 specific IgG antibodies in the vaginal washes after challenge closely correlated with sterile immunity. After challenge, vaginal associated lymphoid tissues (iVALTs) were seen in the vaginal tract of the protected groups, except for the E5 group. The cytokine environment after challenge was also examined. The results show that while immunization under the effect of E may lead to clearance of virus, the mechanism is quite different than that seen under other hormonal conditions. Local hormonal influences must be considered when designing vaccines for STDs.
Middle segment pancreatectomy for the treatment of benign and low malignant potential pancreatic tumours
Authors: Lou Wen‐Hui , Jin Da‐Yong , Niu Wei‐Xin , Ni Xiao‐Lin , Wu Wen‐Chaun
Published: May 2006

Background:  Conventional distal pancreatectomy is used for the removal of lesions located at the body and tail of the pancreas. Generally, the spleen is sacrificed. But for benign and low malignant potential tumours in the pancreatic neck and body, this incurs notable loss of normal pancreatic tissue and the unnecessary risk of diabetes mellitus as well as splenic loss.

Methods:  We report three cases of middle segment pancreatectomy in an effort to avoid the unnecessary loss of normal pancreatic tissue and the spleen. Two patients were males with a mucinous cystadenoma and non‐cystic mucinous cystadenocarcinoma. The other was a female with a serous cystadenoma. Middle segment pancreatectomy was performed. Two patients were reconstructed with double pancreaticojejunostomy and the third with a pancreaticogastrostomy.

Results:  No major complications were observed.

Comnclusions:  Middle segment pancreatectomy is a safe and effective procedure for the resection of benign and low malignant potential tumours in the pancreatic neck and body with preservation of normal pancreatic tissue and the spleen in selected patients. This is the first report of middle segment pancreatectomy for pancreatic tumour in China.
Postoperative complications of laparoscopic adjustable gastric banding
Authors: Dukhno Oleg , Pinsk Ilya , Levy Isaac , Ovnat Amnon
Published: May 2006

Background:  The primary aim was to determine the rate and nature of late postoperative complications in relation to the type of band used and the route through which it encircled the stomach in a retrospective review of postoperative complications that necessitated reoperation in 500 consecutive cases of laparoscopic adjustable gastric banding (LAGB) conducted over the past 48 months.

Methods:  The four study groups were: (A) Lap‐Band® perigastric route, 186 cases; (B) Lap‐Band® pars flaccida route 73 cases; (C) SAGB perigastric 70 cases, and (D) SAGB pars flaccida, 171 cases. There were 414 women and 86 men. The mean age was 39.2 ± 11.0 years (17–68). The mean weight was 126 kg, and mean BMI was 43.4 kg/m2 prior to operation.

Results:  There were 32 cases of slippage (17%) in group A and five cases (7%) in Group B (P < 0.05). In group C there were 15 cases (21%) of slippage and in group D six cases (3%) (P < 0.05). There were no significant differences between Groups A and C or Groups B and D. Except for one case of band erosion in Group C, erosion, tube disconnection and aneurysmatic balloon dilatation occurred only in groups A and B. Band erosion occurred in 2% each in groups A and B, tube disconnection in 3% and 4%, respectively, and balloon dilatation in 2% in each of groups A and B.

Conclusions:  Slippage was primarily route dependent. Erosion, tube disconnection and aneurysmatic dilatation of the balloon were encountered almost exclusively in the high‐pressure system. We conclude that the pars flaccida route with the low‐pressure system is more effective in reducing the rate of complications necessitating reoperation.
Transperitoneal laparoscopic radical prostatectomy: The Tuen Mun Hospital experience
Authors: Chan Steve Wai‐Hee
Published: May 2006

Background:   The technique of laparoscopic radical prostatectomy was popularized after the report by a French group in 1998. The Tuen Mun Hospital started the program of laparoscopic radical prostatectomy since 2002 and the surgical technique has been evolving along a learning curve.

Methods and results:   We reviewed the transperitoneal approach of this procedure with emphasis on the variation of technique that the Tuen Mun Hospital has adopted.

Conclusion:   Although well described and standardized, the technique of transperitoneal laparoscopic radical prostatectomy is still evolving, as are many other surgical procedures. The exposure and development of different techniques allows the surgeons to be better equipped in catering for patients with different needs.
A Methodology for Simulating Compressible Turbulent Flows
Authors: Fasel Hermann F. , von Terzi Dominic A. , Sandberg Richard D.
Published: May 2006

A flow simulation Methodology (FSM) is presented for computing the time-dependent behavior of complex compressible turbulent flows. The development of FSM was initiated in close collaboration with C. Speziale (then at Boston University). The objective of FSM is to provide the proper amount of turbulence modeling for the unresolved scales while directly computing the largest scales. The strategy is implemented by using state-of-the-art turbulence models (as developed for Reynolds averaged Navier-Stokes (RANS)) and scaling of the model terms with a “contribution function.” The contribution function is dependent on the local and instantaneous “physical” resolution in the computation. This physical resolution is determined during the actual simulation by comparing the size of the smallest relevant scales to the local grid size used in the computation. The contribution function is designed such that it provides no modeling if the computation is locally well resolved so that it approaches direct numerical simulations (DNS) in the fine-grid limit and such that it provides modeling of all scales in the coarse-grid limit and thus approaches a RANS calculation. In between these resolution limits, the contribution function adjusts the necessary modeling for the unresolved scales while the larger (resolved) scales are computed as in large eddy simulation (LES). However, FSM is distinctly different from LES in that it allows for a consistent transition between RANS, LES, and DNS within the same simulation depending on the local flow behavior and “physical” resolution. As a consequence, FSM should require considerably fewer grid points for a given calculation than would be necessary for a LES. This conjecture is substantiated by employing FSM to calculate the flow over a backward-facing step and a plane wake behind a bluff body, both at low Mach number, and supersonic axisymmetric wakes. These examples were chosen such that they expose, on the one hand, the inherent difficulties of simulating (physically) complex flows, and, on the other hand, demonstrate the potential of the FSM approach for simulations of turbulent compressible flows for complex geometries.
Subgrid-Scale
Diffusivity: Wall Behavior and Dynamic Methods
Authors: Brillant Guillaume , Husson Sabine , Bataille Françoise
Published: May 2006

This study concerns the near-wall behavior of the subgrid-scale diffusivity. This is shown to depend on the thermal boundary conditions. Therefore, the constant subgrid-scale Prandtl number hypothesis is questionable and a direct modeling of the subgrid-scale diffusivity is considered instead. Large-eddy simulations are carried out using the Trio U code in a turbulent channel flow configuration with the three classical thermal boundary conditions (constant temperature, constant heat flux, and adiabatic wall). Different dynamic methods are used to model the subgrid-scale diffusivity and results are compared with constant subgrid-scale Prandtl number large-eddy simulations and with direct numerical simulations.
Linear Stability Analysis of the Effects of Camber and Blade Thickness on Cavitation Instabilities in Inducers
Authors: Horiguchi Hironori , Semenov Yury , Nakano Masataka , Tsujimoto Yoshinobu
Published: May 2006

It has been shown by experimental and numerical studies that various cavitation instabilities occur in inducers for rocket engines when the cavity length exceeds about 65% of the blade spacing. On the other hand, it has been pointed out by an experimental study that the cavitation instabilities occur when the pressure gradient near the throat becomes small to some degree. The present study is motivated to examine the latter criterion based on pressure gradient for cavitation instabilities from the viewpoint of theoretical analysis. For this purpose, analyses of steady flow and its stability were carried out for cavitating flow in cascades with circular arc and plano-convex blades by a singularity method based on closed cavity model. It was found that the criterion based on the cavity length for the occurrence of cavitation instabilities is more adequate than the criterion based on the pressure gradient. It was also found that the steady cavity length and the stability of the flow in both cascades can be practically correlated with a parameter σ∕[2(α−α0)], where σ is a cavitation number, α is an angle of attack, and α0 is a shockless angle of attack.
Experimental Investigation of Unstably Stratified Buoyant Wakes
Authors: Kraft Wayne N. , Andrews Malcolm J.
Published: May 2006

A water channel has been used as a statistically steady experiment to investigate the development of a buoyant plane wake. Parallel streams of hot and cold water are initially separated by a splitter plate and are oriented to create an unstable stratification. At the end of the splitter plate, the two streams are allowed to mix and a buoyancy-driven mixing layer develops. The continuous, unstable stratification inside the developing mixing layer provides the necessary environment to study the buoyant wake. Downstream a cylinder was placed at the center of the mixing layer. As a result the dynamic flows of the plane wake and buoyancy-driven mixing layer interact. Particle image velocimetry and a high-resolution thermocouple system have been used to measure the response of the plane wake to buoyancy driven turbulence. Velocity and density measurements are used as a basis from which we describe the transition, and return to equilibrium, of the buoyancy-driven mixing layer. Visual observation of the wake does not show the usual vortex street associated with a cylinder wake, but the effect of the wake is apparent in the measured vertical velocity fluctuations. An expected peak in velocity fluctuations in the wake is found, however the decay of vertical velocity fluctuations occurs at a reduced rate due to vertical momentum transport into the wake region from buoyancy-driven turbulence. Therefore for wakes where buoyancy is driving the motion, a remarkably fast recovery of a buoyancy-driven Rayleigh-Taylor mixing in the wake region is found.
Influence of Surface Roughness on the Aerodynamic Losses of a Turbine Vane
Authors: Zhang Qiang , Goodro Matt , Ligrani Phillip M. , Trindade Ricardo , Sreekanth Sri
Published: May 2006

The effects of surface roughness on the aerodynamic performance of a turbine vane are investigated for three Mach number distributions, one of which results in transonic flow. Four turbine vanes, each with the same shape and exterior dimensions, are employed with different rough surfaces. The nonuniform, irregular, three-dimensional roughness on the tested vanes is employed to match the roughness which exists on operating turbine vanes subject to extended operating times with significant particulate deposition on the surfaces. Wake profiles are measured for two different positions downstream the vane trailing edge. The contributions of varying surface roughness to aerodynamic losses, Mach number profiles, normalized kinetic energy profiles, Integrated Aerodynamics Losses (IAL), area-averaged loss coefficients, and mass-averaged loss coefficients are quantified. Total pressure losses, Mach number deficits, and deficits of kinetic energy all increase at each profile location within the wake as the size of equivalent sandgrain roughness increases, provided the roughness on the surfaces is uniform. Corresponding Integrated Aerodynamic Loss IAL magnitudes increase either as Mach numbers along the airfoil are higher, or as the size of surface roughness increases. Data are also provided which illustrate the larger loss magnitudes which are present with flow turning and cambered airfoils, than with symmetric airfoils. Also described are wake broadening, profile asymmetry, and effects of increased turbulent diffusion, variable surface roughness, and streamwise development.
Airfoil Performance at Low Reynolds Numbers in the Presence of Periodic Disturbances
Authors: Yarusevych S. , Kawall J. G. , Sullivan P. E.
Published: May 2006

The boundary-layer separation and wake structure of a NACA 0025 airfoil and the effect of external excitations in presence of structural vibrations on airfoil performance were studied experimentally. Wind tunnel experiments were carried out for three Reynolds numbers and three angles of attack, involving hot-wire measurements and complementary surface flow visualization. The results establish that external acoustic excitation at a particular frequency and appropriate amplitude suppresses or reduces the separation region and decreases the airfoil wake, i.e., produces an increase of the lift and∕or decrease of the drag. The acoustic excitation also alters characteristics of the vortical structures in the wake, decreasing the vortex length scale and coherency. Optimum excitation frequencies were found to correlate with the fundamental frequencies of the naturally amplified disturbances in the separated shear layer. The results suggest that acoustic waves play a dominant role in exciting the separated shear layer of the airfoil. Moreover, low-frequency structural vibrations are found to have a significant effect on airfoil performance, as they enhance the sound pressure levels within the test section.
Evaluation of GafChromic EBT prototype B for external beam dose verification
Authors: Todorovic M. , Fischer M. , Cremers F. , Thom E. , Schmidt R.
Published: May 2006

The capability of the new GafChromic EBT prototype B for external beam dose verification is investigated in this paper. First the general characteristics of this film (dose response, postirradiation coloration, influence of calibration field size) were derived using a flat‐bed scanner. In the dose range from , the sensitivity of the EBT prototype B film is ten times higher than the response of the GafChromic HS, which so far was the GafChromic film with the highest sensitivity. Compared with the Kodak EDR2 film, the response of the EBT is higher by a factor of 3 in the dose range from . The GafChromic EBT almost does not show a temporal growth of the optical density and there is no influence of the chosen calibration field size on the dose response curve obtained from this data. A MatLab program was written to evaluate the two‐dimensional dose distributions from treatment planning systems and GafChromic EBT film measurements. Verification of external beam therapy (SRT, IMRT) using the above‐mentioned approach resulted in very small differences between the planned and the applied dose. The GafChromic EBT prototype B together with the flat‐bed scanner and MatLab is a successful approach for making the advantages of the GafChromic films applicable for verification of external beam therapy.
Screen optics effects on detective quantum efficiency in digital radiography: Zero‐frequency effects
Authors: Lubinsky A. R. , Zhao Wei , Ristic Goran , Rowlands J. A.
Published: May 2006

Indirect flat panel imagers have been developed for digital radiography, fluoroscopy and mammography, and are now in clinical use. Screens made from columnar structured cesium iodide scintillators doped with thallium have been used extensively in these detectors. The purpose of this article is to investigate the effect of screen optics, e.g., light escape efficiency versus depth, on gain fluctuation noise, expressed as the Swank factor. Our goal is to obtain results useful in optimizing screens for digital radiography systems. Experimental measurements from structured samples were used to derive their screen optics properties, and the same methods can also be applied to powder screens. screens, all of the same thickness but with different optical designs and manufacturing techniques, were obtained from Hamamatsu Photonics Corporation. The pulse height spectra (PHS) of the screens were measured at different x‐ray energies. A theoretical model was developed for the light escape efficiency and a method for deriving light escape efficiency versus depth from experimental PHS measurements was implemented and applied to the screens. The results showed that the light escape efficiency varies essentially linearly as a function of depth in the samples, and that the magnitude of variation is relatively small, leading to a high Swank factor.
Optimization of a tomosynthesis system for the detection of lung nodules
Authors: Pineda Angel R. , Yoon Sungwon , Paik David S. , Fahrig Rebecca
Published: May 2006

Mathematical observers that track human performance can be used to reduce the number of human observer studies needed to optimize imaging systems. The performance of human observers for the detection of a lung nodule in anatomical backgrounds was measured as a function of varying tomosynthetic angle and compared with mathematical observers. The human observer results showed a dramatic increase in the percent of correct responses, from 80% in the projection images to 96% in the projection images with a tomosynthetic angle of just 3 degrees. This result suggests the potential usefulness of the scanned beam digital x‐ray system for this application. Given the small number of images (40) used per tomosynthetic angle and the highly nonstationary statistical nature of the backgrounds, the nonprewhitening eye observer achieved a higher performance than the channelized Hotelling observer using a Laguerre‐Gauss basis. The channelized Hotelling observer with internal noise and the eye filter matched to the projection data were shown to track human performance as the tomosynthetic angle changed. The validation of these mathematical observers extends their applicability to the optimization of tomosynthesis systems.
An abdominal aortic aneurysm segmentation method: Level set with region and statistical information
Authors: Zhuge Feng , Rubin Geoffrey D. , Sun Shaohua , Napel Sandy
Published: May 2006

We present a system for segmenting the human aortic aneurysm in CT angiograms (CTA), which, in turn, allows measurements of volume and morphological aspects useful for treatment planning. The system estimates a rough “initial surface,” and then refines it using a level set segmentation scheme augmented with two external analyzers: The global region analyzer, which incorporates aprioriknowledge of the intensity, volume, and shape of the aorta and other structures, and the local feature analyzer, which uses voxel location, intensity, and texture features to train and drive a support vector machine classifier. Each analyzer outputs a value that corresponds to the likelihood that a given voxel is part of the aneurysm, which is used during level set iteration to control the evolution of the surface. We tested our system using a database of 20 CTA scans of patients with aortic aneurysms. The mean and worst case values of volume overlap, volume error, mean distance error, and maximum distance error relative to human tracing were(s.d.);,(s.d.);,(s.d.);, and(s.d.);, respectively. When implemented on aPentium IV personal computer, the mean time required for segmentation was(s.d.). We also performed experiments that suggest that our method is insensitive to parameter changes within 10% of their experimentally determined values. This preliminary study proves feasibility for an accurate, precise, and robust system for segmentation of the abdominal aneurysm from CTA data, and may be of benefit to patients with aortic aneurysms.
Lower Cambrian Vendobionts from China and Early Diploblast Evolution
Authors: Shu D.-G. , Morris S. Conway , Han J. , Li Y. , Zhang X.-L. , Hua H. , Zhang Z.-F. , Liu J.-N. , Guo J.-F. , Yao Y. , Yasui K.
Published: May 2006

Ediacaran assemblages immediately predate the Cambrian explosion of metazoans and should have played a crucial role in this radiation. Their wider relationships, however, have remained refractory and difficult to integrate with early metazoan phylogeny. Here, we describe a frondlike fossil,
Stromatoveris
(
S. psygmoglena
sp. nov.), from the Lower Cambrian Chengjiang Lagerstätte (Yunnan, China) that is strikingly similar to Ediacaran vendobionts. The exquisite preservation reveals closely spaced branches, probably ciliated, that appear to represent precursors of the diagnostic comb rows of ctenophores. Therefore, this finding has important implications for the early evolution of this phylum and related diploblasts, some of which independently evolved a frondose habit.