Vaginal natural orifice transvaginal endoscopic surgery (vNOTES) surgical staging for endometrial carcinoma: The feasibility of an innovative approach.

OBJECTIVE: This study aims to describe the feasibility of treating early-stage endometrial cancer with hysterectomy, bilateral salpingo-oophorectomy, sentinel lymph node biopsy, and bilateral pelvic lymphadenectomy by vaginal Natural Orifice Transluminal Endoscopic Surgery (vNOTES). MATERIALS AND METHODS: A longitudinal study of prospectively registered patients was conducted at an academic tertiary care center. 15 patients who underwent vNOTES surgical staging of early endometrial carcinoma between January 2014 and December 2020 were included in the study. RESULTS: 15 patients between 20 and 80yrs of age with histologically proven Stage1 Gr1-2 endometrial cancer underwent vNOTES surgical staging. The mean age of the study population was 52.8 years (Standard Deviation [SD] 6.8) and the mean BMI was 27.8 kg/m2 (SD 6.4). The average operative time was 231.4 min (SD 41.0) with the mean estimated blood loss of 122.0 mL (SD 104.4). A total of 12 (80%) patients underwent SNL biopsy with ICG guided system, whereas 3 (20%) had pelvic lymph node dissection. There was one case with the surgical complication of bladder injury requiring conversion to conventional laparoscopy. CONCLUSION: With this study, we studied the feasibility of vNOTES surgery for early-stage endometrial cancer with minimal complications and the best long-term surgical outcome. The surgeries were performed by a single skilled endoscopist surgeon with previous experience with vNOTES surgery for adnexal tumors and hysterectomy. Our results showed the practicality of vNOTES in staging surgery for early-stage endometrial cancer. However, application to a larger cohort is required for more extensive surgical outcome studies.

The effect of the electronic platform of menopausal health screen system and counseling intervention on the empowerment of menopausal women: a quasi-experimental study.

This article explores the efficacy of the electronic platform of menopausal health screen system (EPMHSS) and counseling intervention on the empowerment of menopausal women, seventy four participants were randomly assigned to this study. The intervention group significantly relieved menopausal disturbance, reduced uncertainty, increased health behaviors, and decreased waist circumference after the fourth and eighth week compared with the control group. Our results proved that EPMHSS and counseling would help menopausal women to become more aware of their health, as a result, effectively empowered themselves to take actions for improving their health.

Vallecular cyst: a dangerous cause of failure to thrive in infants.

A 2-month-old full-term female infant with medical history of situs inversus totalis presented to the emergency department with congestion and abnormal breathing. She was discovered to have failure to thrive (FTT) and subsequently admitted. Investigations revealed a large vallecular mass at the base of her tongue which was noted to cause severe, intermittent airway obstruction. The mass underwent marsupialisation by otolaryngology (ENT) and pathology confirmed a diagnosis of vallecular cyst. The patient made a full recovery and is now growing and thriving. This case emphasises the need to consider anatomic airway abnormalities in the differential diagnosis of young infants with the constellation of respiratory symptoms and FTT. Such airway abnormalities can cause life-threatening airway obstruction if not discovered.

Novel Neovaginoplasty Using Rudimentary Uterine Horn Serosa and Pelvic Peritoneum as a Graft in Müllerian Anomalies with Vaginal Agenesis.

STUDY OBJECTIVE: To study the outcome of a novel method of laparoscopic neovaginal reconstruction using rudimentary uterine horn serosa and the pelvic peritoneum as a graft. DESIGN: Canadian Task Force classification II-1. SETTING: A university hospital. PATIENTS: A retrospective study of 14 patients from 2000 to 2014 of patients with vaginal agenesis who underwent laparoscopic neovagina reconstruction using rudimentary uterine horn serosa and the pelvic peritoneum as a graft. INTERVENTION: Patients with vaginal agenesis associated with müllerian agenesis who requested surgery. Tertiary referral center and laparoscopic unit. The creation of a neovagina using rudimentary uterine horn serosa and the pelvic peritoneum as a graft via a combined laparoscopic and vaginal route. MEASUREMENTS AND MAIN RESULTS: Data were collected retrospectively including postoperative vaginal length and width, complications, stenosis or reoperations, dyspareunia, and sexual satisfaction. There were no major complications from the surgery with no rectal perforation or bladder or ureteric injury. The postoperative mean (±SD) vaginal length was 6.0±0.7 cm and a width of 2 fingerbreadths. The mean operation time was 142.7±45.9 min. Median blood loss was 100 ml (range: 10 to 300 mL). The mean duration of the hospital stay was 6.6±1.6 days. The follow-up period ranged from 3 to 84 months with a median follow-up of 11 months. CONCLUSION: Lee's method of neovaginoplasty using rudimentary uterine horn serosa and the pelvic peritoneum as a graft is a good method for neovagina creation with minimal morbidity, fast recovery, and minimal complications. This method results in good anatomic and functional outcome and can be a method that is widely used.

Relationship between the social support and self-efficacy for function ability in patients undergoing primary hip replacement.

BACKGROUND: The World Health Organization (WHO) reported that nearly 25% of people will suffer from physical disability owing to the bone and joint problems until 2050. The condition of patients with this type of difficulty could be improved by increasing positive self-efficacy and instigating suitable medical treatment to implement self-efficacy for functional ability (SEFA) and physical functional ability self-care. In this study, we aim to evaluate the influence of social support on SEFA in patients after total hip arthroplasty. METHODS: This cross-sectional study used structural questionnaires, telephone appointments, and data collection to obtain patient characteristics, such as gender, age, educational level, and marital status. Questionnaires about social support and self-efficacy for functional ability (SEFA) were sent to 200 patients at 3 months following a primary total hip replacement from September 2011 to December 2014. Factor analysis was used to categorize the dimensions of social support; the t test, analysis of variance (ANOVA), and correlation analysis were applied to screen factors influencing SEFA. Multiple regression analysis was employed to ascertain the relationships between patient characteristics, social support, and SEFA. RESULTS: In total, 134 patients responded to the questionnaires. Lower SEFA scores were observed in patients of an older age, unmarried patients, and those with a low level of education. Correlation analysis showed that emotional information and appraisal support, instrumental support, and SEFA were positively correlated. Multiple regression analysis was applied to ascertain the relationships between patient characteristics, social support, and SEFA. We identified significant coefficient values of - 0.187 for age, 5.344 for emotional information and appraisal support, and 1.653 for instrumental support. CONCLUSION: The results of this study demonstrated that in patients undergoing primary hip replacement, positive impacts on SEFA were observed in relation to emotional information, appraisal support and instrumental support. The results indicated that enhancing emotional information and appraisal support could improve a patient's self-efficacy for functional ability.

Surgical and survival outcomes of laparoscopic staging surgery for patients with stage I ovarian cancer.

OBJECTIVE: To assess the feasibility and survival outcomes of laparoscopic staging for patients with stage I ovarian cancer. MATERIALS AND METHODS: Consecutive patients who underwent laparoscopic staging surgery for stage I ovarian cancer from January 2002 to December 2014 were evaluated retrospectively by chart review. RESULTS: Twenty-four patients with mean age 43.9 ± 9.9 years and mean body mass index 24.0 ± 3.8 kg/m2 were included, in which 12 (50%) patients were in stage IA and 12 (50%) in stage IC. The histological types included serous in 6 (25%), mucinous in 7 (29.1%), endometrioid in 6 (25%), clear cell in 5 (20.8%) patients. The mean surgical time was 306.4 ± 98.5 min, and the mean blood loss was 204.2 ± 188.6 mL. None of the patients required conversion to laparotomy. The median numbers of resected pelvic and para-aortic lymph nodes were 20 and 4, respectively. One (4.1%) patient encountered bowel injury intraoperatively, and the other 1 (4.1%) patient hydronephrosis postoperatively. The overall survival rate was 95% in the current series in a median follow-up of 31.5 months. CONCLUSION: Laparoscopic staging surgery performed for early stage ovarian cancer has better long term survival outcomes than the literature report. Laparoscopic treatment by a trained gynecologic oncologist is an ideal alternative for early stage ovarian cancer with the advantage of minimal invasiveness.

Severe infections in patients with lupus nephritis treated with immunosuppressants: A retrospective cohort study.

AIM: Lupus nephritis (LN) is associated with significant morbidity and mortality and hence usually treated aggressively with immunosuppressants. This predisposes LN patients to increased infections, yet few studies have evaluated LN patients for infective complications. We aimed to describe the epidemiology and identify risk factors for infections requiring hospitalization among patients with biopsy-proven LN. METHODS: This was a single-centre retrospective cohort study of patients with biopsy-proven LN between 1 January 2000 and 31 May 2009. Patients were excluded if they were <16 years old at time of biopsy, had previous kidney transplant or if pharmacotherapy data were incomplete. Hospitalizations for infections, bacteraemia and polymicrobial infections were recorded until patients' last visit or when they received immunosuppression for non-glomerulonephritis indications, such as solid organ transplant or chemotherapy. RESULTS: We studied 189 patients who had biopsy-proven lupus nephritis. Median age at diagnosis was 36.9 (IQR: 27.4, 47.5) years and 82% were female. Most patients received at least one immunosuppressant after LN diagnosis, including glucocorticosteroids in 94.2%. One hundred and four patients (60.3%) had at least one hospitalization for infection at 11 (1, 53) months from diagnosis. Bacteraemia occurred in 26 patients (13.8%) and 32 patients (16.9%) had polymicrobial infections. On multivariate analysis, LN relapse was associated with hospitalization for infection (OR 2.33 (1.18, 4.60), P = 0.01) and bacteraemia (OR 3.47 (1.05, 11.45), P = 0.04). Infection-related mortality occurred in 10 patients (5.3%). CONCLUSION: Serious infections are common among patients with LN and are associated with mortality.

Differing Requirements for MALT1 Function in Peripheral B Cell Survival and Differentiation.

During a T cell-dependent immune response, formation of the germinal center (GC) is essential for the generation of high-affinity plasma cells and memory B cells. The canonical NF-κB pathway has been implicated in the initiation of GC reaction, and defects in this pathway have been linked to immune deficiencies. The paracaspase MALT1 plays an important role in regulating NF-κB activation upon triggering of Ag receptors. Although previous studies have reported that MALT1 deficiency abrogates the GC response, the relative contribution of B cells and T cells to the defective phenotype remains unclear. We used chimeric mouse models to demonstrate that MALT1 function is required in B cells for GC formation. This role is restricted to BCR signaling where MALT1 is critical for B cell proliferation and survival. Moreover, the proapoptotic signal transmitted in the absence of MALT1 is dominant to the prosurvival effects of T cell-derived stimuli. In addition to GC B cell differentiation, MALT1 is required for plasma cell differentiation, but not mitogenic responses. Lastly, we show that ectopic expression of Bcl-2 can partially rescue the GC phenotype in MALT1-deficient animals by prolonging the lifespan of BCR-activated B cells, but plasma cell differentiation and Ab production remain defective. Thus, our data uncover previously unappreciated aspects of MALT1 function in B cells and highlight its importance in humoral immunity.

pH-controlled multiple chiral inversion that induces molecular dimerization in a gold(I)-cobalt(III) coordination system with L-cysteinate.

Herein, a unique coordination system that exhibits multiple chiral inversions and molecular dimerization in response to a subtle pH change is reported. Treatment of (Δ)2-H3[Au3Co2(L-cys)6] (H3[1 a]) with [Co3(aet)6](NO3)3 (aet=2-aminoethanethiolate) in water at pH 7 gave a 1:1 complex salt of [Co3(aet)6](3+) and [1 a](3-), retaining the Au(I)3Co(III)2 structure and chiral configurations of [1 a](3-). Similar treatment at pH 9 led to not only the inversion of all of the chiral Co(III) and S centers but also the dimerization of [1 a](3-), giving a 2:1 complex salt of [Co3(aet)6](3+) and (Λ)4(R)12-[Au6Co4(L-cys)12](6-) ([2](6-)). When dissociated from [Co3(aet)6](3+) in solution, [2](6-) was converted to (Λ)2(R)6-[Au3Co2(L-cys)6](3-) ([1 b](3-)) with retention of the chiral configurations.

Long-term survival outcome of laparoscopic staging surgery for endometrial cancer in Taiwanese experience.

OBJECTIVE: This study aims to evaluate the long-term safety and efficacy of laparoscopic staging surgery (LSS) for endometrial cancer in Taiwanese women. MATERIALS AND METHODS: This is a longitudinal study of prospectively registered 105 patients who underwent LSS for endometrial cancer between June 1995 and June 2008. RESULTS: The mean duration of surgery was 186.8 minutes, and the mean intraoperative blood loss was 220.38 mL. The median number of retrieved pelvic lymph nodes was 18. The intraoperative complication rate was 4.8%, including two cases of ureteral injury and one case each of bladder injury, bowel injury, and vascular injury. No patient required conversion to laparotomy. During the median follow-up of 55.3 months, six cases of recurrence (5.7%) and three tumor-related deaths (2.9%) were recorded. The 5-year disease-free survival and the overall survival were 93.39% and 98.05%, respectively. CONCLUSION: The study revealed favorable perioperative outcomes and better long-term survival than reported in the Taiwan Cancer Registry, and similar good surgical results to those reported in the Western studies. Therefore, LSS by experienced surgeons for endometrial cancer is a feasible and efficacious alternative to laparotomy.

Chromosome X-encoded Cancer/Testis antigens are less frequently expressed in non-seminomatous germ cell tumors than in seminomas.

Cancer/Testis (CT) antigens are normally only expressed in germ cells and yet are aberrantly activated in a wide variety of human cancers. Most chromosome X-encoded CT antigens (CT-X) show restricted expression in pre-meiotic germ cells in adult testis, except for the expression of SPANX in post-meiotic germ cells. In the present study, the expression of eight CT-X antigens (MAGE-A, NY-ESO-1, GAGE, MAGE-C1/CT7, MAGE-C2/CT10, CT45, SAGE1, and SPANX) in non-seminomatous germ cell tumors was evaluated immunohistochemically, including 24 embryonal carcinomas, 20 yolk sac tumors, 9 teratomas, and 3 choriocarcinomas, and the results were compared to our previous study of 77 classic seminomas and 2 spermatocytic seminomas. SPANX was not detected in any germ cell tumors tested. Spermatocytic seminoma showed strong expression of all CT-X antigens tested (except SPANX), reflecting their origin from adult CT-Xpositive pre-meiotic germ cells. Classic seminomas, originating from prenatal gonocytes, showed widely variable frequency of CT-X antigen expression, ranging from > 80% (CT7, CT10, CT45, and GAGE), 63% (MAGE-A), 18% (NY-ESO-1) to only 4% (SAGE1). In comparison, non-seminomatous germ cell tumors expressed CT-X antigens much less frequently and usually only in small subsets of tumor cells. Intratubular germ cell neoplasia (ITGCN) were mostly CT-X-negative, even in CT-X positive classic seminomas. These findings indicate that CT-X antigens are not expressed in the fetal precursor cells for germ cell tumors, and their expression likely reflects germ cell differentiation of the neoplastic cells (in seminomas) or aberrant gene activation as cancer antigens (in non-seminomatous tumors).

Effects of CT-Xp gene knock down in melanoma cell lines.

Cancer/testis (CT) genes are encoded by genes that are normally expressed only in the human germ line but which are activated in various malignancies. CT proteins are frequently immunogenic in cancer patients and their expression is highly restricted to tumors. They are thus important targets for anticancer immunotherapy. In several different tumor types, the expression of CT-X genes is associated with advanced disease and poor outcome, indicating that their expression might contribute to tumorigenesis. CT-X genes encoding members of the MAGE protein family on Xq28 have been shown to potentially influence the tumorigenic phenotype. We used small interfering RNA (siRNA) to investigate whether CT-X mapping to the short arm of the X-chromosome might also have tumorigenic properties and therefore be potentially targeted by functional inhibitors in a therapeutic setting. siRNAs specific to GAGE, SSX and XAGE1 were used in cell proliferation, migration and cell survival assays using cell lines derived from melanoma, a tumor type known to present high frequencies of expression of CT antigens. We found that of these, those specific to GAGE and XAGE1 most significantly impeded melanoma cell migration and invasion and those specific to SSX4 and XAGE1 decreased the clonogenic survival of melanoma cells. Our results suggest that GAGE, XAGE1 and SSX4 might each have a role in tumor progression and are possible therapeutic targets for the treatment of melanoma and other malignancies.

Mediating effects of aryl-hydrocarbon receptor and RhoA in altering brain vascular integrity: the therapeutic potential of statins.

We have demonstrated previously that focal adhesion kinase (FAK)/RhoA alteration by the aryl-hydrocarbon receptor (AhR) agonist 3-methylcholanthrene (3MC) is involved in the antimigratory effects of 3MC in human umbilical vascular endothelial cells. Here, we identified that signaling properties and molecular mechanisms of RhoA/ß-catenin were both implicated in alterations to blood-brain barrier integrity. The mechanisms of action were the down-regulation of integrin, the extracellular matrix, and adherens junction stability. PTEN phosphorylation by 3MC-mediated AhR/RhoA activation increased the proteasomal degradation of ß-catenin through PKCδ/pGSK3ß-mediated ß-catenin phosphorylation; the crucial roles of AhR/RhoA in this process were verified by using gain- or loss-of-function experiments. The decrease in ß-catenin led to decreased expression of fibronectin and α5ß1 integrin. Additionally, protein interactions among FAK, VE-cadherin, vinculin, and ß-actin were simultaneously decreased, resulting in adherens junction instability. Novel functional TCF/LEF1 binding sites in the promoter regions of fibronectin and α5/ß1 integrin were identified by electrophoretic mobility shift and chromatin immunoprecipitation assays. The results indicate that the binding activities of ß-catenin decreased in mouse cerebrovascular endothelial cells treated with 3MC. In addition, simvastatin and pravastatin treatment reversed 3MC-mediated alterations in mouse cerebrovascular endothelial cells by RhoA inactivation, and the in vitro findings were substantiated by an in vivo blood-brain barrier assay. Thus, endothelial barrier dysfunction due to 3MC occurs through AhR/RhoA-mediated ß-catenin down-regulation, which is reversed by simvastatin treatment in vivo.

Chromosome X-encoded cancer/testis antigens show distinctive expression patterns in developing gonads and in testicular seminoma.

BACKGROUND: Cancer/testis (CT) antigens are cancer antigens normally expressed in adult testicular germ cells. The expression of chromosome X-encoded CT antigens (CT-X antigens) in human fetal gonads and in testicular seminomas was examined. METHODS: The expression of 10 CT-X antigens (MAGEA, NY-ESO-1, GAGE, CT7/MAGEC1, CT10/MAGEC2, CT45, SAGE1, SSX2, NXF2 and SPANX) was studied immunohistochemically. RESULTS: In adult human testis, SPANX is expressed in late spermatids and spermatozoa, whereas all other CT-X antigens are predominantly expressed in spermatogonia or primary spermatocytes. All CT-X antigens except SPANX are expressed in human fetal germ cells. CT-X-positive germ cells appear as early as 13 weeks after gestation, increase with age and reach a plateau at around 22 weeks. In the fetal ovary, CT-X-positive oogonia are most abundant at around 24 weeks and sharply decrease subsequently. CT-X antigens are almost exclusively expressed in OCT3/4-negative gonocytes and their expression appears to coincide with the loss of pluripotency. Spermatocytic seminoma, a neoplasm derived from adult pre-meiotic germ cells, showed uniform expression of all CT-X antigens except SPANX. In contrast, most seminomas (>80%) express CT7, CT45, GAGE and CT10 but express MAGEA, NXF2 and NY-ESO-1 at lower frequency, and very rarely express SSX2 and SAGE1. CONCLUSIONS: Most CT-X antigens are expressed in human fetal germ cells after they have lost stem cell characteristics, with predominant expression in pre-meiotic germ cells. Spermatocytic seminomas showed expression of all CT-X antigens except SPANX, whereas classical seminomas only express some CT-X antigens, reflecting their different origins from adult versus fetal germ cells.

Multiple cancer/testis antigens are preferentially expressed in hormone-receptor negative and high-grade breast cancers.

BACKGROUND: Cancer/testis (CT) antigens are protein antigens normally expressed only in germ cells of testis, and yet are expressed in a proportion of a wide variety of human cancers. CT antigens can elicit spontaneous immune responses in cancer patients with CT-positive cancers, and CT antigen-based therapeutic cancer vaccine trials are ongoing for "CT-rich" tumors. Although some previous studies found breast cancer to be "CT-poor", our recent analysis identified increased CT mRNA transcripts in the ER-negative subset of breast cancer. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we performed a comprehensive immunohistochemical study to investigate the protein expression of eight CT genes in 454 invasive ductal carcinomas, including 225 ER/PR/HER2-negative (triple-negative) carcinomas. We found significantly more frequent expression of all eight CT antigens in ER-negative cancers, and five of them--MAGEA, CT7, NY-ESO-1, CT10 and CT45, were expressed in 12-24% of ER-negative cancers, versus 2-6% of ER-positive cancers (p<0.001 to 0.003). In comparison, GAGE, SAGE1 and NXF2 were only expressed in 3-5% of ER-negative and 0-2% of ER-positive cancers. ER-negative cancers were also more likely to simultaneously co-express multiple CT antigens, with 27% (34/125) of ER-negative, CT-positive tumors expressing three or more CT antigens. HER2 status had no consistent effect on CT expression, and triple-negative carcinomas showed similar frequencies of MAGEA and NY-ESO-1 expression as ER-negative/HER2-positive carcinomas. More frequent CT expression was also found in tumors with higher nuclear grade (p<0.001 to p = 0.01) and larger in size (>2 cm). CONCLUSIONS/SIGNIFICANCE: CT antigens are preferentially expressed in hormone receptor-negative and high-grade breast cancer. Considering the limited treatment options for ER/PR/HER2 triple-negative breast cancer, the potential of CT-based immunotherapy should be explored.

Ectopic overexpression of haem oxygenase-1 protects kidneys from carboplatin-mediated apoptosis.

BACKGROUND AND PURPOSE: We previously reported that the activation of the nuclear factor of activated T-lymphocyte-3 (NFAT3) by carboplatin leads to renal apoptosis as a result of oxidative stress, which is reversed by N-acetylcysteine. Herein, we extend our previous work to provide evidence of the molecular mechanisms of haem oxygenase (HO)-1 in protecting against injury. EXPERIMENTAL APPROACH: Protective mechanisms of HO-1 in carboplatin-mediated renal apoptosis were examined in C57BL/6 mice and rat renal tubular cells (RTC) with HO-1 induction or inactivation/knockdown. KEY RESULTS: The HO-1, induced by cobalt protoporphyrin, protected against carboplatin-induced renal injury in vivo. This protection was decreased by an inhibitor of HO-1 action, tin protoporphyrin. In cultures of RTC, carboplatin-induced apoptosis was similarly affected by HO-1 overexpression or knockdown. Carboplatin-mediated NFAT3 activation and apoptosis involve activation of the signalling kinases, extracellular signal regulated kinase, Jun N-terminal kinase and protein kinase C, and such activation was reversed in cells overexpressing HO-1. Both products of the HO-1 reaction, CO and bilirubin, inhibited (by 30-40%) NFAT3 activation and production of the pro-apoptotic proteins Bcl-XS/Bax. Additionally, the activation of NFκB was markedly decreased by HO-1 induction. CONCLUSION AND IMPLICATIONS: HO-1 and its reaction products show anti-apoptotic effects in carboplatin-mediated renal injury. A novel functional NFAT3 binding site identified in the rat HO-1 promoter region was involved in producing a 1.5-fold to 2.5-fold increase in HO-1 induction by carboplatin. Nevertheless, only HO-1 overexpression and activation prior to the carboplatin challenge provided protection against carboplatin-induced injury.

Long-term survival outcomes of laparoscopically assisted radical hysterectomy in treating early-stage cervical cancer.

OBJECTIVE: The objective of the study was to determine the long-term disease-free and overall survival outcomes of laparoscopic treatment of early-stage cervical cancer. STUDY DESIGN: This was a longitudinal study of prospectively registered patients of cervical cancer undergoing laparoscopic surgery from June 1994 to December 2005. RESULTS: A total of 139 patients were included, in which 60 patients were in International Federation of Gynecology and Obstetrics stage IA, 76 in IB, and 3 in IIA. Mean operation time was 231.1 +/- 6.1 minutes. Median number of pelvic lymph node retrieval was 16. Major intraoperative complications included 1 great vessel injury, 1 ureteral injury, 1 colon injury, and 6 cystotomies. In a median follow-up of 92.1 months, the mean +/- SEM cumulative disease-free and overall survival rates were 91.01% +/- 2.77% and 92.78% +/- 3.06%, respectively. CONCLUSION: The laparoscopic approach has favorable long-term survival outcomes and perioperative morbidity. With the advantage of minimal invasiveness, laparoscopic treatment by experienced surgeons is an ideal alternative for early-stage cervical cancer.

Expression of cancer testis antigen CT45 in classical Hodgkin lymphoma and other B-cell lymphomas.

We have shown previously that cancer/testis (CT) antigen, CT45, is expressed in various epithelial cancers at a frequency of <5% to approximately 35%. In this study, the protein expression of CT45 was examined in non-Hodgkin B-cell lymphomas and classical Hodgkin lymphoma by immunohistochemical analysis. Serological response to CT45 was also evaluated by ELISA using CT45 recombinant protein and sera from patients with Hodgkin lymphoma. None of the 80 low-grade B-cell lymphomas, including chronic lymphocytic leukemia/small lymphocytic lymphoma, follicular lymphoma, and mantle cell lymphoma, expressed CT45. In comparison, CT45 was expressed in 28 of 126 (22%) diffuse large B-cell lymphomas (DLBCL). A remarkably high percentage (42/72, 58%) of classical Hodgkin lymphoma contained CT45-positive Reed-Sternberg cells. Nodular sclerosis and mixed-cellularity subtypes had similar frequency of CT45 expression, but most EBV-positive cases were CT45 negative. Gray-zone lymphoma (cases with features of both DLBCL and classical Hodgkin lymphoma) also showed frequent (64%) CT45 expression. Evaluation of reactive lymphoid tissues showed scattered CT45-positive lymphocytes in a single case of florid follicular hyperplasia, raising the possibility that this case was an evolving malignancy. Despite frequent CT45 expression, only 1 of 67 Hodgkin lymphoma patients had detectable anti-CT45 antibodies in the serum, suggesting that the immune response to CT45 may be suppressed. In conclusion, classical Hodgkin lymphoma has the highest frequency of CT45 expression among all malignancies tested to date, the frequency of CT45 expression in DLBCL is similar to that seen in epithelial cancers, and low-grade non-Hodgkin B-cell lymphomas do not express CT45.

Cancer/testis antigen CT45: analysis of mRNA and protein expression in human cancer.

CT45 is a cancer/testis gene that we previously identified by massively parallel signature sequencing. Encoded by a multigene family on chromosome X, CT45 showed restricted mRNA expression to normal testis and various cancers. In this study, monoclonal antibodies were generated against recombinant CT45 protein, and CT45 protein expression in normal and tumor tissues was evaluated by immunohistochemical analysis. In adult normal tissue, CT45 expression was restricted to testicular germ cells, detected as a nuclear protein mainly at the stage of primary spermatocytes. In tumors, CT45 protein expression correlated with the mRNA levels detected by quantitative RT-PCR, and most lung cancer and ovarian cancers with CT45 mRNA at levels >1% of testicular expression were CT45 protein-positive. In positive cases, CT45 showed expression patterns that ranged from diffuse strong staining to heterogeneous and patchy expression. In lung cancer, CT45 expression was least frequent in adenocarcinoma, more frequent in squamous cell carcinoma and neuroendocrine tumors. Using tissue microarrays, 376 lung cancer, 219 ovarian cancer and 155 breast cancer were evaluated for CT45 protein expression. The expression frequency was highest in ovarian cancer (37%), followed by lung cancer (13%) and lowest in breast cancer (<5%). Given the focal nature of CT45 expression in many cases, these numbers represented the minimal frequency of expression in these tumor types. In summary, the expression frequency and characteristics of CT45 expression are similar to other CT cancer vaccine targets currently in clinical trials, e.g., NY-ESO-1 and MAGE-A, suggesting CT45 as a potentially useful cancer target.

Expression of Globo H and SSEA3 in breast cancer stem cells and the involvement of fucosyl transferases 1 and 2 in Globo H synthesis.

We examined the expression in breast cancer stem cells (BCSCs) of Globo H, a potential tumor-associated antigen for immunotherapy of epithelial cancers including breast cancer. Flow cytometry revealed Globo H expression in 25/41 breast cancer specimens (61.0%). Non-BCSCs from 25/25 and BCSCs from 8/40 (20%) expressed Globo H. We showed the expression of stage-specific embryonic antigen 3 (SSEA3), the pentasaccharide precursor of Globo H, in 31/40 (77.5%) tumors. Non-BCSCs from 29/40 [corrected] and BCSCs from 25/40 (62.5%) expressed SSEA3. Like Globo H, SSEA3 expression in normal tissues was predominately at the secretory borders of epithelium, where access to the immune system is restricted. Immunization of mice with Globo H-KLH and alpha-GalCer induced antibodies reactive with Globo H and SSEA3, suggesting that a Globo H-based vaccine will target tumor cells expressing Globo H or SSEA3. We next sought to reduce Globo H expression by siRNA targeting fucosyltransferase (FUT) 1 and 2, which mediate alpha-1,2 linkage of fucose to SSEA3 to generate Globo H. We showed both genes to be involved in the biosynthesis of Globo H. Moreover, FUT2 expression in BCSCs was significantly lower than in non-BCSCs harvested from a primary human breast cancer in NOD/SCID mouse, whereas FUT1 was slightly lower in BCSCs. Thus, the lower expression of Globo H in BCSCs may be attributed to less FUT2/FUT1, and to reduced SSEA3 in BCSCs compared with non-BCSCs. Our findings provide insight into further development of a Globo H-based vaccine and FUT1/FUT2-targeted therapy for breast cancer.