Validation of the distress and impact thermometer and the changes of mood during the first 6 months of treatment in gynecological cancer patients: a Kansai Clinical Oncology Group (KCOG)-G1103 prospective study.

PURPOSE: To verify distress and impact thermometer (DIT) for screening emotional distress in gynecological cancer patients by Hospital Anxiety and Depression Scale total (HADS-T) as gold standard and to assess emotional changes by DIT and HADS-T. METHODS: A prospective study was conducted in newly diagnosed gynecological cancer patients during the peri-treatment period after the cancer diagnosis followed by 6-month. We defined a HADS-T score of ≥11 as being indicative of emotional distress. RESULTS: 117 patients were enrolled between May 1, 2011 and March 31, 2012, and 95 were eligible. The median age was 54 years (range 31-77). (1) From the baseline to 3-month, distress (DIT-D) ≥4 with Impact (DIT-I) ≥2 exhibited sensitivity, specificity, positive predictive value (PPV), and negative predictive values (NPV) of 0.776 [95 % confidential interval (CI) 0.688, 0.850], 0.889 (95 % CI 0.824, 0.954), 0.868 (95 % CI 0.792, 0.949), and 0.808 (95 % CI 0.731, 0.886), respectively. (2) At 6-month, DIT-D ≥2 with DIT-I ≥1 exhibited sensitivity, specificity, PPV and NPV of 0.893 (95 % CI 0.778, 1), 0.825 (95 % CI 0.707, 0.942), 0.781 (95 % CI 0.638, 0.928), and 0.917 (95 % CI 0.826, 1). (3) At 6-month, the HADS-T, DIT-D, and DIT-I scores in individual patients were significantly reduced by a mean of 4.57 (p < 0.0001), 2.34 (p < 0.0001), and 1.10 (p = 0.0031), respectively, compared with those scores of baseline (Student's paired t test), but still remained high. CONCLUSIONS: (1) On acute phase within 3-month setting, DIT; DIT-D ≥4 with DIT-I ≥2, is a reliable cut-off to screen emotional distress among gynecological cancer patients. (2) The patients' moods had improved, but not completely recovered at 6-month after the diagnosis.

Feasibility and efficacy of intraperitoneal docetaxel administration as salvage chemotherapy for malignant gynaecological ascites.

Ovarian and endometrial cancers diagnosed at advanced stages are often associated with malignant ascites. This study aimed to determine the safety, feasibility and efficacy of intraperitoneal (IP) docetaxel (TXT) for the treatment of ascites. A phase I study, including nine patients, was undertaken to determine the maximum tolerable dose. Efficacy was retrospectively assessed in 18 patients treated with 40-70 mg/m(2) IP TXT between 2005 and 2012. In a phase I study, the dose was safely escalated to a maximum of 70 mg/m(2), at which level no patients had grade -3 haematological adverse events. In a retrospective study of 18 patients, seven had an Eastern Cooperative Oncology Group performance status of 3; 16 had prior paclitaxel administration and two, with doses of 40 and 70 mg/m(2), experienced a serological response and a decrease in paracentesis. Thus, palliative treatment of recurrent OC should be further studied with 40 mg/m(2) among more patients, and 70 mg/m(2) could be evaluated for first-line IP chemotherapy.

Detection of unruptured ovarian pregnancy subsequently successfully treated by conservative laparoscopic surgery: a case report and review of the literature.

Early detection of ovarian pregnancy (OP) is essential for successful laparoscopic conservative surgery. However, early preoperative ultrasonography-based diagnosis is often difficult when fetal cardiac activity or the yolk sac is absent. The authors report a case of OP diagnosed at eight weeks gestational age in a natural pregnancy. The patient presented with amenorrhea and transient vaginal bleeding, and slight tenderness in the right ovary was noted during vaginal ultrasonography. Furthermore, ultrasonography showed a gestational sac (GS) without fetal cardiac activity or yolk sac, consistent with OP, and an adjacent compressible lutein cyst. The uterus, fallopian tubes, and left ovary were normal, and no cul-de-sac blood or ascites were found. Laparoscopy showed a two-cm mass partially covering the right ovary, which contained an unruptured GS. Subsequently, the mass was removed, and OP was histologically confirmed.

Possible presence of O-linked carbohydrate in the human male reproductive tract CD52.

Male reproductive tract CD52 (mrtCD52) is an antigen recognized by a complement-dependent sperm-immobilizing monoclonal antibody (SI-Abs) derived in an infertile patient. The molecule has been shown to contain a unique N-linked carbohydrate that does not cross-react with other tissues. In this study, we have investigated whether O-linked carbohydrate as well as N-linked carbohydrate is present in mrtCD52 using specific lectins and anti-CD52 core peptide antiserum. The lectin PNA, which recognizes O-linked carbohydrate [Galbeta1-3GalNAc], reacted with mrtCD52 and showed a similar polymorphic reaction pattern to that of the anti-peptide antiserum in western blotting analysis on two-dimensional SDS-PAGE. The PNA-reactive spots disappeared after removal of O-linked carbohydrate, but not after removal of N-linked carbohydrate. These results suggest that O-linked carbohydrate is present in mrtCD52. The moiety may possibly contribute to a specific antigenic epitope of mrtCD52.

Intravenous nedaplatin and intraarterial cisplatin with transcatheter arterial embolization for patients with locally advanced uterine cervical cancer.

Nedaplatin is a platinum analog that has less renal toxicity and higher efficacy for uterine cervical cancer than cisplatin. Intraarterial cisplatin has been shown to be more effective than intravenous cisplatin in the treatment of cervical cancer. To improve the prognosis of cervical cancer, we studied combination chemotherapy of intravenous nedaplatin and intraarticular cisplatin with transcatheter arterial embolization (TAE). The criteria for selecting patients for this study were as follows: age 16-75 years, stage Ib2-IV according to the classification of the International Federation of Gynecology and Obstetrics (FIGO), performance status between 0 and 2, a creatinine clearance of >40 ml/min, adequate bone marrow and adequate renal and hepatic function. Thirty-two patients, aged 29-72 years (median: 55) were treated. FIGO stage was Ib2 in seven patients, IIa in seven patients, IIb in four, IIIa in one, IIIb in seven and IVa in six. Twenty-four patients had squamous cell carcinoma, three had adenocarcinoma and five had adenosquamous carcinoma. Written informed consent was obtained from all patients. Nedaplatin (30-70 mg/m2) was administered intravenously on day 1 and cisplatin (70 mg/m2) was administered intraarticularly via both uterine arteries on day 3 using the Seldinger method. TAE was then performed. This course of treatment was repeated every 3 weeks for 2-3 cycles. Response to the therapy was defined by magnetic resonance imaging. Partial response was found in 59% patients (19/32) and complete response in 34% (11/32), with an overall response rate of 94% (30/32). Myelosuppression was manageable. No grade 2 neurotoxicity was observed. The median follow-up was 32 months (6-53 months), with 84% of patients showing an overall survival of 1 year and 77% showing an overall survival of 2 years. These results show that this combination chemotherapy effected a high response rate. However, its influence on long-term survival remains to be determined.

Multicentric reticulohistiocytosis in a patient with severe preeclampsia.

A multigravida patient with polyarthralgia and eruptions on the head and fingers was seen at 6 weeks' gestation. No histological examination was performed before the current pregnancy. She developed severe early onset preeclampsia associated with swelling of the knees and increased cutaneous nodules, biopsies of which revealed multicentric reticulohistiocytosis. At 28 weeks' gestation an elective cesarean section was performed and a 580-g male infant was delivered.

Expression of a recombinant porcine zona pellucida glycoprotein ZP1 in mammalian cells.

Porcine zona pellucida glycoprotein (pZP1) is a good candidate for a contraceptive vaccine. For the purpose of producing glycosylated pZP1, several types of recombinant pZP1 proteins were produced in mammalian cell lines. In the first experiment, a minigene encoding pZP1 (681 amino acids) was designed for insertion into an expression vector and then transfected to three cell lines (293T, CHO-K1, and LLC-PK1). The resulting recombinant proteins were highly glycosylated and were localized in the cytoplasm. To produce a secretory type of recombinant pZP1, in the second experiment, a cDNA coding for pZP1 excluding a putative transmembrane region and a smaller cDNA coding for 1-198 amino acid residues of pZP1 were designed to produce fusion proteins with the human IgG1 heavy chain. The resultant recombinant proteins were secreted into the supernatant from both transfected cell cultures. Recombinant secretory proteins are useful because of their simple affinity purification.

Blocking effect of antisera to recombinant zona pellucida proteins (r-ZPA) on in vitro fertilization.

PROBLEM: The zona pellucida is a good target antigen for contraceptive vaccines due to its strong immunogenicity and high tissue specificity. However, this contraceptive effect is inevitably associated with ovarian failure. Therefore, it is necessary to define an epitope of the zona antigen to which the antibody produced inhibits fertilization without any undesirable side effects. METHOD OF STUDY: The DNA fragment coding for the NH2-terminal region of porcine zona pellucida proteins (ZPA) (1-198 amino acids) and human ZPA (1-206 amino acids) was prepared to produce recombinant porcine ZPA (r-ZPA), r-pZPA1-198 and r-hZPA1-206. Using Freund's complete adjuvant. antisera against these proteins were raised in rabbits. RESULTS: The resultant antisera to r-pZPA1-198 and r-hZPA1-206 were cross-reacted with each other on enzyme-linked immunosorbent assay and immunofluorescent staining. The antiserum to r-pZPA1-198 inhibited in vitro fertilization in pigs, but not human sperm binding to the zona pellucida, while the antiserum to r-hZPA1-206 inhibited the human sperm-binding assay. CONCLUSIONS: Antiserum to r-ZPA inhibited fertilization in the animal species possessing a homologous amino-acid sequence as an immunogen. The recombinant protein, r-hZPA1-206 seems to be a feasible candidate for the development of contraceptive vaccines for humans.

Cloning of a cDNA coding for porcine zona pellucida glycoprotein ZP1 and its genomic organization.

The zona pellucida composed of three or four glycoproteins plays important roles in fertilization. Our previous study showed that porcine ZP1, one of the major glycoproteins of porcine zona pellucida, was divided into two components (porcine ZP4 and ZP2), and suggested it was a homologue of mouse ZP2. In this paper we report the cloning of a cDNA for porcine ZP1 and its genomic organization. The deduced amino acid sequence of porcine ZP1 shared a 54% and 63% identity with those of mouse and human ZP2, respectively. Genomic organization of porcine ZP1 was also similar to that of mouse ZP2. The transcript of porcine ZP1 gene was detected only in growing oocytes.

Expression of recombinant human zona pellucida protein 2 and its binding capacity to spermatozoa.

The human zona pellucida (ZP) is composed of three major glycoproteins: ZP1, ZP2, and ZP3. The aim of this study was to clarify the role of ZP2 by focusing on the polypeptide structure. We produced in Escherichia coli a recombinant human ZP2 protein (rec-hZP2) corresponding to amino acid sequence 1-206 of the mature protein. The final yield of rec-hZP2 protein was 80 microg/ml Luria Broth medium. After 2-h incubation of human spermatozoa with rec-hZP2 in vitro, an immunofluorescent study indicated that rec-hZP2 bound only to acrosome-reacted spermatozoa. The binding site migrated from the acrosome to the midpiece of the spermatozoa. Rabbit and mouse antisera produced against rec-hZP2 stained native human ZP in the immunofluorescent study, and significantly blocked human sperm binding and penetration into human ZP as compared to control values. The N-terminal polypeptide portion of human ZP2 was shown to contain a binding site for acrosome-reacted spermatozoa and to play an important role in secondary sperm binding and penetration into the ZP.

Epitope analysis for human sperm-immobilizing monoclonal antibodies, MAb H6-3C4, 1G12 and campath-1.

Human monoclonal antibody, MAb H6-3C4, possesses strong sperm immobilizing activity. MAb H6-3C4 has been suggested by several research groups to react with a carbohydrate moiety of male reproductive tract CD52 (mrtCD52). In the present study, we analysed the epitope on mrtCD52 for MAb H6-3C4 and found that it was polymorphic in Western blot analysis and disappeared after enzymatic removal of the N-linked carbohydrate moiety. Two other monoclonal antibodies (1G12, campath-1) with sperm-immobilizing activity recognized mrtCD52 in a polymorphic manner similar to MAb H6-3C4. Further analysis showed that 1G12 recognized a structure formed by the peptide and/or a glycosylphosphatidylinositol (GPI) anchor portion as does campath-1. Results of a lectin binding assay suggested the presence of O-linked carbohydrates on mrtCD52. Our results also indicated that the peptide portion of CD52 could serve as an epitope for sperm-immobilizing antibodies. It was concluded that the epitope of MAb H6-3C4 is similar to, but distinct from, those of 1G12 and campath-1, and that mrtCD52 contains different antigenic epitopes.

Studies on the epitope of pig zona pellucida recognized by a fertilization-blocking monoclonal antibody.

Heat solubilized pig zona pellucida (pZP) was separated into four major glycoprotein families (pZP1, pZP2, pZP3, pZP4). Cloning of a full-length cDNA coding for pZP1 revealed that the coding sequence for pZP4 was immediately followed by the sequence for the amino-terminal residue of pZP2 and that they shared a significant similarity with mouse and human ZP2, which are considered to serve as a secondary sperm receptor in sperm-zona interactions. One of the monoclonal antibodies, mAb-5H4, produced against pZP4 inhibited the binding of boar and human spermatozoa to each of the homologous oocytes. The epitope recognized by mAb-5H4 was determined to be present on a 10 amino acid sequence of pZP1(50-59) by using epitope mapping and analysis of chain flexibility of pZP4. A synthetic 18mer peptide corresponding to pZP1(50-67) reacted with mAb-5H4, and mouse antisera raised to the synthetic 18mer peptide recognized intact pig zona pellucida and strongly inhibited the fertilization of pig oocytes with boar spermatozoa in vitro. These results suggest that the pZP4 peptide (50-59) identified as an epitope for mAb-5H4 could be a promising candidate in the development of a contraceptive vaccine.

Genomic organization of the gene for pig zona pellucida glycoprotein ZP1 and its expression in mammalian cells.

The pig zona pellucida (pZP) is composed of glycoproteins pZP1, pZP3 alpha and pZP3 beta. Previously, we showed that pZP1 was proteolytically cleaved into two components (pZP4 and pZP2), and suggested that pZP1 was a homologue of mouse ZP2. We now report the genomic organization of pZP1 gene, and expression of the recombinant protein in mammalian cells. Two genomic clones, pZP4g and pZP4.1, were isolated by screening the pig genomic library with an already cloned cDNA encoding pZP4 as a probe. These clones covered 25 kb of the genome, including the entire pZP1 locus. The pig ZP1 genome consists of 18 exons and 17 introns. For production of the recombinant proteins of pZP1, a minigene containing a genomic fragment covering exon 6 to exon 17 was constructed and cloned into the expression vector, pBCMGS-neo. Mammalian cell lines, CHO (for stable expression) and 293T (for transient expression), transfected with the plasmid construct synthesized pZP1 recombinant proteins, probably as a membrane-bound protein.

Binding of recombinant pig zona pellucida protein 1 (ZP1) to acrosome-reacted spermatozoa.

In an immunofluorescent study, recombinant pig zona pellucida protein 1 (pZP1) produced in Escherichia coli bound to acrosome-reacted spermatozoa from five different mammals. The location of the binding site moved from the equatorial region to the midpiece and tail regions during the incubation. This finding suggests that pig ZP1 acts as a secondary sperm-binding receptor in an interspecies manner and assists penetration of the spermatozoa through the zona pellucida. The recombinant pZP1 bound to 55 kDa protein (proacrosin) and 40 kDa protein of the boar spermatozoa as revealed by immunoblotting.