A promising poly (e-caprolactone)/graphene-based scaffold as an antibacterial in regenerating bone tissue.

Scaffolds are 3D biomaterials that provide an environment for cell regeneration. In the context of bone remodeling, poly(e-caprolactone) (PCL) combined with graphene has been developed as the scaffold. It is imperative for scaffolds to possess antibacterial properties in order to properly reduce the risk of potential infections.Therefore, this study aims to analyze the antibacterial characteristics of PCL/graphene scaffolds against Staphylococcus aureus (S. aureus) and Porphyromonas gingivalis (P. gingivalis) in vitro. In this study, five different groups were used, including PCL (K-), Amoxicillin (K+), PCL/Graphene 0.5 wt%, PCL/graphene 1 wt% and PCL/Graphene 1.5 wt%. All experiments were performed in triplicates and were repeated three times, and the diffusion method by Kirby-Bauer test was used. The disc was incubated with S. aureus and P. gingivalis for 24 hours and then the diameter of the inhibition zone was measured. The results showed that the PCL/graphene scaffolds exhibited dose-dependent antibacterial activity against S. aureus and P. gingivalis. The inhibition zone diameter (IZD) against S. aureus of PCL/graphene 1 wt% was 9.53 ± 0.74 mm, and increased to 11.93 ± 0.92 mm at a concentration of 1.5 wt% of graphene. The PCL/graphene scaffold with 1.5 wt% exhibited a greater inhibitory effect, with an IZD of 12.56 ± 0.06 mm against P. gingivalis, while the inhibitory activity of the 1 wt% variant was relatively lower at 10.46 ± 0.24 mm. The negative control, PCL, and PCL/graphene 0.5 wt% exhibited no antibacterial activity sequentially (p = 1). Scaffolds of poly(e-caprolactone)/graphene exhibited an antibacterial activity at 1, and 1.5 wt% on S. aureus and P. gingivalis. The antibacterial properties of this scaffold make it a promising candidate for regenerating bone tissue.

Loss of PTEN expression is an independent predictor of favourable survival in endometrial carcinomas.

BACKGROUND: We and others previously reported the prognostic significance of PTEN mutational status on favourable survival in endometrial carcinomas. Here, we demonstrate that loss of PTEN expression in immunohistochemistry is an independent prognostic marker for favourable survival in endometrial carcinomas. METHODS: We conducted immunohistochemical analyses of PTEN, PIK3CA, phosphorylated Akt (p-Akt), and p27 in primary endometrial carcinomas from 221 patients. Mutation of PTEN was analysed further. RESULTS: Expression of PTEN was lost in 56 patients (25%), and PIK3CA was overexpressed in 159 patients (72%). Overexpression of PIK3CA was associated with p-Akt overexpression (P<0.001), which was in turn associated with loss of nuclear p27 expression (P=0.028). Loss of PTEN expression was found to be associated with endometrioid histology (P=0.03), and was inversely associated with the presence of lymphovascular space invasion (P=0.03). Univariate and multivariate survival analyses revealed that factors of PTEN loss, age <70, histological grade 1, early International Federation of Gynecology and Obstetrics (FIGO) stage, and absence of lymphovascular invasion were independent prognostic indicators for better overall survival (P=0.03, 0.04, 0.01, <0.001, and 0.03, respectively). The subset analysis showed a stronger tendency of PTEN loss towards favourable survival in advanced-stage (III and IV) disease than in early-stage (I and II) disease (P=0.05 vs 0.14). Moreover, our mutational analysis demonstrated that PTEN expression loss was associated with PTEN-truncating mutations (P=0.03). CONCLUSION: The current observations further support the prognostic significance of PTEN aberration on favourable outcome in endometrial carcinomas, providing useful implications for the individualised management of the disease.

Limit on neutrinoless ßß decay of 136Xe from the first phase of KamLAND-Zen and comparison with the positive claim in 76Ge.

We present results from the first phase of the KamLAND-Zen double-beta decay experiment, corresponding to an exposure of 89.5 kg yr of (136)Xe. We obtain a lower limit for the neutrinoless double-beta decay half-life of T(1/2)(0ν)>1.9×10(25) yr at 90% C.L. The combined results from KamLAND-Zen and EXO-200 give T(1/2)(0ν)>3.4×10(25) yr at 90% C.L., which corresponds to a Majorana neutrino mass limit of <(120-250) meV based on a representative range of available matrix element calculations. Using those calculations, this result excludes the Majorana neutrino mass range expected from the neutrinoless double-beta decay detection claim in (76)Ge, reported by a part of the Heidelberg-Moscow Collaboration, at more than 97.5% C.L.

Exploiting multimedia in reproductive science education: research findings.

Education in reproductive science is operating from an outdated paradigm of teaching and learning. Traditionally, reproductive education follows the pattern where students read a textbook, listen to instructor presentations, re-read the textbook and class notes and then complete a test. This paradigm is inefficient, costly and has not incorporated the potential that technology can offer with respect to increases in student learning. Further, teachers of reproductive science (and all of science for that matter) have little training in the use of documented methods of instructional design and cognitive psychology. Thus, most of us have learned to teach by repeating the approaches our mentors used (both good and bad). The technology now exists to explain complex topics using multimedia presentations in which digital animation and three-dimensional anatomical reconstructions greatly reduce time required for delivery while at the same time improving student understanding. With funding from the Small Business Innovation Research program through the U.S. Department of Education, we have developed and tested a multimedia approach to teaching complex concepts in reproductive physiology. The results of five separate experiments involving 1058 university students and 122 patients in an OB/GYN clinic indicate that students and patients learned as much or more in less time when viewing the multimedia presentations when compared to traditional teaching methodologies.

T cell tolerance studied at the level of antigenic determinants. I. Latent reactivity to lysozyme peptides that lack suppressogenic epitopes can be revealed in lysozyme-tolerant mice.

Whether T cell tolerance represents direct inactivation of antigen-specific T cells via recognition of antigen plus major histocompatibility complex, or via T suppressor (Ts) cells, or a combination of these mechanisms, remains to be clarified. This problem was investigated using a novel approach based on the finding in several systems that T helper/proliferative (Th/Tp) cell-inducing antigenic determinants are dissociable from Ts cell-inducing determinants. Thus, peptide probes containing known sites that stimulate T proliferative activity, as well as peptides from distinct sites assumed to bear Ts-inducing determinants, were used in studying hen (chicken) eggwhite lysozyme (HEL)-tolerant mice. The clear prediction from clonal deletion model is that Th/Tp response potential to short peptides in the tolerant mouse would not exist, while regulatory suppression models predict the coexistence of antigen-reactive cells and antigen-specific regulatory cells that prevent their expression. Adult mice, treated with 2 mg HEL in saline, were tolerant to HEL in complete Freund's adjuvant (CFA). Latent T cell proliferative responses could be revealed to determinants within two HEL peptide probes, which lacked the amino-terminal region of the molecule. This responsiveness suggested two conclusions: first, Ts cells directed against the amino terminus of lysozyme exist in the tolerant genetic responder B10.A; second, these Ts regulate the activity of functional antigen-reactive T cells directed against epitopes elsewhere on the molecule, but only in the presence of the complete molecule, HEL. Examination of neonatally induced tolerance did not reveal any latent responsiveness, supporting the hypothesis that clonal deletion or anergy is the relevant mechanism in this situation. Possible reservations in these explanations of the two tolerant states, plus analysis of the more complex "split tolerance" resulting from 20 mg HEL in saline treatment in adults, are discussed. The approach of dissociation of proliferation-inducing determinants from suppression-inducing determinants clarifies our understanding of the tolerant state and holds promise for more definitive exploration of mechanisms of T cell tolerance.

Induction of tolerance to one determinant on a synthetic peptide does not affect the response to a second linked determinant. Implications for the mechanism of neonatal tolerance induction.

To investigate the mechanism underlying neonatal T cell tolerance, we used synthetic peptides to induce tolerance. We found that induction of tolerance to one determinant on a 23-amino acid peptide did not affect the response to an adjacent determinant on the same peptide. There was no evidence of suppression of the response to the second determinant. Furthermore, even small peptides near the minimal size for a determinant, which would be very unlikely to possess a suppressor T cell-inducing determinant as well as a proliferative T cell-inducing determinant, could induce tolerance. These studies provide in vivo experiments supporting clonal inactivation as the mechanism of neonatal tolerance to immunogenic peptides.

Distinct recognition phenotypes exist for T cell clones specific for small peptide regions of proteins. Implications for the mechanisms underlying major histocompatibility complex-restricted antigen recognition and clonal deletion models of immune response gene defects.

Using synthetic peptides as antigens, it was found that T cell clones of a given haplotype specific for 13-16 amino acid peptides could be clearly distinguished by the varied influence of amino acid substitutions on recognition. This was true for different antigenic determinants within peptides 81-96 and 74-86 of hen egg-white lysozyme, recognized in the context of the I-Ab and I-Ak molecules, respectively. Considerable complexity was demonstrated in the induced T cell repertoire specific for apparently single determinants, which implies that diversity of T cell recognition approaches that for B cells. The implications of the degeneracy of T cell recognition are discussed in the context of mechanisms through which Ia molecules restrict recognition and theories of Ir gene defects.

Silent venous thromboembolism before treatment in endometrial cancer and the risk factors.

Venous thromboembolism (VTE) often occurs after surgery and can even occur before surgery in patients with gynaecological malignancies. We investigated the incidence of VTE before treatment of endometrial cancer and associated risk factors. Plasma D-dimer (DD) levels before initial treatment were examined in 171 consecutive patients with endometrial cancer. Venous ultrasound imaging (VUI) of the lower extremities was performed in patients with DD >or=1.5 microg ml(-1), as the negative predictive value of DD for VTE is extremely high. For patients with deep vein thrombosis (DVT), pulmonary scintigraphy was performed to ascertain the presence of pulmonary thromboembolism (PTE). Risk factors for VTE were analysed using univariate and multivariate analyses for 171 patients. Of these, 37 patients (21.6%) showed DD >or=1.5 microg ml(-1), 17 (9.9%) displayed DVT by VUI and 8 (4.7%) showed PTE on pulmonary scintigraphy. All patients with VTE were asymptomatic. Univariate analysis for various risk factors revealed older age, non-endometrioid histology and several variables of advanced disease as significantly associated with VTE before treatment. Obesity, smoking and diabetes mellitus were not risk factors. Multivariate analysis confirmed extrauterine spread and non-endometrioid histology as independently and significantly associated with risk of VTE. These data suggest that silent or subclinical VTE occurs before treatment in at least around 10% of patients with endometrial cancer. Risk factors for VTE before treatment might not be identical to those after starting treatment.

Alleles and intron polymorphism of KIR3DL1 shown by combination of allele group-specific primers and sequencing.

Allelic polymorphism of killer immunoglobulin-like receptor (KIR) genes may affect receptor expression as well as function. We used a combination of allele group-specific primers and DNA sequencing techniques to verify our KIR genotyping primers in polymerase chain reaction and identified three KIR3DL1 alleles. By sequencing some introns of 3DL1 in 18 genomic DNA samples, we found that a 4-bp insert in intron 1 of 3DL1*002 exists in multiple alleles, but that a 2-bp deletion in intron 7 is unique to 3DL1*01501 and that a 19-bp insert in intron 1 seems specific to the CEPH family 1416. Our data suggest that extensive KIR gene polymorphisms are ubiquitous as well as quite complex.

Prevalence and clinical features of diabetes mellitus secondary to chronic pancreatitis in Japan; a study by questionnaire.

The prevalence and clinical features of diagnosed mellitus secondary to chronic pancreatitis (CP) were assessed from northern (Hokkaido) to southern (Okinawa) Japan by means of a questionnaire to elucidate whether WHO-classified malnutrition-related diabetes mellitus (MRDM) exists in Japan. Of a total 17,500 diabetic patients, only two (0.011%)-one fibrocalculous pancreatic diabetes (FCPD) and one protein-deficient pancreatic diabetes (PDPD) - exhibited MRDM characteristics. A total of 649 CP were collected and classified into 268 cases with chronic alcoholic pancreatitis (CAP), 150 cases with chronic calcified pancreatitis (CCP) and 231 cases with other CP. The prevalence of diabetes mellitus was found to be 50.7% in CAP, 72.7% in CCP and 22.8% in other CP. Among all diabetics, 56.6% was noninsulin-dependent (NIDDM) and 26.4% insulin-dependent (IDDM). IDDM was most frequent in CP. Satisfactory and less than satisfactory glycemic control was obtained in approximately three quarters of all subjects. Only one quarter showed poor glycemic control. Insulin treatment was frequent in CAP (52.2%) and CCP (61.7%), but less in other CP (27.5%). The prevalence of diabetic retinopathy was observed in 33.1% of all subjects, nephropathy 21.0% and neuropathy 36.3%, respectively. The prevalence of complications, including macroangiopathy tended to be higher in CAP and CCP (40.3 and 56.9%) than in other CP (31.4%).

A novel human glassy-cell carcinoma cell line producing IL-6 and IL-8 from uterine cervix.

A novel human cell line, TOM-2, was established from a rare uterine cervical cancer, glassy cell carcinoma (GCC). TOM-2 is the second established GCC cell line so far reported. The cells were intermediately or poorly differentiated with dysplastic nuclei and polygonal shape and secreted two tumor markers and cytokines, i.e., CA-125 and SCC, interleukin (1L)-1alpha, -6, and -8, and TNF-alpha. Growth of TOM-2 was so strongly dependent on population density that it was not possible to determine the plating efficiency. In mass culture, the following characteristics were observed: doubling time, 83 h; mode of chromosome number, 79; human papillomavirus type 18 DNA, detectable; tumorigenicity, easily transplantable into subcutis of nude mice; chemosensitivity in vitro, considerably sensitive to Cisplatin and 5-FU but not to 9 other antineoplastic agents. This novel cell line will be useful for developing new therapeutic strategies for the rare cancer, GCC.

[Establishment of a new human endometrial adenocarcinoma cell line, Watanabe cells, containing estrogen receptor].

A new human endometrial adenocarcinoma cell line, Watanabe cells, was established from the ascitic fluid of a relapsed endometrial adenocarcinoma obtained from a 58-year-old woman; this cell line has been maintained in vitro for more than 3 years and 8 months. The cells formed a monolayer in a mosaic fashion and tended to pile up and formed a hemicyst. The population boubling time was 60.0 hours at the 10th generation. The modal chromosomal number of the cells was in the diploid range. The histology of tumors induced by this cell line in athymic nude mice showed poorly differentiated adenocarcinoma, while the initial tumor was a well differentiated adenocarcinoma. Estrogen and progesterone receptors (ER, PR) were demonstrated in the original tumor, whereas ER but not PR were present in the tumors induced in nude mice. CA125, CA19-9 and other tumor markers were positive in culture media of this cell line. The cells showed intrinsic cisplatin-resistance (50% inhibition concentration: > 10 micrograms/ml) at 120 hours of exposure by MTT assay. We believe this cell line will be useful for investigating the mechanisms of progesterone therapy, the biological behaviors of the tumor markers and mechanisms of chemotherapeutic resistance in endometrial carcinoma.

Establishment of eighteen clones of Ishikawa cells.

Ishikawa cells, which derive from a well differentiated human endometrial adenocarcinoma, were cloned using the limiting dilution method in an attempt to preserve well differentiated cells. Eighteen clones were obtained. According to their morphologic characteristics, there were six well differentiated, seven moderately differentiated, three poorly differentiated and two adenosquamous carcinoma clones. All of them were transplantable into node mice. Fifteen of the 17 clones were positive for estrogen receptors and all of the clones examined were positive for progesterone receptors. There were no significant differences in terms of population doubling time, plating efficiency or saturation density among these clones. On the basis of these results, we concluded that Ishikawa cells are not as homogeneous morphologically as we thought, since cells with varying degrees of differentiation have already mingled with the parent cell line. It is verified that the adenosquamous cell carcinoma of the endometrium can arise from one stem cell. It would be impossible to revive the cultured cells after having undergone their changes to the original conditions. Therefore, to preserve the characteristics of an established cell line, the cells should be frozen at a very early stage. It is also important to avoid frequent passages when special characters of the cells are necessary for a given investigation.

[Establishment of a cisplatin-resistant new human endometrial adenocarcinoma cell line, Sawano cells].

A new human endometrial adenocarcinoma cell line, Sawano cells, was established from an endometrial adenocarcinoma from a 61-year-old woman and has been maintained in vitro for more than 2 years and 10 months. The cells were found to form a monolayer in a mosaic fashion and to tend to pile up. Population doubling time was 43.2 hours at the 10th generation. The modal chromosomal number of the cell fell in a diploid range. Histology of the tumor induced in athymic mice showed it to be a moderately differentiated adenocarcinoma which closely resembled the original human tumor. Estrogen and progesterone receptors were not demonstrated in the in vitro culture cells and in the tumors induced in nude mice, though they were positively demonstrated in the original tumor. The cells had intrinsic cisplatin-resistance (50% inhibition concentration:6.63 micrograms/ml) at 120 hours of contact. We believe this cell line with help us to investigate the biological mechanisms of CDDP resistance.

IGF-II producing hepatic fibrosarcoma associated with hypoglycemia.

A 67-year-old male was admitted with the complaint of weakness at hunger early in the morning, when blood glucose was less than 40 mg/dl. The abdominal ultrasonogram and computerized tomogram demonstrated a huge tumor in the right liver lobe. Hypoglycemia disappeared after transcatheter arterial embolization. Then hepatic lobectomy was performed. The tumor was histologically shown to be a fibrosarcoma. Insulin-like growth factor-II was intensely stained in the Golgi area of the tumor cells, suggesting its role in the mechanism of hypoglycemia.

Five cases of asymptomatic spontaneous pneumothorax.

Asymptomatic spontaneous pneumothorax (ASPT) is an uncommon condition. Between January 1, 1989 and December 31, 1997, 269 patients were admitted to our department with spontaneous pneumothorax. Of the 269 patients, 5 had no symptoms at the time of discovery. Their ages ranged from 15 to 61 years (mean, 37.8 years), and all of them were male. Of the 5 patients with no complaints, 2 had bilateral metachronous pneumothoraces and 3 had hemilateral pneumothorax. All of these ASPTs were revealed by chest roentgenographs taken during medical examinations or follow-up studies relating to other diseases. The mean value of body mass index (BMI) was 19.96 +/- 1.4 (range 18.7 - 22.1). Two of the 5 patients underwent bilateral partial lung resection. Histopathological examination of the resected specimens showed elastofibrosis, scar formation, and an interruption of the elastic fiber of the pleura. In these 5 cases, clinical courses were uneventful, and relapse of the pneumothorax did not occur. Clinical physicians should be aware of the possibility of asymptomatic pneumothorax, as well as the optimal radiographic techniques for revealing small pneumothoraces.

[A pharmacokinetic study of cefoperazone during percutaneous transhepatic cholangial catheterization].

The metabolic fate of cefoperazone (CPZ) was studied in 19 cases which underwent percutaneous transhepatic cholangial catheterization (PTC-catheterization, PTCC) and were under various conditions of the liver function. The peak of bile levels of CPZ immediately after PTCC differed greatly from one case to another at 12.6-7,260 micrograms/ml with 1 g intravenous injection and 23.0-5,800 micrograms/ml with 2 g intravenous injection. The ratio of the peak of bile level to the serum level immediately after PTCC showed the highest negative correlation with the serum total bilirubin level. It also showed a significant negative correlation with GOT, GPT, Al-P and LAP. The serum CPZ level and half-life showed no significant trend except half-life showed a significant correlation with LAP. The recovery rate in urine up to 12 hours was in the range of 14.8-93.6%, showing a significant correlation with the ratio of the peak of bile levels to the serum level and the date of liver function tests. The bile level, serum level and recovery rate in urine at the time the bile outflow from the catheter has become constant after PTCC (during the course of PTCC) showed a trend almost similar to that immediately after PTCC, there being no significant difference as to each parameter during the course of PTCC and immediately after PTCC. In the cases in which the sample was collected by the cross-over technique, the ratio of the peak of bile levels to the serum level from immediately after PTCC to during the course of PTCC increased in 2 cases and decreased in 6 cases. The 2 cases that showed the increase in the ratio were the case in which the serum total bilirubin level improved almost to normal. Findings above suggest that sufficient biliary decompression can improve the movement of CPZ into bile, despite the fact that the pharmacokinetics of CPZ is affected by the liver function, particularly serum total bilirubin level, that a decrease in the movement to bile and a compensatory increase in urinary excretion are observed in jaundice and disturbance of the liver function and that the ratio of the peak of bile level to the serum level decreases during the course of PTCC rather than immediately after PTCC in some cases.

[Surgically treated pancoast tumor].

A 40-year-old male was detected his right apical lung tumor by roentgenographic screening on January 1997, but he did not refer to a hospital since he had no symptom. He went a orthopedics because of his right chest, back, and arm pain on October 1997, and he received traction and physical therapy. He went roentgenographic screening again on January 1998 and he was pointed out that the tumor increased. He admitted our hospital. Biopsy using bronchofiberscopy revealed adenocarcinoma and induction radiotherapy (40 Gy) was performed. Right upper lobectomy with chest wall resection and lymph node dissection was performed under hook approach. This approach was useful to dissect the tumor from the invaded plexus brachialis. Postoperative radio-chemotherapy was added but the patient died 7-postoperative months because of multiple metastases. Early detection should be led to early starting of the therapy.