Outcome of fertility-sparing treatment with medroxyprogesterone acetate for atypical hyperplasia and endometrial carcinoma in young Japanese women.

PURPOSE: To review the outcome in patients with atypical endometrial hyperplasia (AEH) and endometrial cancer (EC) who received MPA treatment in the present hospital. MATERIALS AND METHODS: Patients with AEH or EC were administered MPA for 12 weeks followed by endometrial curettage. The rates of effect, recurrence, pregnancy, and complications were evaluated. The changes in progesterone receptors and FOXO-1, known as a target of MPA treatment, were examined by immunostaining. RESULTS: Four of seven patients with endometrial cancer and three of three patients with AH had complete response. Four of seven patients had recurred within one year after the treatment and had to undergo hysterectomy. None of the patients showed changes in progesterone receptors. Although six of seven patients were negative for FOXO-1 before and after treatment, all the patients showed increased developments of FOXO-1 during MPA treatment. CONCLUSION: Progestin as a fertility-preserving treatment is expected to be effective for endometrial cancer, but judicious use might be required because it shows high rate of recurrence. Further studies regarding the mechanism may be necessary to achieve high efficacy.

Evaluation of primary prophylaxis with granulocyte colony-stimulating factor for epithelial ovarian cancer.

PURPOSE: Primary prophylaxis with G-CSF has been used to minimize myelosuppression caused by anticancer agents and to avoid severe neutropenia. The authors retrospectively examined the value of primary prophylaxis using granulocyte colony-stimulating factor (G-CSF) for epithelial ovarian cancer. MATERIALS AND METHODS: From 2001 to 2010, 105 patients with ovarian cancer receiving chemotherapy in the present hospital were divided into two groups: one received primary prophylaxis with G-CSF and the other did not receive it in compliance with the guidelines for G-CSF usage. The incidence of febrile neutropenia (FN), degree of neutropenia, frequency of G-CSF administration, number of days of hospitalization, progression-free survival (PFS), and overall survival (OS) were evaluated. RESULTS: Neutrophils decreased almost equally and the length of hospitalization was not significantly lower between the groups. Five-year PFS or OS showed no significant difference either. CONCLUSIONS: Primary prophylaxis with G-CSF in chemotherapy for epithelial ovarian cancer could be of low significance.

New candidates for mechano-sensitive channels potentially involved in gravity sensing in Arabidopsis thaliana.

The mechano-sensitive channels of plants may sense increases in tension induced by mechanical stimuli, such as touch, wind and turgor pressure, and a gravitational stimulus. Recent studies have identified plant homologues of the bacterial mechano-sensitive channel MscS, which is gated by membrane tension and reduces intracellular osmolality by releasing small osmolytes from bacterial cells. However, the physiological roles of these homologues have not yet been clearly elucidated, and only two of them have been shown to be involved in the protection of osmotically stressed plastids in Arabidopsis thaliana. We identified another group of candidates for mechano-sensitive channels in Arabidopsis, named MCA1 and MCA2, whose homologues are exclusively found in plant genomes. MCA1 and MCA2 are composed of 421 and 416 amino acid residues, respectively, share 73% homology in their amino acid sequences, and are not homologous to any known ion channels or transporters. Our structural study revealed that the N-terminal region (one to 173 amino acids) of both proteins was necessary and sufficient for Ca(2+) influx activity. Interestingly, this region had one putative transmembrane segment containing an Asp residue whose substitution mutation abolished this activity. Our physiological study suggested that MCA1 expressed at the root tip was required for sensing the hardness of the agar medium or soil. In addition, MCA1 and MCA2 were shown to be responsible for hypo-osmotic shock-induced increases in [Ca(2+) ]cyt . Thus, both proteins appear to be involved in the process of sensing mechanical stresses. We discussed the possible role of both proteins in sensing mechanical and gravitational stimuli.

Mechanosensitive channels are activated by stress in the actin stress fibres, and could be involved in gravity sensing in plants.

Mechanosensitive (MS) channels are expressed in a variety of cells. The molecular and biophysical mechanism involved in the regulation of MS channel activities is a central interest in basic biology. MS channels are thought to play crucial roles in gravity sensing in plant cells. To date, two mechanisms have been proposed for MS channel activation. One is that tension development in the lipid bilayer directly activates MS channels. The second mechanism proposes that the cytoskeleton is involved in the channel activation, because MS channel activities are modulated by pharmacological treatments that affect the cytoskeleton. We tested whether tension in the cytoskeleton activates MS channels. Mammalian endothelial cells were microinjected with phalloidin-conjugated beads, which bound to stress fibres, and a traction force to the actin cytoskeleton was applied by dragging the beads with optical tweezers. MS channels were activated when the force was applied, demonstrating that a sub-pN force to the actin filaments activates a single MS channel. Plants may use a similar molecular mechanism in gravity sensing, since the cytoplasmic Ca(2+) concentration increase induced by changes in the gravity vector was attenuated by potential MS channel inhibitors, and by actin-disrupting drugs. These results support the idea that the tension increase in actin filaments by gravity-dependent sedimentation of amyloplasts activates MS Ca(2+) -permeable channels, which can be the molecular mechanism of a Ca(2+) concentration increase through gravistimulation. We review recent progress in the study of tension sensing by actin filaments and MS channels using advanced biophysical methods, and discuss their possible roles in gravisensing.

Effect of ageing on healing of bilateral mandibular condyle fractures in a rat model.

To examine the hypothesis that conservative treatment is applicable to younger patients with bilateral mandibular condylar fractures, we studied the effect of ageing on the healing of bilateral mandibular condylar fractures in a rat model. Male Sprague-Dawley rats aged 3, 6, and 36 weeks (n=25/cohort, total n=75) were divided into a fracture group (n=12) and a sham control group (n=12); one rat from each cohort was used as a normal unoperated control. Cell proliferation was evaluated using the bromodeoxyuridine (BrdU) labelling index (LI). Osteochondrogenesis was assessed by the expression of Indian hedgehog (Ihh), type X collagen, and osteocalcin in the condylar head. Condylar fracture healing was found to be delayed by ageing. BrdU LI values in the fracture groups were higher in younger rats than in older rats at 8 weeks after fracture. The number of Ihh-positive cells in the fracture groups increased significantly up to 2 weeks after fracture, and then gradually decreased until 8 weeks after fracture. The findings of this study support the clinical concept of conservative treatment of bilateral condylar fractures in younger patients, but functional issues regarding ramus height and its consequences on occlusion have not been tested in this study.

The retromandibular transparotid approach for reduction and rigid internal fixation using two locking miniplates in mandibular condylar neck fractures.

We evaluated the safety, efficacy, and morbidity associated with the treatment of displaced mandibular condylar neck fractures using a retromandibular transparotid approach to reduce and rigidly fix using two 2.0-mm locking miniplates. Our surgical inclusion criteria were: patient selection of open reduction and fixation, displaced unilateral condylar fractures with derangement of occlusion, and bilateral condylar fractures with an anterior open bite. The study group consisted of 19 patients who underwent surgery for 19 mandibular condylar neck fractures; patients were analyzed prospectively, with more than 6 months of follow-up, and were evaluated in terms of functional results, scar formation, postoperative complications, and stability of fixation. The results showed that functional occlusion identical to the preoperative condition and correct anatomical reduction of the condylar segments in centric occlusion, followed by immediate functional recovery, was achieved in all patients. No patient suffered from any major or permanent complication postoperatively, although there were two cases (11%) of temporary facial nerve palsy, which resolved completely within 3 months. Surgical scars were barely visible. The retromandibular transparotid approach with open reduction and rigid internal fixation for displaced condylar neck fractures of the mandible is a feasible and safe, minimally invasive surgical technique that provides reliable clinical results.

Asialoglycoprotein receptor scintigraphy with 99mTc-galactosyl human serum albumin (99mTc-GSA) as an early predictor of survival in acute liver failure.

This study evaluated the usefulness of asialoglycoprotein receptor scintigraphy with 99mTc-galactosyl human serum albumin (99mTc-GSA scintigraphy) as an early predictor for prognosis of acute liver failure. Forty-eight patients with acute liver failure and without a past history of chronic liver disease were enrolled. Patients were divided into survival and non-survival groups by 28-day mortality. 99mTc-GSA scintigraphy to detect uptake ratio of the heart at 15 minutes to that at three minutes (HH15) and uptake ratio of the liver at 15 minutes to the liver plus the heart at 15 minutes (LHL15), and measurements of serum total bilirubin, hepatocyte growth factor and prothrombin time were performed immediately after the diagnosis of acute liver failure. Areas under the receiver operating characteristic curves were used to compare the prognostic ability of total bilirubin, hepatocyte growth factor, prothrombin time, HH15 ratio, LHL15 ratio and the model for end-stage liver disease score. Clinical characteristics of patients in the survival group (n=20) and in the non-survival group (n=28) were not significantly different. HH15 and LHL15 uptake ratios in the survival group were 0.670 and 0.875, and they were significantly lower and higher than those in the non-survival group, respectively. All patients with LHL15 <0.760 died, and the area under the receiver operating characteristic curve for LHL15 were significantly larger than the areas under the receiver operating characteristic curves of serum variables and model for end-stage liver disease score. In summary, in patients with acute liver failure without chronic liver disease, HH15 and LHL15 of 99mTc-GSA scintigraphy are more useful variables in predicting prognosis than serum variables and model for end-stage liver disease score.

Applicability of buccal fat pad grafting for oral reconstruction.

This study evaluated the applicability of pedicled buccal fat pad grafting for the reconstruction of defects surgically created during oral surgery. A buccal fat pad graft was applied in 23 patients (5 males, 18 females; mean age 68.3 years) between 2003 and 2011. The graft was used to cover surgical defects of the palate, maxilla, upper gingiva, buccal mucosa, lower gingiva, oral floor, and temporomandibular joint region. Size of the surgical defects ranged from 15mm×12mm to 30mm×40mm; size of the buccal fat pad ranged from 15mm×12mm to 43mm×38mm. A pedicled buccal fat pad was prepared by incising the maxillary vestibule following primary surgery, and the surrounding connective tissue was preserved to supply nutrition to the pedicle during surgery. The buccal fat pad was placed on the raw surface of soft tissue or bone surface and sutured to the surrounding tissue of the defect. Complete epithelialization was observed within 4 weeks postoperatively. There were no complications or functional disorders during follow-up. Buccal fat pad grafting appears to be feasible for the reconstruction of surgically induced defects, and can be extended to the palate, mandible, mouth angle, and temporomandibular joint region.

Lymphoepithelial-like carcinoma of the uterine cervix; a case report.

Lymphoepithelioma-like carcinoma (LELC) of the uterine cervix is a rare variant of squamous cell carcinoma of the uterine cervix. This tumor is characterized by nests of poorly differentiated epithelial cells surrounded by a prominent lymphocytic infiltration. Despite the poorly differentiated pathological findings, it appears to have a better outcome than the usual squamous cell carcinoma of the uterine cervix. Therefore, it is quite important to differentiate this tumor from poorly differentiated squamous cell carcinoma and lympho-proliferative disorders of the cervix. LELC arising from the nasopharynx has been suggested to be associated with Epstein-Barr virus (EBV), whereas the involvement of EBV in LELC of the uterine cervix is still controversial. In addition, the role of high-risk human papilloma virus (HPV) in this type of tumor remains unknown. We report a case of LELC of the cervix with diagnosis on the basis of histopathology in a 52-year-old Japanese woman who presented with a history of continuous bleeding post menopause. We also examine the association of EBV and HPV in this case.

Nominal free air in the left inguinal fossa due to perforation of the sigmoid colon in a case of blunt abdominal trauma: CT diagnosis.

We present a case of perforation of the sigmoid colon due to blunt abdominal trauma. Computed tomography showed nominal free air in the inguinal fossa. The distribution of free air may be a clue to the site of an injured intestine. Early detection of intestinal injury is difficult, but repeated computed tomography after several hours may reveal increased free air.

Improved bioluminescent enzyme immunoassay for the rapid detection of Salmonella in chicken meat samples.

AIMS: To evaluate an improved bioluminescent enzyme immunoassay (BEIA) using biotinylated firefly luciferase for the rapid detection of Salmonella in naturally contaminated chicken meat samples. METHODS AND RESULTS: Capture agents and lipopolysaccharide (LPS) extraction reagents for Salmonella were investigated to improve the sensitivity of the BEIA. Also, the use of Oxoid SPRINT (Simple Pre-enrichment and Rapid Isolation New Technology) as a pre-enrichment and selective medium for 26-h BEIA detection of Salmonella in chicken meat samples was examined. The use of polymyxin B as a capture agent on solid support and 3-[(3-Cholamidopropyl) dimethylammonio] propanesulfonic acid (CHAPS) for extraction of the LPS facilitated sensitive detection of Salmonella. Of 120 chicken meat samples, 25 samples were positive using the improved BEIA with the SPRINT and 25 samples were positive using the SPRINT followed by the standard isolation methods. CONCLUSIONS: The improved BEIA, in which polymxin B was used as a capture agent and CHAPS was used for extraction of the antigen, had a sensitivity of 96.0% and a specificity of 98.9% for the detection of Salmonella in chicken meat. SIGNIFICANCE AND IMPACT OF THE STUDY: The improved BEIA combined with the SPRINT medium for the detection of Salmonella in chicken meat samples produced comparable results to the culture methods in 26 h.

Evaluation of a 24-hour bioluminescent enzyme immunoassay for the rapid detection of Salmonella in chicken carcass rinses.

A bioluminescent enzyme immunoassay (BEIA), using Salmonella-specific monoclonal antibody M183 for capture and biotinylated monoclonal antibody M183 for detection, was developed with InteLite AB streptavidin-biotinylated firefly luciferase complex as a reporter. Salmonella cultures were preenriched in buffered peptone water with shaking for 6 h at 37 degrees C and then selectively enriched in Muller-Kauffmann tetrathionate (MKTT) broth and modified semisolid Rappaport-Vassiliadis (MSRV) medium for 16 h at 42 degrees C. After enrichment, the total test time for the BEIA was 1.5 h. The analytical sensitivity of the BEIA ranged from 6.0 x 10(2) CFU/ml to 1.2 x 10(5) CFU/ml in MKTT and from 1.4 x 10(5) to 2.3 x 10(6) CFU/ml in MSRV using six Salmonella serovars prevalent in Canada. With enrichment cultures, the BEIA detected 1 CFU of Salmonella Typhimurium and Salmonella Enteritidis in 25 ml of chicken rinses. Representative strains of 10 Salmonella serovars were detected, and cross-reactivity was not observed with 25 non-Salmonella foodborne bacteria. The BEIA performance was assessed by testing 420 poultry samples, which were analyzed in parallel with the standard MSRV culture method. The BEIA detected 117 (27.88%) Salmonella-positive samples, whereas the standard MSRV culture method detected 124 (29.5%). The BEIA had a sensitivity of 64.5% and a specificity of 87.5% compared to the standard MSRV culture method. However, similar specificities and sensitivities were obtained when the standard MSRV culture method was compared to the BEIA (sensitivity = 68.4% and specificity = 85.5%). Neither method detected 100% of the Salmonella found in the samples tested, and statistical analyses indicated no significant difference between the two methods. In summary, the BEIA offers another alternative for the detection of Salmonella, with the additional advantage of providing a 24-h test for detecting Salmonella in chicken carcass rinses. The results obtained in this research indicate that tests are still needed for the isolation and detection of Salmonella that will establish the true prevalence of Salmonella in chicken samples.

In vitro reconstitution of signal transmission from a hair cell to the growth cone of a chick vestibular ganglion cell.

Signal transmission from a chick hair cell to the growth cone of a vestibular ganglion cell was examined by placing an acutely dissociated hair cell on the growth cone of a cultured vestibular ganglion cell. Electrical stimuli were applied to the hair cell while monitoring the intracellular Ca(2+) concentration ([Ca(2+)](i)) at the growth cone or recording whole-cell currents from the vestibular ganglion cell. Electrical stimulation of the hair cell induced [Ca(2+)](i) increases at the growth cone and inward currents in the vestibular ganglion cell. The [Ca(2+)](i) increase was blocked by 6-cyano-7-nitroquinoxaline (CNQX) (10 microM) but not by 2-amino-5-phosphonovaleric acid (APV; 50 microM). Glutamate (100 nM-300 microM) applied to the vestibular ganglion cell by the Y-tube method induced inward currents which were also antagonized by CNQX, but not by APV. These results indicate that the electrical stimulation of a hair cell induced glutamate or glutamate like agent release from the hair cell, which activated non-N-methyl-D-aspartate receptors at the growth cone of the vestibular ganglion cell, followed by action potentials and [Ca(2+)](i) elevation in the vestibular ganglion cell. This is the first demonstration of in vitro reconstitution of functional signal transmission from a hair cell to a vestibular ganglion cell.

Internet medical usage in Japan: current situation and issues.

Internet use by physicians and patients has become very popular in Japan. Fifty percent of physicians use the Internet to search for medical and other information. Over the past year, 22% of patients used the Internet to obtain medical information. Because there are no restrictions within Japan on using Web sites to advertise medical treatment, information can be freely sent out, and over the past two or three years this practice has increased dramatically. Internet medical information provides information about illnesses and medications, and it helps improve the quality of life of patients and families. Yet, depending on the content of the information provided and the way this information is used, there is a potential negative side as well. On principle, users are responsible for the way information is used, but there is a need for information providers to consider users safety and to make the information effective for use. Because there is no absolute standard for evaluating the value of medical information, it is necessary to establish a system that opens a dialogue with society and that continuously accumulates high-quality information through the collection of various evaluations, rather than rely on an established authority. For industries and organizations related to commercial pursuits, in particular, it is most effective to establish their own codes for ethical conduct, rather than rely on governmental regulations. At the same time, it is important to have a confirmation function to evaluate how goals set by the outside are being implemented. Aiming at establishing a framework for the Internet medical usage, the Japan Internet Medical Association (JIMA) was founded in 1998 by medical professionals, lawyers, researchers, consumer representatives, patients and their families. We propose a system that would combine feedback from users, who would take on the role of evaluators of the implementation of an ethical code, with a displayed mark that verifies the identity of the Web site. Objective evaluation of information is needed to ensure that users have the power to make choices. Medical experts or patient and family groups would assist in this task. The development of medical care will be promoted through patients and physicians working together in the accumulation of shared resources for good medical care information.

Dynamics of integrin clustering at focal contacts of endothelial cells studied by multimode imaging microscopy.

Human umbilical vein endothelial cells were stained with FITC-labeled anti-beta(1) integrin antibody and plated on a glass cover slip to elucidate the mechanism of integrin clustering during focal contact formation. The process of integrin clustering was observed by time-lapse total-internal-reflection fluorescence microscopy, which can selectively visualize the labeled integrins at the basal surface of living cells. The clustering of integrins at focal contacts started at 1 hour after plating and individual clusters kept growing for approximately 6 hours. Most integrin clusters (approximately 80%) elongated towards the cell center or along the cell margin at a rate of 0.29+/-0.24 microm minute(-1). Photobleaching and recovery experiments with evanescent illumination revealed that the integrins at the extending tip of the clusters were supplied from the intracellular space. Simultaneous time-lapse imaging of exocytosis of integrin-containing vesicles and elongating focal contacts showed that most exocytosis occurred at or near the focal contacts followed by their elongation. Double staining of F-actins and integrins demonstrated that stress fibers were located near the integrin clusters and that intracellular punctate integrins were associated with these stress fibers. These results suggest that the clustering of integrins is mediated by actin-fiber-dependent translocation of integrins to the extending tip of focal contacts.

Calcium-sensitive nonselective cation channel identified in the epithelial cells isolated from the endolymphatic sac of guinea pigs.

We identified a Ca2+-sensitive cation channel in acutely dissociated epithelial cells from the endolymphatic sac (ES) of guinea pigs using the patch-clamp technique. Single-channel recordings showed that the cation channel had a conductance of 24.0 +/- 1.3 pS (n = 8) in our standard solution. The relative ionic permeability of the channel was in the order K+ = Na+ > Ca2+ >> Cl-. This channel was weakly voltage-dependent but was strongly activated by Ca2+ on the cytosolic side at a concentration of around 1 mm in inside-out excised patches. With cell-attached patches, however, the channel was activated by much lower Ca2+ concentrations. Treatment of the cells, under cell-attached configuration, with ionomycin (10 microm), carbonyl cyanide 3-chlorophenylhydrazone (CCCP, 20 microm), or ATP (1 mm), which increased intracellular Ca2+ concentration ([Ca2+]i), activated the channel at an estimated [Ca2+]i from 0.6 microm to 10 microm. It is suggested that some activators of the channel were deteriorated or washed out during the formation of excised patches. Based on this Ca2+ sensitivity, we speculated that the channel contributes to the regulation of ionic balance and volume of the ES by absorbing Na+ under certain pathological conditions that will increase [Ca2+]i. This is the first report of single-channel recordings in endolymphatic sac epithelial cells.

Modulation of intestinal permeability by nitric oxide donors: implications in intestinal delivery of poorly absorbable drugs.

The effects of nitric oxide (NO) donors NOC5 [3-(2-hydroxy-1-(methylethyl)-2-nitrosohydrazino)-1-propanamine ] and NOC12 [N-ethyl-2-(1-ethyl-hydroxy-2-nitrosohydrazino)-ethanamine] on the permeability of 5(6)-carboxyfluorescein (CF) across the intestinal membrane were examined by an in vitro Ussing chamber method. The NO donors significantly increased the intestinal permeability of CF and their absorption-enhancing effects were concentration-dependent over the range of 0.01 to 0.1 mM. Regional differences in the absorption-enhancing effects of the NO donors were observed (colon > jejunum). The absorption-enhancing effect of NOC12 reduced as the molecular weights of compounds increased. Therefore, the degree of absorption-enhancing effect of NOC12 was dependent on the molecular weights of compounds. In the pretreatment studies with NOC12 and lactate dehydrogenase release studies, the absorption-enhancing effect of 0.1 mM NOC12 was reversible and less toxic to the colonic membrane. On the other hand, the absorption-enhancing effect of NOC12 was inhibited by the coadministration of 2-(4-carboxyphenyl) 4,4,5, 5-tetramethylimidazole-1-oxyl 3-oxide sodium salt, an NO scavenger, suggesting that NO can regulate the permeability of water-soluble drugs in the gut. Furthermore, NOC12 (0.1 and 1 mM) significantly decreased the transepithelial electrical resistance value of the colonic membrane, suggesting that the absorption-enhancing mechanism of NOC12 may be partly related to the dilation of the tight junction in the epithelium via a paracellular route. These findings suggest that NO donors may be useful to enhance the intestinal absorption of poorly absorbable drugs.

Crescentic glomerulonephritis with antimyeloperoxidase antibodies developing during the course of IgA nephropathy in a patient with rheumatoid arthritis.

Abstract Glomerulonephritis, such as membranous nephropathy, IgA nephropathy, and p-antineutrophil cytoplasmic autoantibody (ANCA)-related crescentic glomerulonephritis, has been shown to occur in rheumatoid arthritis (RA). However, the occurrence of two types of glomerulonephritis in a patient with RA is rarely observed. Here, we describe a patient with RA who developed crescentic glomerulonephritis with antimyeloperoxidase (MPO) antibodies during the course of IgA nephropathy. This case indicates that crescentic glomerulonephritis and IgA nephropathy may occur together in association with p-ANCA in RA.

Rapid detection of Staphylococcus aureus using bioluminescent enzyme immunoassay.

A bioluminescent enzyme immunoassay (BLEIA) method for detecting protein A-bearing Staphylococcus aureus was developed using biotinylated firefly luciferase. The BLEIA was able to detect protein A at one pg ml-1 and 103 cfu ml-1 level of Staph. aureus. The BLEIA showed significant signals with overnight cultures of all 24 Staph. aureus strains, and the BLEIA did not show any significant signals with overnight cultures of all 44 strains of coagulase-negative staphylococci and the other genus bacteria. After 5 h cultivation beginning at approximately 50 cfu ml-1, the BLEIA was able to detect all 35 Staph. aureus strains isolated from healthy humans.