A protonic biotransducer controlling mitochondrial ATP synthesis.

In nature, protons (H+) play an important role in biological activities such as in mitochondrial ATP synthesis, which is driven by a H+ gradient across the inner membrane, or in the activation of acid sensing ion channels in neuron cells. Bioprotonic devices directly interface with the H+ concentration (pH) to facilitate engineered interactions with these biochemical processes. Here we develop a H+ biotransducer that changes the pH in a mitochondrial matrix by controlling the flow of H+ between a conductive polymer of sulfonated polyaniline and solution. We have successfully modulated the rate of ATP synthesis in mitochondria by altering the solution pH. Our H+ biotransducer provides a new way to monitor and modulate pH dependent biological functions at the interface between the electronic devices and biological materials.

Synthesis of a fluorescent photoaffinity probe of OSW-1 by site-selective acylation of an inactive congener and biological evaluation.

A novel fluorescent photoaffinity probe of OSW-1 was prepared in two steps from a naturally occurring inactive congener by a sequential site-selective acylation strategy using Me2SnCl2. It displayed highly potent anticancer activity and a similar intracellular localization property to that of a fluorescently-tagged OSW-1, thereby demonstrating its potential utility in live cell studies.

'PrP systemic deposition disease': clinical and pathological characteristics of novel familial prion disease with 2-bp deletion in codon 178.

BACKGROUND AND PURPOSE: A novel TYPE of prion disease associated mainly with autonomic-sensory polyneuropathy was reported by us previously. METHODS: Here the autopsy pathology for patient 1 (the sister) and the clinical characteristics of her younger brother (patient 2) are newly reported. Polymerase chain reaction based restriction fragment length polymorphism analysis of the prion protein gene (PRNP) was performed on both patients and their father (normal control). RESULTS: Polymerase chain reaction based restriction fragment length polymorphism analysis revealed a 2-bp deletion (CT) in codon 178 that causes an additional variable 25 amino acids at the C terminal, from the mutation site to the premature stop codon at codon 203, in both patients 1 and 2 but not in their father. The autopsy of patient 1 showed remarkable prion protein (PrP) deposits in the sympathetic ganglion and peripheral nerves, correlated to her severe autonomic sensory failure. PrP deposits were also found in the central nervous system and peripheral organs such as the heart, lung, stomach, jejunum, ileum, colon, urinary bladder and adrenal gland. The symptoms and biopsy findings of patient 2 were nearly the same as those reported previously for patient 1. His cognitive function was well preserved, but autonomic functions were severely impaired. His biopsied samples showed PrP deposits in the sural nerve and nerve plexuses of the stomach and colon. CONCLUSION: The present unique 2-bp deletion (CT) in codon 178 induced a 'PrP systemic deposition disease' such as pan-autonomic failure, sensory neuropathy and mild cognitive impairment with a specific pathology.

Feeding hydroalcoholic extract powder of Lepidium meyenii (maca) increases serum testosterone concentration and enhances steroidogenic ability of Leydig cells in male rats.

Although Lepidium meyenii (maca), a plant growing in Peru's central Andes, has been traditionally used for enhancing fertility and reproductive performance in domestic animals and human beings, effects of maca on reproductive organs are still unclear. This study examined whether feeding the hydroalcoholic extract powder of maca for 6 weeks affects weight of the reproductive organs, serum concentrations of testosterone and luteinising hormone (LH), number and cytoplasmic area of immunohistochemically stained Leydig cells, and steroidogenesis of cultured Leydig cells in 8-week-old male rats. Feeding the extract powder increased weight of seminal vesicles, serum testosterone level and cytoplasmic area of Leydig cells when compared with controls. Weight of prostate gland, serum LH concentration and number of Leydig cells were not affected by the maca treatment. The testosterone production by Leydig cells significantly increased when cultured with 22R-hydroxycholesterol or pregnenolone and tended to increase when cultured with hCG by feeding the extract powder. The results show that feeding the hydroalcoholic extract powder of maca for 6 weeks increases serum testosterone concentration associated with seminal vesicle stimulation in male rats, and this increase in testosterone level may be related to the enhanced ability of testosterone production by Leydig cells especially in the metabolic process following cholesterol.

Frequency of HER2 expression of circulating tumour cells in patients with metastatic or recurrent gastrointestinal cancer.

BACKGROUND: The clinical significance of circulating tumour cell (CTC) detection in gastrointestinal (GI) cancer remains controversial and the molecular biological characteristics of CTCs are poorly understood. METHODS: A total of 87 patients with metastatic or recurrent GI cancer were prospectively enrolled. Circulating tumour cells and their HER2 status were assessed using the CellSearch System. RESULTS: Among the 62 CTC-positive cases, we found 22 discordant cases (35.5%). Among the HER2-negative primary tumours, 17 of 54 developed HER2-positive CTCs. Five of eight had HER2-negative CTCs among the HER2-positive primary tumours. CONCLUSION: The findings in the current study suggest that it is critical to evaluate the HER2 status of not only the primary tumour but also the CTCs because the metastasising tumour cells are the primary target of systemic therapy.

Non-targeted analyses of animal plasma: betaine and choline represent the nutritional and metabolic status.

Simple liquid chromatography-mass spectrometry (LC-MS) was applied to non-targeted metabolic analyses to discover new metabolic markers in animal plasma. Principle component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA) were used to analyse LC-MS multivariate data. PCA clearly generated two separate clusters for artificially induced diabetic mice and healthy control mice. PLS-DA of time-course changes in plasma metabolites of chicks after feeding generated three clusters (pre- and immediately after feeding, 0.5-3 h after feeding and 4 h after feeding). Two separate clusters were also generated for plasma metabolites of pregnant Angus heifers with differing live-weight change profiles (gaining or losing). The accompanying PLS-DA loading plot detailed the metabolites that contribute the most to the cluster separation. In each case, the same highly hydrophilic metabolite was strongly correlated to the group separation. The metabolite was identified as betaine by LC-MS/MS. This result indicates that betaine and its metabolic precursor, choline, may be useful biomarkers to evaluate the nutritional and metabolic status of animals.

Maxillary haemangioma successfully resected by endoscopic approach.

OBJECTIVE: We report an extremely rare case of maxillary haemangioma. METHOD: Case report and review of the literature concerning haemangioma arising from the nasal cavity and paranasal sinuses. RESULTS: Maxillary haemangioma is rare and sometimes requires wider resection than nasal haemangioma if a large tumour is found. We present a case of maxillary haemangioma in a 37-year-old Japanese woman, which was completely resected by pre-operative embolisation and endoscopic sinus surgery. CONCLUSION: Our findings suggest that if a large maxillary haemangioma is diagnosed pre-operatively, the treatment of choice is pre-operative embolisation followed by endoscopic sinus surgery, in order to avoid the surgical complications associated with wide resection.

Developmental effects: oestrogen-induced vaginal changes and organotin-induced adipogenesis.

The emerging paradigm, the foetal origin of adult disease, is a new framework for considering the effects of endocrine disrupters on human and animal health. Prenatal diethylstilbestrol (DES) exposure resulted in various reproductive tract abnormalities in women, which is called as DES syndrome. Similar abnormalities have been demonstrated in experimental animals exposed perinatally to oestrogens. Developmental oestrogen exposure induces persistent proliferation of vaginal epithelial cells in mice. The persistent changes in the vagina of mice neonatally exposed to oestrogens results from persistent phosphorylation of erbB2 and oestrogen receptor alpha, sustained expression of EGF-like growth factors and phosphorylation of JNK1, IGF-I receptor and Akt. The ubiquitous environmental contaminant, tributyltin chloride (TBT) is well known to induce the development of male sex characteristics (imposex) in gastropods. We recently found that TBT and its congeners induce the differentiation of adipocytes in vitro and increase adipose mass in vivo in vertebrates. TBT is a nanomolar affinity ligand for retinoid X receptor (RXR) in the rock shell and for both the RXRalpha and the peroxisome proliferator-activated receptor gamma (PPARgamma) in the amphibian (Xenopus laevis), mouse, and human. TBT promotes adipogenesis in the murine 3T3-L1 cell model and perturbs key regulators of adipogenesis and lipogenic pathways in vivo, primarily through activation of RXRalpha and PPARgamma. Moreover, in utero exposure to TBT leads to strikingly elevated lipid accumulation in adipose depots, liver, and testis of neonate mice and results in increased adipose mass in adults. In X. laevis, ectopic adipocytes form in and around gonadal tissues following organotin, RXRalpha or PPARgamma ligand exposure. TBT represents the first example of an environmental endocrine disrupter that promotes adverse effects from gastropods to mammals. Prenatal (TBT) and early postnatal exposures (oestrogens) stand as strong examples of endocrine disrupting compounds that permanently alter developmental programming.

Predicting recurrence following curative surgery in stage I non-small cell lung cancer patients using an angiogenesis-associated factor.

There are still difficulties in determining the risk of recurrence to decide whether to perform selective adjuvant treatment for stage I non-small cell lung cancer. This study reviewed 122 stage I patients who underwent curative surgery to assess the usefulness of an angiogenesis-associated factor as a predictor of recurrence. By immunohistochemical examination, we collected information about tumor-induced vascular endothelial growth factor (VEGF)-A and -C expression at the primary site. During the median follow-up period of 120 months, the overall 10-year survival rate was 53.5%. The 10-year survival rates according to VEGF-A/C expression were as follows: VEGF-A high/VEGF-C high group, 26.0%; VEGF-A high/VEGF-C low group, 42.7%; VEGF-A low/VEGF-C high group, 73.1%; VEGF-A low/VEGF-C low group, 65.1%. The VEGF-A high/VEGF-C high group showed the worst outcome. The diagnostic values of the marker combination for predicting recurrence were as follows: sensitivity, 71.4%; specificity, 63.8%; and accuracy, 65.6%. When stratified by T factor, preferable high values for both negative predictive value and specificity were obtained in patients with stage IA disease. In order to select the patients eligible for selective adjuvant therapy at early stages, especially in stage IA disease, simultaneous assessment of tumor-induced VEGF-A/C warrants further study.

In vivo experimental model of human gingival mucosa using immunodeficient mice.

BACKGROUND AND OBJECTIVE: To establish an in vivo experimental model for examining human periodontal tissue, the present study examined several transplant techniques that maintain the structure and characteristics of human gingival mucosa. MATERIAL AND METHODS: Human oral mucosal tissue samples were collected from the gingiva (n = 11), palate (n = 1), and tongue (n = 3). These mucosal grafts were transplanted onto BALB/c nu/scid mice with double-mutant immunodeficiency. Murine skin, twice the size of the graft, was cut open in an ' square superset'-shape. Next, the connective tissue side of the graft was placed onto the murine connective tissue. Immunohistochemical analysis was also performed, using polyclonal rabbit antibody to involucrin, monoclonal antibody to vimentin, monoclonal antibody to CD34, and monoclonal antibody to Ki-67, to determine whether the characteristics of human oral mucosa were maintained. RESULTS: When the connective tissue side of the graft was placed on the murine fascial membrane, the histological structure of the graft was maintained for 60 d. These grafts were examined for human characteristics using human-specific antibodies. Immunohistochemically, the expression patterns of involucrin, vimentin, and Ki-67 indicated that transplanted mucosa revealed normal human characteristics, including differentiation and proliferation up to 80 d. CD34 was not detected in the graft endothelial cells. CONCLUSION: The present study revealed that the novel technique of transplantation of human gingival mucosa in nu/scid mice may serve as an in vivo experimental model of periodontal disease.

Long-term treatment with TGFbeta1 impairs mechanotransduction in bovine aortic endothelial cells.

BACKGROUND AND PURPOSE: Vascular endothelial cells play a role in the physiological response to mechanical stress. Transforming growth factor beta1 (TGFbeta1) induces morphological changes in endothelial cells, and this may alter their mechanosensitive responses. The aim of this study was to examine the effects of TGFbeta1 on hypotonic stress (HTS)-induced responses in bovine aortic endothelial cells (BAECs). EXPERIMENTAL APPROACH: Cultured BAECs were treated with 3 ng ml(-1) TGFbeta1 for 24 h (24h-TGFbeta1) or 7 days (7d-TGFbeta1). Cytosolic actin fibres were stained with rhodamine-phalloidin. Intracellular Ca2+ concentration was measured using fura2. Tyrosine phosphorylation and RhoA expression were assessed by Western blotting. Expression of RhoA mRNA was assessed by real-time PCR. KEY RESULTS: BAECs developed pseudopod-like processes within 24 h and showed a fibroblast-like appearance after 7 days. HTS induced Ca2+ transients via endogenous ATP release in both control and 24h-TGFbeta1 BAECs but not in 7d-TGFbeta1 BAECs. We have previously shown that HTS-induced ATP release is mediated by sequential activation of RhoA and tyrosine kinases. The basal amount of membrane-bound RhoA was significantly lower in 7d-TGFbeta1 than in 24h-TGFbeta1 or control BAECs. HTS increased the membrane-bound RhoA to the same fractional level in 24h-TGFbeta1 and control BAECs, but its net maximal amount was significantly lower in 7d-TGFbeta1. HTS-induced downstream signals of RhoA activation, i.e. the tyrosine phosphorylation of FAK and paxillin, were markedly suppressed in 7d-TGFbeta1 BAECs. CONCLUSIONS AND IMPLICATIONS: These results indicate that long-term treatment with TGFbeta1 does not impair mechanoreception in BAECs but impairs mechanotransduction by affecting RhoA membrane translocation.

Thoracoscopic robot-assisted bronchoplasty.

BACKGROUND: In robotic surgery, the ideal position of the system, as well as the optimal working angles and the proper positioning of the thoraco ports position is very important. No robot-assisted bronchoplasty has been reported. Our study describes use of the da VinciTM surgical system (Intuitive Surgical, Inc.) for robotic sleeve upper lobectomy in a human fresh cadaver. METHODS: A male cadaver was placed in the left lateral decubitus position. After thoracoscopic upper lobectomy was performed through the working port and the two ports, the robotic system was then set up behind the cadaver. The working port allowed introduction of the optical scope and the robotic surgical arms were inserted into the thoraco ports. The right bronchus was dissected and wedge was cut out with the robotic scissors. After standard lymph node dissection, end-to-end bronchial anastomosis was performed with robotic instruments. Once the anastomosis was complete, air leakage was checked with saline solution placed in the pleural cavity. RESULTS: Thoracoscopic robot-assisted bronchoplasty was performed successfully. CONCLUSIONS: In evaluating various positions of the system we demonstrated that our technique is sufficient approaches to robotic bronchoplasty. This procedure offers specific advantages over conventional bronchoplasty with accuracy and safety.

Evaluation of carcinomatous pleuritis on initial relapse in patients with lung cancer who underwent complete resection.

The purpose of this study was to evaluate the clinical characteristics and results of lung cancer patients with carcinomatous pleuritis on initial relapse. Over the past decade, seven patients out of 595 surgically treated lung cancer patients developed carcinomatous pleuritis as the initial sign of relapse. All patients had undergone complete resection of the primary peripheral type adenocarcinoma. Only 3 cases showed obvious visceral pleural invasion, while venous or lymphatic invasion were frequently found in the histological sections. The mean disease-free interval after surgery was 30.9 months. The median survival after the development of carcinomatous pleuritis was 14 months. The results showed the possible association of blood/lymphatic routes with the development of this type of recurrence. In some lung cancer patients with carcinomatous pleuritis on initial relapse, long-term survival can be expected by aggressive interventions consisting of intrathoracic and/or systemic chemotherapies.