Drp-1, a potential therapeutic target for brain ischaemic stroke.

BACKGROUND AND PURPOSE: The resistance of CA3 neurons to ischaemia and the ischaemic tolerance conferred by ischaemic preconditioning (IPC) are two well-established endogenous neuroprotective mechanisms. Elucidating the molecules involved may help us find new therapeutic targets. Thus, we determined whether dynamin-related protein 1 (Drp-1) is involved in these processes. EXPERIMENTAL APPROACH: In vivo, we subjected rats to either 10 min severe global ischaemia using a four-vessel occlusion (4-VO) model or 2 min IPC before the onset of 4-VO. In vitro, we performed oxygen glucose deprivation (OGD) studies in rat hippocampal neurons. Drp-1 was silenced or inhibited by siRNA or pharmacological inhibitor Mdivi1. To assess whether mitochondrial Drp-1 alters neuronal vulnerability to ischaemic injury, various approaches were used including western blot, immunohistochemistry, immunofluorescence staining and electron microscopy. Hippocampal function was assessed using an open-field test. KEY RESULTS: Mitochondrial dynamin-related protein 1 (mtDrp-1) was selectively induced by ischaemia in hippocampal CA3 neurons. In hippocampal CA1 neurons, mtDrp-1 was not affected by ischaemia but significantly up-regulated by IPC. Suppression of Drp-1 increased the vulnerability of cells to OGD and global ischaemia. Inhibition of Drp-1 in vivo resulted in loss of acquisition and encoding of spatial information, and also prevented ischaemia-induced mitophagy in CA3. Thus mitochondrial-mediated injury was amplified and resistance to ischaemic injury lost. CONCLUSIONS AND IMPLICATIONS: Our findings that Drp-1 increases the resistance of neurons of hippocampal CA3 affected by global ischaemia and contributes to the tolerance conferred by IPC highlight Drp-1 as a potential therapeutic target for brain ischaemic stroke.

Modified maxillomandibular advancement for obstructive sleep apnoea: towards a better outcome for Asians.

Excluding tracheostomy, maxillomandibular advancement (MMA) is the most effective surgical treatment for obstructive sleep apnoea (OSA). However, the anticipated facial changes may prevent acceptance of this procedure by patients with bimaxillary protrusion, a common feature of Asian faces. We therefore developed a modified MMA technique for such cases, consisting of anterior segmental osteotomies together with standard Le Fort I and bilateral sagittal split osteotomies. A prospective study of 20 consecutive Taiwanese adults with moderate-to-severe OSA who underwent modified MMA and postsurgical orthodontics was undertaken to evaluate the efficacy with regard to OSA and the postoperative facial appearance and dental occlusion. After modified MMA, the mean apnoea-hypopnoea index decreased from 41.6±19.2 n/h to 5.3±4.0 n/h (P<0.001). All patients had a successful outcome. No patient was dissatisfied with their postoperative facial appearance. The mean Peer Assessment Rating score decreased from 21.9±14.3 to 1.7±1.6 (P=0.001). The data suggest that the modified MMA is effective in treating patients with moderate-to-severe OSA without negatively affecting facial appearance or dental occlusion. To achieve a better outcome, surgical-orthodontic integration is warranted. The surgery-first approach can achieve early improvement.

Changes of snoring sound after relocation pharyngoplasty for obstructive sleep apnoea: the surgery reduces mean intensity in snoring which correlates well with apnoea-hypopnoea index.

OBJECTIVE: To investigate objective changes of snoring after surgery in patients with obstructive sleep apnoea (OSA) and correlate these with changes in the apnoea-hypopnoea index (AHI). DESIGN: Prospective case series. SETTING: A novel measurement, Snore Map, was used to analyse full-night snore sounds in terms of the maximal/mean intensity, peak/mean frequency, snoring index and energy type (Snore Map type, 0-4). Snore sound was classified into three bands according to frequency energy spectrum: B1 (40-300 Hz), B2 (301-850 Hz) and B3 (851-2000 Hz). PARTICIPANTS: Thirty-four male and two female OSA patients (mean age, 39 years; mean AHI, 53.1/h; mean body mass index, 26.8 kg/m(2) ) with favourable anatomic structure were consecutively enrolled. MAIN OUTCOME MEASURES: Parameters of polysomnographies and Snore Maps at baseline and six months after operation were compared. Statistical significance was set at P < 0.05. RESULTS: Thirty-two patients completed this study. The mean reduction in the total-snoring index was insignificant but there were significant decreases in total mean intensity, total peak frequency, total mean frequency and Snore Map type after surgery. There were also significant decreases in the mean intensity in all three bands, the snoring index in B2/B3 and the mean frequency in B1 postoperatively. Changes in the total mean intensity, total mean frequency, B2 mean intensity and B3 snoring index positively correlated with change in the AHI. CONCLUSIONS: Relocation pharyngoplasty significantly decreases both the snoring sound intensity and snoring frequency. These reductions are directly proportional to the improvement of OSA.

Do glial cells play an anti-oxidative role in Huntington's disease?

Oxidative stress is a condition of imbalance between reactive oxygen species (ROS) formation and antioxidant capacity as a result of dysfunction of the antioxidant system. ROS can be served as a second messenger at low or moderate concentration, while excessive amount of ROS under oxidative stress condition would destroy macromolecules like proteins, DNA, and lipids, finally leading to cell apoptosis or necrosis. Changes in these macromolecules are involved in various pathological changes and progression of diseases, especially neurodegenerative diseases. Neurodegenerative diseases are morphologically featured by progressive neuronal cell loss, accompanied with inclusions formed by protein aggregates in neurons or glial cells. Neurons have always received much more attention than glial cells in neurodegenerative diseases. Actually, glial cells might play a key role in the functioning of neurons and cellular survival through an antioxidant way. Additionally, neurons can modulate the activities of glia either. Herein, the main purposes of this review are to mention the connection between Huntington's disease (HD) and oxidative stress, to summarize the characteristics and functions of glial cells in HD, to state the cross talk between neurons and glial cells, and to emphasize the conclusive role of activation of Keap1-Nrf2-ARE pathway in glial cells against oxidative stress in HD.

Glucosamine induces activated T cell apoptosis through reduced T cell receptor.

Glucosamine (GlcN), like N-acetylglucosamine (GlcNAc), is salvaged into the hexosamine pathway and is converted to UDP-GlcNAc. Golgi N-glycan branching enzymes produce N-glycans, using UDP-GlcNAc as a substrate, which attach to the T cell receptor (TCR) and cytotoxic T-lymphocyte antigen-4 (CTLA-4). These findings suggest that GlcN exerts the immunoregulation through TCR signalling, which could be involved not only in cytokine production but also activated T cell apoptosis. In fact, a preliminary study showed that GlcN reduced the number of CD3+ T cells of NC/Nga mice with AD-like skin lesions. Therefore, whether apoptosis of T cells would be one of the potential molecular mechanisms of GlcN-induced immunosuppression was investigated. Cultured human primary along with Jurkat T cells and purified T cells from NC/Nga mice with or without Df-induced AD-like skin lesion were used for the study. Glucosamine treatment increased the number of T cells expressing ß1,6GlcNAc-branched N-glycans, with reduced ZAP-70 phosphorylation and enhanced CTLA-4 expression. Glucosamine treatment reduced the number of activated T cells from both the human primary and Jurkat cells and the dermatitis-induced mice. The expression of FasL and activated caspases, particularly caspase-3, was increased, whereas the phosphorylation of PI3K, Akt and NF-κB was decreased by GlcN treatment. Therefore, in addition to down-regulating TCR signalling and promoting CTLA-4 expression, GlcN may also suppress T cell function by enhancing apoptosis of activated T cells, through both extrinsic and intrinsic apoptotic signalling pathways, which were regulated by the inhibition of PI3K/Akt and NF-κB phosphorylation.

Claulansine F suppresses apoptosis induced by sodium nitroprusside in PC12 cells.

Reactive oxygen species (ROS) are known to be involved in many neurodegenerative diseases. This study assessed the effect of Claulansine F, a new carbazole isolated from Clausena lansium, on sodium nitroprusside (SNP)-treated rat pheochromocytoma PC12 cells. First, it was found that Claulansine F showed more potential on inhibiting the programmed death of PC12 cells than edaravone by cell viability, morphologic observation, and flow cytometric analysis. Further results also showed that Claulansine F attenuated the production of total intracellular ROS formation and lipid peroxidation in PC12 cells, inhibited the mitochondrial membrane potential (MMP) loss, and prevented the programmed cell death event via the P53/Bcl-2 family pathway. Its protective effect was likely medicated by the hydroxyl radical (·OH) scavenging ability, as it appeared to be not involved in the natural antioxidant system. These results suggested a promising potential for Claulansine F as a ROS scavenger in pathologies, where an oxidative stress is involved.

IMM-H004, a novel coumarin derivative compound, protects against amyloid beta-induced neurotoxicity through a mitochondrial-dependent pathway.

We have investigated the effect of IMM-H004 (7-hydroxy-5-methoxy-4-methyl-3-(4-methylpiperazin-1-yl)-2H-chromen-2-one), a coumarin derivative, on the amyloid beta (Aß)-induced neurotoxicity in primary culture cortical neurons and pheochromocytoma (PC12) cells. Our results showed that treatment with IMM-H004 markedly reduced the number of apoptotic cells after exposure to Aß25-35 or Aß1-42, determined by MTT, TUNEL staining and Flow cytometry. Further study indicated that IMM-H004 significantly inhibited Aß-induced cytotoxicity and apoptosis by reversing Aß-induced mitochondrial dysfunction, including MMP (mitochondrial membrane potential) decrease, reactive oxygen species production, and mitochondrial release of cytochrome c. IMM-H004 can regulate the interaction between Bax and Bcl-2, decreased levels of p53 and active caspase-3 protein induced by Aß25-35. Furthermore, IMM-H004 also reduced translocation of AIF (apoptosis-inducing factor) induced by Aß25-35. These results demonstrated that IMM-H004 was capable of protecting neuronal cells from Aß-induced degeneration through a mitochondrial-dependent apoptotic pathway. The results of this study lend further credence to the notion that IMM-H004 is a 'multipotent therapeutic agrent' that reduces toxic levels of brain Aß, and holds the potential to protect neuronal mitochondrial function in Alzheimer's disease.

Occurrence and distribution of Naegleria species from thermal spring environments in Taiwan.

UNLABELLED: Naegleria spp. is a free-living amoeba that can be found in the natural environment. A number of Naegleria spp. can cause fatal infections in the central nervous system in humans and animals, and the most important source of infection is through direct water contact. In this study, water samples from various thermal springs were taken from four thermal spring areas. Naegleria spp. was detected via culture confirmation and molecular taxonomic identification. Among the 60 samples obtained, Naegleria spp. was identified in 26 (43·3%) samples. The identified species included Naegleria australiensis, Naegleria gruberi, Naegleria lovaniensis and Naegleria mexicana. The presence of living Naegleria spp. was significantly associated with elevated pH value in the water sample. SIGNIFICANCE AND IMPACT OF STUDY: In this study, we examined the presence of living Naegleria spp. in thermal spring waters in south-eastern Taiwan. Naegleria spp. was isolated and culture-confirmed from thermal spring water. Naegleria fowleri was not found in all water samples, and Naegleria australiensis was the most common Naegleria genotype.

Protopanaxatriol metabolites identified by LC-MS/MS after oral administration in mice.

OBJECTIVE: 20(S)-Protopanaxatriol (Ppt), a well-known end metabolite of protopanaxatriol-type saponins, has recently been reported to have the same bioactivity as its prototype. Whether or not Ppt could be further metabolized into other compounds in vivo is still unknown. The present study is aimed to determine the structures of Ppt metabolites in mice. MATERIALS: The metabolites were produced by intragastric gavage of Ppt in mice. The homogenate of small intestine was used for analysis after solid phase extraction. METHODS: The metabolic profile of Ppt was investigated by using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), which were also used to identify the structures of metabolites. Accurate mass measurement using LC-time of flight MS was applied to determine the element composition of metabolites and thus to confirm their proposed structures. RESULTS: One Phase I and three Phase II metabolites were detected at 1 h, 5 h, and 10 h after administration of Ppt, which were the same at the three time points. The Phase I metabolic changes observed included dehydrogenation and hydroxylation of the steroid-like structure, as well as formation of an ester bond at C-20 of the side chain. The Phase II metabolites involved conjugation to aminoethylsulfonic acid after hydrolysis of the ester bond. A possible biotransformation pathway was proposed. CONCLUSIONS: Ppt yielded four metabolites in vivo, and 1 h was enough to complete the biotransformation process of Ppt.

Effects of anti-bursin monoclonal antibody on immunosuppression in the duck (Cherry Valley duck).

To study the immunologic function of bursin, we analyzed the effects of anti-bursin monoclonal antibody (mAb) on the immunosuppression in ducks (Cherry Valley duck) by injecting various doses of the anti-bursin mAb into 13-d duck embryos. After hatch, cell-mediated immune activity and humoral responses were studied using lymphocyte proliferation test, tube agglutination test, and indirect enzyme-linked immunosorbent assay to detect anti-Escherichia coli antibodies and antibodies to Riemerella anatipestifer, respectively. Simultaneously, relative weights (BW-adjusted) of bursa of Fabricius (BF), spleen, and thymus were determined. Additionally, the morphology of BF, spleen, and thymus was examined at various ages using conventional histology. Follicle morphology of BF was analyzed by image analysis. The results indicated that anti-bursin mAb markedly decreased duck lymphocyte proliferation, the antibody-producing ability to bacteria, as well as the relative BF weight. Moreover, the anti-bursin mAb hindered the development of BF follicles.

Combined bronchoalveolar lavage and polymerase chain reaction in the diagnosis of pulmonary tuberculosis in smear-negative patients.

SETTING: The polymerase chain reaction (PCR) may be sensitive and specific for the diagnosis of tuberculosis, but most reports are of studies conducted in well-controlled laboratories. A study to evaluate the clinical value of bronchoalveolar lavage (BAL) combined with PCR was necessary. OBJECTIVE: One hundred and thirty one patients were recruited into the study from March 1994 to February 1997. DESIGN: Patients with a positive acid-fast stain on sputum smear were recruited into group A as positive controls, patients with lung cancer and a negative acid-fast stain on sputum smear were put into group B as negative controls, and patients who had clinical symptoms of pulmonary TB without sputum or with negative smear results were the investigating group. PCR was performed on the sputum samples from group A and B patients and on the BAL fluid from those in group C. RESULTS: The sensitivity of PCR was 96% in group A, and the specificity was 100% in group B. The sensitivity of PCR in the BAL fluid from the group C patients was 36% and the specificity was 96%; the positive predictive value was 94% and the negative predictive value was 45%. CONCLUSION: BAL plus PCR is useful in the rapid diagnosis of pulmonary TB in non-productive or smear-negative patients.

Neuronal activity in dorsomedial frontal cortex and prefrontal cortex reflecting irrelevant stimulus dimensions.

Previous studies of the dorsomedial frontal cortex (DMF) and the prefrontal cortex (PF) have shown that, when monkeys respond to nonspatial features of a discriminative stimulus (e.g., color) and the stimulus appears at a place unrelated to the movement target, neurons nevertheless encode stimulus location. This observation could support the idea that these neurons always encode stimulus location, regardless of its relevance to an instrumentally conditioned behavior. Past studies, however, leave open the possibility that activity observed during one operant task might reflect the contingencies of a different task, performed at different times. To test these alternatives, we examined the activity of DMF and PF neurons in two rhesus monkeys conditioned to perform an operant eye-movement task in which only the color and shape of visual stimuli served as salient discriminative features. Each of eight stimuli was associated with a response to a different eye-movement target. The location of these stimuli varied from trial to trial but was of no behavioral relevance, and the monkeys did not perform any operant task in which stimulus location controlled behavior. A substantial minority of neurons in both DMF and PF nevertheless encoded stimulus location, which indicates that this property does not depend on its relevance in an instrumentally conditioned behavior.

Spontaneous variability of cardiac output in ventilated critically ill patients.

OBJECTIVE: To define the magnitude of spontaneous cardiac output variability over time in sedated medical intensive care unit patients attached to a continuous cardiac output monitor, and to determine whether high level positive end-expiratory pressure or inverse inspiratory-to-expiratory (I:E) ratio ventilation resulted in greater variability over time than low positive end-expiratory pressure with conventional I:E ratio ventilation. DESIGN: Prospective study. SETTING: Medical intensive care unit in a tertiary medical center. PATIENTS: A total of 22 hemodynamically stable acute respiratory failure patients with a pulmonary artery catheter inserted for hemodynamic monitoring INTERVENTIONS: After being sedated, patients were randomized ultimately to receive pressure control ventilation first at setting A (high positive end-expiratory pressure [15 cm H2O] with conventional I:E ratio [1:2]) and then at setting B (low positive end-expiratory pressure [5 cm H2O] with inverse I:E ratio [2:1]), or vice versa, and then at setting C (low positive end-expiratory pressure [5 cm H2O] with conventional I:E ratio [1:2]). Each ventilation setting period lasted 1 hr. MEASUREMENTS AND MAIN RESULTS: Cardiac output (CO) was measured continuously. The continuous CO value displayed was updated every 30-60 secs. The updated value reflected an average of the previous 3-6 mins. The coefficient of variation (CV) of CO for each setting in each patient was calculated to represent the spontaneous variability. The mean CO+/-SD and CV of each setting was 5.7+/-1.8 L/min and 4.4% for setting A, 5.6+/-1.5 L/min and 4.6% for setting B, and 5.9+/-1.7 L/min and 4.8% for setting C. Analysis of variance revealed no significant differences between the CVs of the three settings. The 95% confidence interval for the COs for each setting was approximately the mean CO+/-0.1 x mean CO measured. CONCLUSIONS: In critically ill sedated medical intensive care unit patients with stable hemodynamics, the spontaneous variability of cardiac output over time was not significant. High positive end-expiratory pressure (15 cm H2O) and inverse ratio ventilation (2:1) did not contribute to increased spontaneous variability of cardiac output.

Tuberculin purified protein derivative up-regulates the telomerase activity of peripheral blood mononuclear cells from patients with pulmonary tuberculosis.

Telomerase activity was detectable in cells of tuberculous pleural effusions at high percentage. To investigate the possible role of telomerase in the immune function, we examined the proliferating state and the expression of telomerase activity in peripheral blood mononuclear cells (PBMC) from 13 patients with pulmonary tuberculosis (TB) and 13 healthy volunteers in response to tuberculin purified protein derivative (PPD) challenge. Exposure of cells to phytohemagglutinin (PHA) significantly promoted PBMC proliferation during a 6 day-period in both TB patient and healthy volunteer groups. PPD treatment also significantly promoted PBMC proliferation during a 6 day-period in TB patient group, but had no significant effect in healthy volunteer group. During the same period, telomerase activity was detected in every PHA- and PPD-treated samples of the TB patient group. However, the telomerase activity was not detected in PPD-treated samples from healthy donors and all the untreated samples. Our results indicate that the telomerase activity in PBMC could be induced by PPD stimulation in TB patients. Telomerase activity may thus play a permissive role in cell division and clonal expansion of the immune cells in response to TB.

Spontaneous variability of arterial oxygenation in critically ill mechanically ventilated patients.

OBJECTIVE: To assess the magnitude of spontaneous variability of arterial oxygenation and oxygen tension-based indices over time in medical intensive care unit (ICU) patients and to study whether high positive end-expiratory pressure (PEEP) or inverse inspiratory-to-expiratory (I:E) ratio ventilation (IRV) results in a greater variability than low PEEP with conventional I:E ratio ventilation. DESIGN: Prospective study. SETTING: Medical ICU in a tertiary medical center. PARTICIPANTS: 23 patients requiring a pulmonary artery floating catheter for hemodynamic monitoring. INTERVENTION: After being completely sedated, patients were randomized to receive pressure-control ventilation at setting A: high PEEP (15 cm H2O) with conventional I:E ratio (1:2) and setting B: inverse I:E ratio (2:1) with low PEEP (5 cm H2O) alternately, and then at setting C: low PEEP (5 cm H2O) with conventional I:E ratio (1:2). Each ventilation setting lasted 1 h. MEASUREMENTS AND RESULTS: The arterial and mixed venous blood samples were measured simultaneously at baseline (time 0), and at 15, 30, 45, and 60 min thereafter. The coefficient of variation (CV) of arterial oxygen tension (PaO2) over time was 5.9 % for setting A, 7.2 % for setting B, and 6.9 % for setting C. ANOVA showed no significant differences in CVs of PaO2 between the three settings. Oxygen tension-based indices, alveolar-arterial oxygen difference (A-aDO2) and PaO2/ PAO2 (alveolar oxygen tension), displayed CVs equal to that of PaO2; the CV of A-aDO2/PaO2 was significantly greater than that of PaO2. CONCLUSIONS: In critically ill medical ICU patients, despite sedation, the spontaneous variability in PaO2 over time is substantial. A high PEEP or IRV does not contribute to the increased variation in PaO2.

Serodiagnosis of tuberculosis by enzyme-linked immunosorbent assay for anti-A60 and anti-A38.

BACKGROUND: For early diagnosis of tuberculosis (TB), especially in the patients without adequate sputum specimens for examination, we found a simple, rapid and inexpensive method among many current available diagnostic tools, the enzyme-linked immunosorbent assay (ELISA). To investigate the diagnostic effectiveness of this method, we applied ELISA for detection of antigen 60 IgG and IgM as well as antigen 38 IgG antibodies at Chang Gung Memorial Hospital from April 1995 through June 1996. MATERIALS AND METHODS: Sixty-seven patients were enrolled and divided into 3 groups, Group A (n = 24), patients with positive sputum acid-fast stain; Group B (n = 18), patients with lung cancer and negative sputum acid-fast stain; and Group C (n = 25), patients with chest roentgenogram (CXR) which were suggestive of TB but with negative acid-fast stain results or no sputum for examination. RESULTS: For the A60 IgG antibody, we found a sensitivity rate of 91.7% for Group A and Group B, and 85.7% for Group C as well as an overall sensitivity of 89.5% but with lower specificity. For the A60 IgM antibody, a lower sensitivity (37.5%, 14.3%, 28.9%, respectively) was found but with higher specificity. For the A38 IgG antibody, we found a lower sensitivity (40%, 11.1%, 31%, respectively) but with higher specificity (100%, 71.4%, 90%, respectively). CONCLUSION: With a high sensitivity but low specificity for diagnosis of TB, A60 IgG ELISA could be used as a rapid, simple screening test for patients with results suggestive of TB, especially in those who had no sputum or had negative sputum acid-fast stain results. Otherwise, A60 IgM or A38 IgG ELISA, with a high specificity, could be used as a reliable test in the diagnosis of pulmonary TB when the result is positive. In summary, although ELISA is a simple, rapid, inexpensive method, it is helpful but limited in the diagnosis of pulmonary TB.