Utilising TMS-EEG to Assess the Response to Cerebellar-Brain Inhibition.

Cerebellar-brain inhibition (CBI) is a transcranial magnetic stimulation (TMS) paradigm indexing excitability of cerebellar projections to motor cortex (M1). Stimulation involved with CBI is often considered to be uncomfortable, and alternative ways to index connectivity between cerebellum and the cortex would be valuable. We therefore sought to assess the utility of electroencephalography in conjunction with TMS (combined TMS-EEG) to record the response to CBI. A total of 33 volunteers (25.7 ± 4.9 years, 20 females) participated across three experiments. These investigated EEG responses to CBI induced with a figure-of-eight (F8; experiment 1) or double cone (DC; experiment 2) conditioning coil over cerebellum, in addition to multisensory sham stimulation (experiment 3). Both F8 and DC coils suppressed early TMS-evoked EEG potentials (TEPs) produced by TMS to M1 (P < 0.05). Furthermore, the TEP produced by CBI stimulation was related to the motor inhibitory response to CBI recorded in a hand muscle (P < 0.05), but only when using the DC coil. Multisensory sham stimulation failed to modify the M1 TEP. Cerebellar conditioning produced changes in the M1 TEP that were not apparent following sham stimulation, and that were related to the motor inhibitory effects of CBI. Our findings therefore suggest that it is possible to index the response to CBI using TMS-EEG. In addition, while both F8 and DC coils appear to recruit cerebellar projections, the nature of these may be different.

Elevated hydrostatic pressure enhances the motility and enlarges the size of the lung cancer cells through aquaporin upregulation mediated by caveolin-1 and ERK1/2 signaling.

The mechanical characteristics presented in cancer microenvironment are known to have pivotal roles in cancer metastasis, which accounts for the leading cause of death from malignant tumors. However, while a uniformly distributed high interstitial fluid pressure (IFP) is a common feature in solid tumors, the effects of high IFP on the motility and invasiveness of cancer cells remain obscure. Using cell-culture devices that simulated increased IFP conditions by applying hydrostatic pressure (HP) ranging from 0 to 20 mm Hg to the cells, we found that the elevated HPs increased the migration speeds, invasiveness, cell volume, filopodial number and aquaporin-1 (AQP1), Snail and vinculin expression levels, as well as phosphorylation of caveolin-1 and extracellular signal-regulated kinase1/2 (ERK1/2), in the lung cancer cells CL1-5 and A549. The increases of migration speed and cell volume correlated temporally with the increase of AQP1 expression. The elevated HP-induced migration acceleration was hindered by AQP1 knockdown using small interfering RNA (siRNA) transfection. Inhibition of ERK1/2 phosphorylation using the mitogen-activated protein kinase kinase inhibitor PD98059 abrogated the elevated HP-induced AQP1 upregulation and migration acceleration in the cancer cells. Caveolin-1 knockdown by siRNA transfection attenuated the HP-induced, ERK1/2-depedent AQP1 upregulation and migration acceleration. Further biochemical studies revealed that the caveolin-1 activation-driven ERK1/2 signaling is mediated by Akt1/2 phosphorylation. By contrast, the elevated HPs had negligible effects on the migration speed and volume of normal bronchial epithelial cells. These results disclose a novel mechanism relating high IFP to the invasiveness of cancer cells and highlight potential targets to impede cancer spreading.

Active immunization of fatty acid translocase specifically decreased visceral fat deposition in male broilers.

Lipid accumulation of avian adipocytes is mainly dependent upon the fatty acid transmembrane uptake process mediated by membrane proteins, such as fatty acid translocase (FAT/CD36), fatty acid transport protein 1, and caveolin-2. To examine the effects of FAT/CD36 on spatial-specific fat deposition, 60 broiler chickens were randomly allocated to 2 groups by sex. Each male or female group contained 2 subgroups (n = 14-15) inoculated by intramuscular injection with chicken FAT/CD36 or BSA (control) immunogens at 34, 49, and 63 d. The subcutaneous and visceral fat deposits were measured, as were levels of plasma triglyceride and free fatty acid. Serum antibody titer was measured by ELISA. The mRNA expression levels of fatty acid transport-related genes in the adipose tissue of the male broilers were investigated to reveal the relationships among various fatty acid transporters. The results showed that active immunization with FAT/CD36 could significantly decrease the visceral fat of the male broilers by up to 40%, but it had no effect on subcutaneous fat stores of male broilers or on either site of fat deposition in female broilers. The concentration of plasma free fatty acids increased in the experimental groups for both male and female broilers. After the FAT/CD36 immunization, very low density lipoprotein receptor mRNA expression was upregulated in both the subcutaneous and visceral fat of male broilers, whereas peroxisome proliferator-activated receptor γ, FAT/CD36, and acyl-CoA binding protein mRNA expression levels were upregulated only in the visceral fat of male broilers. These results indicated a novel role of chicken FAT/CD36 in fat deposition, with sex- and spatial-specific effects.

Advantages of promoting interleukin-10 by silence of histone deacetylase 11 in inducing tolerance in orthotopic liver transplantation in rats.

AIMS: The aims of this study were to study the role of histone deacetylase 11 (HDAC11) in tolerance induction in orthotopic liver transplantation (OLT) in rats and to assess the advantages of gene therapy over the immunosuppressant FK506. METHODS: Recipients were assigned to an acute rejection group (AcR; group I), an FK506 intervention group (group II), and a tolerance group (group III). Acute rejection (AcR) was graded by the Banff scheme and we examined postoperative survival. The messenger RNA (mRNA) and protein expressions of histone deacetylase 11 (HDAC11) and interleukin (IL) 10 in liver tissue were detected using real-time polymerase chain reaction (PCR) and Western blots, respectively. Plasma levels of tumor necrosis factor (TNF)-α, IL-2, and IL-10 were measured using enzyme-linked immunosorbent Assays. RESULTS: Group I displayed severe, Group II had less, and Group III had no evidence of AR. The survivals among Group III were longer than those in Group I and Group II. IL-10 expression was promoted by HDAC11-shRNA at 7 days after OLT. Serum IL-2 and TNF-α levels were significantly lower among Group III compared with Groups I and II, whereas IL-10 showed the opposite result. CONCLUSIONS: Silence of HDAC11 promotes IL-10 expression and leads to tolerance following OLT in rats. Thus HDAC11 is a promising target for gene therapy to induce tolerance with advantages over immunosuppressive drugs.

Value of imprint cytology for ultrasound-guided transthoracic core biopsy.

The aim of this study was to investigate the possible additional diagnostic information provided by imprint cytology when performing ultrasound-guided transthoracic core biopsy and to evaluate whether it could optimise the biopsy procedure. A total of 155 transthoracic core biopsies with touch imprint smears were performed under ultrasound guidance, with 127 malignant and 28 benign lesions. The imprint smears were stained using Riu's method and interpreted by a cytopathologist. These were compared with the histopathology of core biopsy specimens and the final diagnosis of malignant versus benign disease. The overall diagnostic accuracy of imprint cytology was 94% (146 out of 155). Histopathological analysis showed an overall accuracy of 94% (146 out of 155), with a sensitivity of 94% (119 out of 127) and negative predictive value of 79% (27 out of 34). The combination of these two methodologies had an increased overall accuracy and negative predictive value of 98% (152 out of 155) and 90% (28 out of 31), respectively. The results of imprint cytology and histopathology were in agreement in 143 patients (92%). In conclusion, imprint cytology of ultrasound-guided transthoracic core biopsy is a sensitive procedure for diagnosing peripheral thoracic lesions, and it may increase the diagnostic accuracy and cancer negative prediction of biopsy alone. With an on-site approach, imprint cytology may help to assess the adequacy of biopsy specimens and optimise the biopsy procedure.

Characterization, stability and in-vivo distribution of asialofetuin glycopeptide incorporating DSPC/CHOL liposomes prepared by mild cholate incubation.

In this study, a small triantennary asialoglycopeptide of fetuin (A-F2) was used as a ligand to direct liposomes to hepatocytes. A-F2 was cleaved from asialofetuin, purified, conjugated with fatty acids and incorporated into pre-formed sonicated DSPC/Chol (2:1) liposomes. A mild cholate incubation method for incorporating the A-F2 ligand on pre-formed vesicles was used. In preliminary in vivo experiments 111In3+ encapsulated in A-F2/palmityl liposomes was seen to accumulate in the liver of mice significantly faster than when encapsulated in non-ligand bearing liposomes of the same lipid composition (studied before), justifying further investigation of this system. The presence of the A-F2/fatty acid conjugate in a functional form on the vesicle surface was confirmed by their reversible agglutination in the presence of Ricinus communis agglutinin (RCA120). Effects of ligand incorporation on the vesicle size distribution, z-potential, membrane integrity and stability were monitored. The results demonstrate that highest ligand incorporation was achieved when liposomes and ligand were co-incubated in the presence of 1 mM sodium cholate. Incorporation increased with the length of the fatty acid used for A-F2 conjugation. Ligand-bearing liposomes were demonstrated to be smaller in diameter (about 30%) with a more positive z-potential in comparison to control vesicles while ligand incorporation did not influence the liposome membrane integrity. The size of the ligand-incorporating vesicles was maintained after 24 hours of incubation in isotonic buffer, proving that the vesicles do not aggregate. Although the preliminary biodistribution results may suggest that ligand bearing liposomes are accumulating in the liver, further cell culture, in vivo distribution and especially liver fractionation studies are required in order to clarify the intrahepatic localization of these liposomes and the ability to target liver hepatocytes in vivo.

US-guided transthoracic cutting biopsy for peripheral thoracic lesions less than 3 cm in diameter.

PURPOSE: To evaluate the safety and accuracy of ultrasonography (US)-guided transthoracic cutting biopsy for diagnosing peripheral thoracic lesions (<3 cm). MATERIALS AND METHODS: Fifty consecutive patients with peripheral thoracic lesions less than 3 cm in diameter underwent US-guided percutaneous transthoracic cutting biopsy with a modified technique. Fifty lesions (43 parenchymal lung, two pleural, two chest wall, and three anterior mediastinal lesions) were sampled for biopsy. The final diagnosis was based on histopathologic analysis of surgical specimens (n = 18) or clinical follow-up (n = 32). RESULTS: The histology recovery rate was 98% (49 lesions), and the correct diagnosis was obtained in 48 lesions (96%). Twenty-four (48%) lesions were malignant, and 26 (52%) were benign. The diagnostic accuracy for malignant lesions was 92% (22 of 24 lesions). A specific benign diagnosis was made in 17 (65%) of the 26 benign lesions, and the negative predictive value for malignancy was 93% (26 of 28 lesions). Only two patients (4%) developed postbiopsy pneumothorax, and three (6%) developed postbiopsy hemoptysis. Biopsy helped prevent surgery or thoracoscopy in 32 patients (64%): 18 patients with benign disease and 14 with multiple metastases or inoperable cancer. CONCLUSION: US-guided transthoracic cutting biopsy appears to be a safe and effective method for diagnosing peripheral thoracic lesions less than 3 cm in diameter. The high diagnostic accuracy for benign lesions and metastatic lung cancer can help prevent surgery in many cases.

Bacteriology of infected cavitating lung tumor.

Differentiation between in situ infection and simple tumor necrosis in cavitating lung tumors by means of imaging studies is difficult. In this study, we prospectively investigated the role of ultrasound (US)-guided transthoracic aspiration for bacteriologic examination of infected cavitating lung tumors, and the influence of the culture results on the treatment of patients. Twenty-two patients (18 men and four women) with cavitating lung tumors treated from January 1996 to October 1998 were included. All patients underwent US-guided transthoracic aspiration for bacterial, fungal, and mycobacterial cultures. Microorganisms were isolated from six of seven febrile patients and one of 15 nonfebrile patients. A total of nine pathogens were isolated from seven patients: Klebsiella pneumoniae (n = 3); Haemophilus influenzae (n = 2); Enterococcus faecium (n = 1); Bifidobacterium (n = 1); Shewanella putrefaciens (n = 1); and Mycobacterium tuberculosis (n = 1). Two pathogens were isolated from the aspirate cultures in two patients, while the others had monomicrobial infection. The six febrile patients who had positive lung aspirate cultures were treated with empiric antimicrobial agents before the culture results were available, and the culture results led to adjustment of the antibiotic regimen in five of these. The clinical conditions of the six patients with infected cavitating lung tumors improved after the initiation of individualized antimicrobial treatment. Pneumothorax occurred in one patient, and was the sole procedure-related complication. In conclusion, US-guided transthoracic aspiration is helpful for differentiating infected cavitating lung tumors from simple tumor necrosis. Infection in cavitating lung tumors is common among febrile patients, and the culture results can guide modification of the antimicrobial therapy.

Association analysis of the 5-HT(6) receptor polymorphism (C267T) in mood disorders.

The serotonergic system is implicated in the etiology of mood disorders. Among those most recently discovered serotonin receptors, the relative abundance of serotonin type 6 receptor (5-HT(6)) in the limbic area and the high affinity of some antidepressants to 5-HT(6) receptors suggest that this receptor might be involved in the pathogenesis of mood disorders. In a population-based association study, we tested the hypothesis that the allelic variant (C267T) of the human 5-HT(6) gene confers susceptibility to mood disorders. We genotyped the 5-HT(6) receptor in 139 patients with mood disorders and 147 controls. The results demonstrated that there were no significant differences in genotype or allele frequencies between controls and all patients, or between controls and patients with bipolar disorders or major depression, separately. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 88:601-602, 1999.