Dynamin Is Functionally Coupled to Insulin Granule Exocytosis

The insulin granule integral membrane protein marker phogrin-green fluorescent protein was co-localized with insulin in Min6B1 beta-cell secretory granules but did not undergo plasma membrane translocation following glucose stimulation. Surprisingly, although expression of a dominant-interfering dynamin mutant (Dyn/K44A) inhibited transferrin receptor endocytosis, it had no effect on phogringreen fluorescent protein localization in the basal or secretagogue-stimulated state. By contrast, co-expression of Dyn/K44A with human growth hormone as an insulin secretory marker resulted in a marked inhibition of human growth hormone release by glucose, KCl, and a combination of multiple secretagogues. Moreover, serial pulse depolarization stimulated an increase in cell surface capacitance that was also blocked in cells expressing Dyn/K44A. Similarly, small interference RNA-mediated knockdown of dynamin resulted in marked inhibition of glucose-stimulated insulin secretion. Together, these data suggest the presence of a selective kiss and run mechanism of insulin release. Moreover, these data indicate a coupling between endocytosis and exocytosis in the regulation of beta-cell insulin secretion

Improvement of the Photoelectrochemical Stability of Cu2O Photocathode by Ph—CΞC—Cu Grafting

As one of the most efficient photocathodes, Cu2O has attracted substantial attention because of its theoretically high solar-to-hydrogen (STH) efficiency. However, its applications in photoelectrochemical (PEC) fields are severely restricted by the poor stability derived from serious photocorrosion. In this work, high-quality phenylethynyl copper (Ph—CΞC—Cu) are successfully self-assembled on the surface of Cu2O photocathode by a novel photothermal method to improve its photostability. With the protection of the Ph—CΞC—Cu layer, 85% of the initial photocurrent density can be remained, while only 28% of initial photocurrent density is left on bare Cu2O photocathode prepared on a copper foam (CF) substrate. The significantly improved photostability of Cu2O photocathode by the Ph—CΞC—Cu protective layer is attributed to its strong hydrophobicity, which can efficiently inhibit the corrosion of Cu2O by the aqueous electrolyte solution due to its special crystal structure. Based on the obtained Ph—CΞC—Cu/Cu2O photocathode, a two-photoelectrode cell with excellent stability (>5 h) has also been successfully constructed for water splitting without the need of an electric bias

Giant schwannoma of thoracic vertebra: A case report

BACKGROUND,It is relatively rare for schwannomas to invade bone, but it is very rare for a large,mass to form concurrently in the paravertebral region. Surgical resection is the,only effective treatment. Because of the extensive tumor involvement and the,many important surrounding structures, the tumor needs to be fully exposed.,Most of the tumors are completely removed by posterior combined open-heart,surgery to relieve spinal cord compression, restore the stability of the spine and,maximize the recovery of nerve and spinal cord function. The main objective of,this article is to present a schwannoma that had invaded the T5 and T6 vertebral,bodies and formed a large paravertebral mass with simultaneous invasion of the,spinal canal and compression of the spinal cord.,CASE SUMMARY,A 40-year-old female suffered from intermittent chest and back pain for 8 years.,Computed tomography and magnetic resonance imaging scans showed a,paravertebral tumor of approximately 86 mm × 109 mm × 116 mm, where the,adjacent T5 and T6 vertebral bodies were invaded by the tumor, the right intervertebral,foramen was enlarged, and the tumor had invaded the spinal canal to,compress the thoracic medulla. The preoperative puncture biopsy diagnosed a,benign schwannoma. Complete resection of the tumor was achieved by a two-step,operation. In the first step, the thoracic surgeon adopted a lateral approach to,separate the thoracic tumor from the lung. In the second step, a spine surgeon,performed a posterior midline approach to dissect the tumor from the vertebral,junction through removal of the tumor from the posterior side and further,resection of the entire T5 and T6 vertebral bodies. The large bone defect was,reconstructed with titanium mesh, and the posterior root arch was nail-fixed. Due,to the large amount of intraoperative bleeding, we performed tumor angioembolization,before surgery to reduce and avoid large intraoperative bleeding. The,postoperative diagnosis of benign schwannoma was confirmed by histochemical,examination. There was no sign of tumor recurrence or spinal instability during,the 2-year follow-up.,CONCLUSION,Giant schwannoma is uncommon. In this case, a complete surgical resection of a,giant thoracic nerve sheath tumor that invaded part of the vertebral body and,compressed the spinal cord was safe and effective

Infection with hepatitis B virus carrying novel pre-S/S gene mutations in female siblings vaccinated at birth: two case reports

<p>Abstract</p> <p>Introduction</p> <p>After the initiation of a mass hepatitis B vaccination program in Taiwan, the prevalence of hepatitis B virus infection has declined progressively. However, about 1 percent of the young generation, who received hepatitis B vaccination at birth, remain carriers. Infection with vaccine-escape hepatitis B virus mutants always occurs shortly after birth. Here, we report two female siblings in whom the infection occurred in their adolescence. This report raises the question of whether a booster for hepatitis B vaccination is needed.</p> <p>Case presentation</p> <p>Two 19 and 14-year-old Taiwanese female siblings were born to a mother infected with hepatitis B virus and received a complete course of hepatitis B vaccination at birth. They remained negative for serum hepatitis B surface antigen and positive for serum anti-hepatitis B surface antibody throughout their childhood. However, both were infected with the hepatitis B virus in their adolescence. Hepatitis B virus DNA was extracted from serum samples from the mother and two siblings. Hepatitis B virus pre-S/S sequence was amplified by polymerase chain reaction followed by nucleotide sequencing. When compared with the sequence obtained from the mother, multiple amino acid substitutions located near or in the major hydrophilic region of the surface antigen were identified in the elder sister. Four of these mutations (sL97S, sL98S, sG102R, and sA159P) were novel. A novel in-frame deletion (14 amino acids deleted, pre-S 127-140) was found in the hepatitis B virus pre-S2 region in the younger sister.</p> <p>Conclusions</p> <p>Despite having received hepatitis B vaccination at birth, hepatitis B virus infection can still occur in adolescence with the emergence of novel mutations in the pre-S/S gene. This is a rare event and, to the best of our knowledge, has not been previously reported.</p

The impact of shadow banking on the implementation of Chinese monetary policy

This paper empirically analyses the relationship between the shadow banking system and implementation of monetary policy in China using the VECM methodology. We show that an increase in the size of shadow banking sector increases the independence of bank lending from the policies of the People Bank of China. We also find that Shadow Banking works in an asymmetric fashion in that it amplifies increases in the money supply but weakens the effects of restrictive interest rate-based monetary policy decisions

Interleukin-17D and Nrf2 mediate initial innate immune cell recruitment and restrict MCMV infection.

Innate immune cells quickly infiltrate the site of pathogen entry and not only stave off infection but also initiate antigen presentation and promote adaptive immunity. The recruitment of innate leukocytes has been well studied in the context of extracellular bacterial and fungal infection but less during viral infections. We have recently shown that the understudied cytokine Interleukin (IL)-17D can mediate neutrophil, natural killer (NK) cell and monocyte infiltration in sterile inflammation and cancer. Herein, we show that early immune cell accumulation at the peritoneal site of infection by mouse cytomegalovirus (MCMV) is mediated by IL-17D. Mice deficient in IL-17D or the transcription factor Nuclear factor (erythroid-derived 2)-like 2 (Nrf2), an inducer of IL-17D, featured an early decreased number of innate immune cells at the point of viral entry and were more susceptible to MCMV infection. Interestingly, we were able to artificially induce innate leukocyte infiltration by applying the Nrf2 activator tert-butylhydroquinone (tBHQ), which rendered mice less susceptible to MCMV infection. Our results implicate the Nrf2/IL-17D axis as a sensor of viral infection and suggest therapeutic benefit in boosting this pathway to promote innate antiviral responses

Expression, Localization, and Phosphorylation of Akt1 in Benign and Malignant Thyroid Lesions

The serine/threonine protein kinase Akt is a key molecule in the phosphatidyl inositol 3-kinase pathway that is often overactivated in human cancers. Three Akt isoforms (Akt1, Akt2, Akt3) have been identified in human cells and they show different distribution and have non-redundant functions. The aim of this study was to determine whether the expression, phosphorylation, and localization of Akt1 isoform in human thyroid malignant lesions are different from those in benign lesions. Nuclear and cytoplasmic fractions were isolated from tissue samples and Western blot method was used to detect Akt1 presence in both cellular fractions. Akt1 expression was also assessed by ELISA method. To estimate Akt1 phosphorylation, kinase was immunoprecipitated from cell lysates and tested with anti-phospho-Akt antibodies. The Akt1 expression in majority of thyroid cancer samples was significantly higher than in benign lesions (p < 0.05). Akt1 both in differentiated cancers (follicular and papillary) and benign lesions was localized mainly in cytoplasmic fraction. In two of three anaplastic cancer samples Akt1 was predominantly localized in nucleus. The ratio of phosphorylated Akt1 to total Akt1 was lower in cancers than in non-neoplastic lesions and adenomas. Thus, although Akt1 seems to be overexpressed in thyroid neoplasms, its high phosphorylation is not characteristic for thyroid cancers

Asiatic Acid Inhibits Liver Fibrosis by Blocking TGF-beta/Smad Signaling In Vivo and In Vitro

Liver fibrosis is a major cause of liver failure, but treatment remains ineffective. In the present study, we investigated the mechanisms and anti-hepatofibrotic activities of asiatic acid (AA) in a rat model of liver fibrosis induced by carbon tetrachloride (CCl4) and in vitro in TGF-beta1-stimulated rat hepatic stellate cell line (HSC-T6). Treatment with AA significantly attenuated CCl4-induced liver fibrosis and functional impairment in a dosage-dependent manner, including blockade of the activation of HSC as determined by inhibiting de novo alpha smooth muscle actin (a-SMA) and collagen matrix expression, and an increase in ALT and AST (all p<0.01). The hepatoprotective effects of AA on fibrosis were associated with upregulation of hepatic Smad7, an inhibitor of TGF-beta signaling, thereby blocking upregulation of TGF-beta1 and CTGF and the activation of TGF-beta/Smad signaling. The anti-fibrosis activity and mechanisms of AA were further detected in vitro in HSC-T6. Addition of AA significantly induced Smad7 expression by HSC-T6 cells, thereby inhibiting TGF-beta1-induced Smad2/3 activation, myofibroblast transformation, and collagen matrix expression in a dosage-dependent manner. In contrast, knockdown of Smad7 in HSC-T6 cells prevented AA-induced inhibition of HSC-T6 cell activation and fibrosis in response to TGF-beta1, revealing an essential role for Smad7 in AA-induced anti-fibrotic activities during liver fibrosis in vivo and in vitro. In conclusion, AA may be a novel therapeutic agent for liver fibrosis. Induction of Smad7-dependent inhibition of TGF-beta/Smad-mediated fibrogenesis may be a central mechanism by which AA protects liver from injury

In Situ Monitoring of Intracellular Glucose and Glutamine in CHO Cell Culture

The development of processes to produce biopharmaceuticals industrially is still largely empirical and relies on optimizing both medium formulation and cell line in a product-specific manner. Current small-scale (well plate-based) process development methods cannot provide sufficient sample volume for analysis, to obtain information on nutrient utilization which can be problematic when processes are scaled to industrial fermenters. We envision a platform where essential metabolites can be monitored non-invasively and in real time in an ultra-low volume assay in order to provide additional information on cellular metabolism in high throughput screens. Towards this end, we have developed a model system of Chinese Hamster Ovary cells stably expressing protein-based biosensors for glucose and glutamine. Herein, we demonstrate that these can accurately reflect changing intracellular metabolite concentrations in vivo during batch and fed-batch culture of CHO cells. The ability to monitor intracellular depletion of essential nutrients in high throughput will allow rapid development of improved bioprocesses

Evaluation of macrophage migration inhibitory factor as an imaging marker for hepatocellular carcinoma in murine models

Objective. Macrophage migration inhibitory factor (MIF) is considered as an important mediator in the pathogenesis of neoplasia. The aim of the present study was to evaluate whether MIF could be used as a marker for hepatocellular carcinoma (HCC) detection. Material and methods. Biodistribution and whole-body autoradiography studies of 131I-labeled anti-MIF monoclonal antibody (McAb) and 131I-labeled control IgG were performed. The HCC-bearing mice were injected with 3.7 MBq of each agent and killed at 24, 48, and 72 h postinjection (p.i.). The organs, blood, and HCC tissues were removed from model mice, weighed, and counted using a gamma-counter. The expression of MIF mRNA and protein within HCC tissues was confirmed by RT-PCR and immunohistochemistry. Results. HCCs in model mice could be adequately visualized at 24 h p.i. The target-to-non-target (T/NT) ratios were 6.72 ± 1.09 (24 h), 9.85 ± 0.81 (48 h), and 12.31 ± 0.57 (72 h) for 131I-labeled anti-MIF McAb group, whereas in the control group of 131I-IgG, T/NT ratios were 4.65 ± 0.63 (24 h), 6.12 ± 0.60 (48 h), and 8.23 ± 0.35 (72 h) (p < 0.05). MIF mRNA expression was twofold higher in the HCC tissues than in the healthy liver tissues. MIF protein expression was much higher in the HCC tissues than in controls. Conclusions. Our findings suggested that 131I-anti-MIF McAb could be rapidly and specifically localized in tumors. Thus, MIF could be used as a marker for HCC tumor detection