Osteogenesis induced in goat bone marrow progenitor cells by recombinant adenovirus coexpressing bone morphogenetic protein 2 and basic fibroblast growth factor

Bone morphogenetic protein 2 (BMP2) and basic fibroblast growth factor (bFGF) have been shown to exhibit a synergistic effect to promote bone repair and healing. In this study, we constructed a novel adenovirus with high coexpression of BMP2 and bFGF and evaluated its effect on osteogenic differentiation of goat bone marrow progenitor cells (BMPCs). Recombinant adenovirus Ad-BMP2-bFGF was constructed by using the T2A sequence. BMPCs were isolated from goats by density gradient centrifugation and adherent cell culture, and were then infected with Ad-BMP2-bFGF or Ad-BMP2. Expression of BMP2 and bFGF was detected by ELISA, and alkaline phosphatase (ALP) activity was detected by an ALP assay kit. In addition, von Kossa staining and immunocytochemical staining of collagen II were performed on BMPCs 21 days after infection. There was a high coexpression of BMP2 and bFGF in BMPCs infected with Ad-BMP2-bFGF. Twenty-one days after infection, ALP activity was significantly higher in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. Larger and more mineralized calcium nodules, as well as stronger collagen II staining, were observed in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. In summary, we developed a novel adenovirus vector Ad-BMP2-bFGF for simultaneous high coexpression of BMP2 and bFGF, which could induce BMPCs to differentiate efficiently into osteoblasts.

Expression of beclin 1 in primary salivary adenoid cystic carcinoma and its relation to Bcl-2 and p53 and prognosis

Beclin 1 plays a critical role in autophagy and functions as a haploinsufficient tumor suppressor. The expression and prognostic significance of beclin 1 in head and neck adenoid cystic carcinoma (ACC) are largely unexplored. Therefore, we investigated the expression of beclin 1, Bcl-2, and p53 in head and neck ACC tissue. Tissue samples from 35 cases (15 females, 20 males) of head and neck ACC were utilized for immunohistochemistry. Beclin 1 expression was observed in 32 cases (91.4%) and considered to be high in 15 cases (42.9%) and low in 20 cases (57.1%). Beclin 1 expression was significantly correlated with a histological growth pattern (P=0.046) and histological grade (P=0.037). Beclin 1 expression was inversely correlated with Bcl-2 expression (P=0.013) and significantly associated with overall survival (P=0.006). Bcl-2 and p53 expression were observed in 21 cases (60.0%) and 16 cases (45.7%). Bcl-2 expression was significantly correlated with perineural invasion (P=0.041) and not associated with overall survival (P=0.053). p53 expression was directly correlated with beclin 1 expression (P=0.044). Our results indicated that beclin 1 may be a novel, promising prognostic factor for clinical outcome in head and neck ACC patients and may play a part in the development of head and neck ACC by interacting with Bcl-2 and p53.

BMP-2 and titanium particles synergistically activate osteoclast formation

A previous study showed that BMP-2 (bone morphogenetic protein-2) and wear debris can separately support osteoclast formation induced by the receptor activator of NF-κB ligand (RANKL). However, the effect of BMP-2 on wear debris-induced osteoclast formation is unclear. In this study, we show that neither titanium particles nor BMP-2 can induce osteoclast formation in RAW 264.7 mouse leukemic monocyte macrophage cells but that BMP-2 synergizes with titanium particles to enhance osteoclast formation in the presence of RANKL, and that at a low concentration, BMP-2 has an optimal effect to stimulate the size and number of multinuclear osteoclasts, expression of osteoclast genes, and resorption area. Our data also clarify that the effects caused by the increase in BMP-2 on phosphorylated SMAD levels such as c-Fos expression increased throughout the early stages of osteoclastogenesis. BMP-2 and titanium particles stimulate the expression of p-JNK, p-P38, p-IkB, and P50 compared with the titanium group. These data suggested that BMP-2 may be a crucial factor in titanium particle-mediated osteoclast formation.

Unusual association of NDM-1 with KPC-2 and armA among Brazilian Enterobacteriaceae isolates

We report the microbiological characterization of four New Delhi metallo-β-lactamase-1 (blaNDM-1)-producing Enterobacteriaceae isolated in Rio de Janeiro, Brazil. blaNDM-1 was located on a conjugative plasmid and was associated with Klebsiella pneumoniae carbapenemase-2 (blaKPC-2) or aminoglycoside-resistance methylase (armA), a 16S rRNA methylase not previously reported in Brazil, in two distinct strains of Enterobacter cloacae. Our results suggested that the introduction of blaNDM-1 in Brazil has been accompanied by rapid spread, since our isolates showed no genetic relationship.

The roles of ERK1/2 and PI3K in abnormal vascular functions in angiotensin II-infused hypertensive rats

Background: Ang II plays a major role in cardiovascular regulation. Recently, it has become apparent that vascular superoxide anion may play an important role in hypertension development. Treatment with antisense NAD(P)H oxidase or SOD decreased BP in Ang II-infused rats. Wang et al recently reported mice which lack one of the subunits of NAD(P)H oxidase developed hypertension at a much lower extent when compared to the wild type animals infused with Ang II, indicating that superoxide anion contributes to elevation in BP in the Ang II-infused hypertensive model. In the Ang II-infused hypertensive model, altered reactivity of blood vessels is often associated with the elevation of systolic blood pressure. We have observed abnormal tension development and impaired endothelium-dependent relaxation in the isolated aorta of Ang II-infused and DOCA-salt hypertensive rats. Recently, several other cellular signal molecules, including ERK1I2 and PI3K, have been determined to play important roles in the regulation of smooth muscle contraction and relaxation. ERKl/2 and PI3K pathways are also reported to contribute to Ang II induced cell growth, hypertrophy, remodeling and contraction. Moreover, these signaling pathways have shown ROS-sensitive properties. Therefore, the aim of the present study is to investigate the roles of ERKl12 and PI3K in vascular oxidative stress, spontaneous tone and impaired endothelium relaxation in Ang II-infused hypertensive model. Hypothesis: We hypothesize that the activation of ERKl12 and PI3K are elevated in response to an Ang II infusion for 6 days. The elevated activation of phospho-ERKl/2 and PI3K mediated the increased level of vascular superoxide anion, the abnormal vascular contraction and impaired endothelium-dependent vascular relaxation in Ang II-infused hypertensive rats. Methods: Vascular superoxide anion level is measured by lucigenin chemiluminescence. Spontaneous tone and ACh-induced endothelium-dependent relaxation was measured by isometric tension recording in organ chamber. The activity of ERK pathway will be measured by its Western blot of phosphorylation of ERK. PI3K activity was evaluated indirectly by Western blot of the phosphorylation of PDKl, a downstream protein of PI3K signaling pathway. The role of each pathway was also addressed via comparing the responses to the specific inhibitors. Results: Superoxide anion was markedly increased in the isolated thoracic aorta from Ang II-infused rats. There was spontaneous tone developed in rings from Ang II-induced hypertensive but not sham-operated normotensive rats. ACh-induced endothelium-dependent relaxation function is impaired in Ang II-infused hypertensive rats. Superoxide dismutase and NAD(P)H oxidase inhibitor, apocynin, inhibited the abnormal spontaneous tone and ameliorated impaired endothelium-dependent relaxation. The expression of phopho-ERKII2 was enhanced in Ang II-infused rats, indicating the activity of ERK1I2 could be increased. MEK1I2 inhibitors, PD98059 and U126, but not their inactive analogues, SB203580 and U124, significantly reduced the vascular superoxide anion in aortas from Ang II-infused rats. The MEK1I2 inhibitors reduced the spontaneous tone and improved the impaired endothelium-dependent relaxation in aorta of hypertension. These findings supported the role of ERKII2 signaling pathway in vascular oxidative stress, spontaneous tone and impaired endothelium-dependent relaxation in Ang II-infused hypertensive rats. The amount of phospho-PDK, a downstream protein of PI3K was increased in Ang II rats indicating the activity of PI3K activity was elevated. Strikingly, PI3K significantly inhibited the increase of superoxide anion level, abnormal spontaneous tone and restored endothelium-dependent relaxation in Ang II-infused hypertensive rats. These findings indicated the important role of PI3K in Ang II-infused hypertensive rats. Conclusion: ERKII2 and PI3K signaling pathways are sustained activated in Ang II-infused hypertensive rats. The activated ERKII2 and PI3K mediate the increase of vascular superoxide anion level, vascular abnormal spontaneous tone and impaired endothelium-dependent relaxation.

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month of Sept. 1854, signed by S.D. Woodruff, Oct. 1854.

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines during the month ending Oct. 31, 1854.

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month of Oct. 1854

Estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month of Oct. 1854, signed by S.D. Woodruff, Nov. 1854.

Chart of estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month ending Dec. 1, 1854

Chart of estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month ending Dec. 1, 1854, signed by S.D. Woodruff, Jan. 1855.

An estimate of work done by Messrs. Brown and McDonell, contractors on sections 1, 2 and 3 ending at St. Catharines for the months of April and May 1855

Scrap of paper that has been burned almost completely. Only the header of the document is intact. It is an estimate of work done by Messrs. Brown and McDonell, contractors on sections 1, 2 and 3 ending at St. Catharines for the months of April and May 1855.

Chart of estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines

Chart of estimate of work done on the Port Dalhousie and Thorold Railway by Messrs. Brown and McDonell, contractors, on sections 1, 2, and 3 ending at St. Catharines for the month of Oct., 1855. This document is water damaged and full of holes. The list of prices is almost completely illegible. The document is signed by S.D. Woodruff, Nov. 1855.

Schedule of prices for Brown and McDonell, contractors, for sections 1, 2 and 3 of the Port Dalhousie Thorold Railway

Schedule of prices for Brown and McDonell, contractors, for sections 1, 2 and 3 of the Port Dalhousie Thorold Railway (1 page, handwritten), Sept. 24, 1856.

County of Welland estimate (copy) of work done on sections 2 and 3 of the tap drain at Marshville by George Stanford

County of Welland estimate (copy) of work done on sections 2 and 3 of the tap drain at Marshville by George Stanford, signed by S.D. Woodruff. Estimate no.1, July, 1856.

County of Welland estimate (copy) of work done on sections 2 and 3 of the tap drain at Marshville by George Stanford

County of Welland estimate (copy) of work done on sections 2 and 3 of the tap drain at Marshville by George Stanford, signed by S.D. Woodruff. Estimate no.2, Oct., 1856.

Chart of land drainage for the Welland Canal final estimate of work done on sections no.1, 2 and 3

Chart of land drainage for the Welland Canal final estimate of work done on sections no.1, 2 and 3 on the road below lock no. 2 leading to Port Dalhousie. Work commenced Nov. 1846 and finished July 1847. Road work and the waste weir no.1 to Port Dalhousie work commenced Aug. 1847 and finished Sept. 1847, Nov.1, 1847.