Par-4 dependent modulation of cellular ß-catenin by medicinal plant natural product derivative 3-azido Withaferin A.

Here, we provide evidences that natural product derivative 3-azido Withaferin A (3-AWA) abrogated EMT and invasion by modulating ß-catenin localization and its transcriptional activity in the prostate as well as in breast cancer cells. This study, for the first time, reveals 3-AWA treatment consistently sequestered nuclear ß-catenin and augmented its cytoplasmic pool as evidenced by reducing ß-catenin transcriptional activity in these cells. Moreover, 3-AWA treatment triggered robust induction of pro-apoptotic intracellular Par-4, attenuated Akt activity and rescued Phospho-GSK3ß (by Akt) to promote ß-catenin destabilization. Further, our in vitro studies demonstrate that 3-AWA treatment amplified E-cadherin expression along with sharp downregulation of c-Myc and cyclin D1 proteins. Strikingly, endogenous Par-4 knock down by siRNA underscored 3-AWA mediated inhibition of nuclear ß-catenin was Par-4 dependent and suppression of Par-4 activity, either by Bcl-2 or by Ras transfection, restored the nuclear ß-catenin level suggesting Par-4 mediated ß-catenin regulation was not promiscuous. In vivo results further demonstrated that 3-AWA was effective inhibitor of tumor growth and immunohistochemical studies indicated that increased expression of total ß-catenin and decreased expression of phospho-ß-catenin and Par-4 in breast cancer tissues as compared to normal breast tissue suggesting Par-4 and ß-catenin proteins are mutually regulated and inversely co-related in normal as well as cancer condition. Thus, strategic regulation of intracellular Par-4 by 3-AWA in diverse cancers could be an effective tool to control cancer cell metastasis. Conclusively, this report puts forward a novel approach of controlling deregulated ß-catenin signaling by 3-AWA induced Par-4 protein.

Incidence of pseudotumor and acute lymphocytic vasculitis associated lesion (ALVAL) reactions in metal-on-metal hip articulations: a meta-analysis.

We systematically reviewed the peer-reviewed literature to determine a pooled estimate of the incidence of pseudotumor and acute lymphocytic vasculitis associated lesions (ALVAL) in adult patients with primary metal-on-metal (MoM) total hip arthroplasty or resurfacing. Fourteen eligible articles were identified, with a total of 13,898 MoM hips. The incidence of pseudotumor/ALVAL ranged from 0% to 6.5% of hips with a mean follow-up ranging from 1.7 to 12.3 years across the studies. The pooled estimated incidence of pseudotumor/ALVAL is 0.6% (95% CI: 0.3% to 1.2%). The rate of revision for any reason ranged from 0% to 14.3% of hips, with a pooled estimate of 3.9% (95% CI: 2.7% to 5.3%).

Dual modulation of Ras-Mnk and PI3K-AKT-mTOR pathways: A Novel c-FLIP inhibitory mechanism of 3-AWA mediated translational attenuation through dephosphorylation of eIF4E.

The eukaryotic translation initiation factor 4E (eIF4E) is considered as a key survival protein involved in cell cycle progression, transformation and apoptosis resistance. Herein, we demonstrate that medicinal plant derivative 3-AWA (from Withaferin A) suppressed the proliferation and metastasis of CaP cells through abrogation of eIF4E activation and expression via c-FLIP dependent mechanism. This translational attenuation prevents the de novo synthesis of major players of metastatic cascades viz. c-FLIP, c-Myc and cyclin D1. Moreover, the suppression of c-FLIP due to inhibition of translation initiation complex by 3-AWA enhanced FAS trafficking, BID and caspase 8 cleavage. Further ectopically restored c-Myc and GFP-HRas mediated activation of eIF4E was reduced by 3-AWA in transformed NIH3T3 cells. Detailed underlying mechanisms revealed that 3-AWA inhibited Ras-Mnk and PI3-AKT-mTOR, two major pathways through which eIF4E converges upon eIF4F hub. In addition to in vitro studies, we confirmed that 3-AWA efficiently suppressed tumor growth and metastasis in different mouse models. Given that 3-AWA inhibits c-FLIP through abrogation of translation initiation by co-targeting mTOR and Mnk-eIF4E, it (3-AWA) can be exploited as a lead pharmacophore for promising anti-cancer therapeutic development.

Billing and coding knowledge: a comparative survey of professional coders, practicing orthopedic surgeons, and orthopedic residents.

Medical knowledge and surgical skills are necessary to become an effective orthopedic surgeon. To run an efficient practice, the surgeon must also possess a basic understanding of medical business practices, including billing and coding. In this study, we surveyed and compared the level of billing and coding knowledge among current orthopedic residents PGY3 and higher, academic and private practice attending orthopedic surgeons, and orthopedic coding professionals. According to the survey results, residents and fellows have a similar knowledge of coding and billing, regardless of their level of training or type of business education received in residency. Most residents would like formal training in coding, billing, and practice management didactics; this is consistent with data from previous studies.

A novel MMP-2 inhibitor 3-azidowithaferin A (3-azidoWA) abrogates cancer cell invasion and angiogenesis by modulating extracellular Par-4.

BACKGROUND: Withaferin A, which is a naturally derived steroidal lactone, has been found to prevent angiogenesis and metastasis in diverse tumor models. It has also been recognized by different groups for prominent anti-carcinogenic roles. However, in spite of these studies on withanolides, their detailed anti-metastatic mechanism of action remained unknown. The current study has poised to address the machinery involved in invasion regulation by stable derivative of Withaferin A, 3-azido Withaferin A (3-azidoWA) in human cervical HeLa and prostate PC-3 cells. METHODS AND PRINCIPAL FINDINGS: Sub-toxic concentration of 3-azidowithaferin A (3-azido WA) inhibited cancer cell motility and invasion in wound healing and Boyden chamber invasion by suppressing MMP-2 activity in gelatin zymography and its expression has proved to be a major obstacle in chemo-sensitivity. We have uncovered a novel mechanism of 3-azidoWA induced extracellular pro-apoptotic candidate tumor suppressor Par-4 protein stimulation in conditioned media and also noticed a concomitant marked reduction in pAkt and pERK signaling by immunoblot analysis. Furthermore, our zymography results suggest 3-azidoWA induced MMP-2 inhibition was mediated through secretory Par-4. The inhibition of apoptosis by 3-azidoWA could not restore MMP-2 gelatinase activity. In addition to this, our in vivo animal experiments data showed 3-azidoWA abrogated neovascularisation in dose dependent manner in mouse Matrigel plug assay. CONCLUSION/SIGNIFICANCE: For this report, we found that 3-azidoWA suppressed motility and invasion of HeLa and PC-3 cells in MMP-2 dependent manner. Our in vitro result strongly suggests that sub-toxic doses of 3-azidoWA enhanced the secretion of extracellular Par-4 that abolished secretory MMP-2 expression and activity. Depletion of secretory Par-4 restored MMP-2 expression and invasion capability of HeLa and PC-3 cells. Further, our findings implied that 3-azidoWA attenuated internal phospho-ERK and phospho-Akt expression in a dose dependent manner might play a key role in inhibition of mouse angiogenesis by 3-azidoWA.

Static load repetition is a risk factor in the development of lumbar cumulative musculoskeletal disorder.

STUDY DESIGN: In vivo feline model subjected to variable number of repetitions of a short static lumbar flexion followed by an equally long rest period. OBJECTIVES: The purpose of this study was to determine the influence of the number of repetitions as a risk factor in promoting a cumulative low back disorder in the feline model. SUMMARY OF BACKGROUND DATA: Epidemiologic data point out that the increased number of repetitions of static lumbar loading is a major risk factor in the development of cumulative low back disorder. Biomechanical and physiologic confirmation of the epidemiology is lacking. Recent work demonstrated that repetitive static loading results in accumulation of creep in the lumbar viscoelastic tissues, resulting in a neuromuscular disorder consisting of spasms during loading and hyperexcitability of lumbar muscles during following rest. It was also shown that the load magnitude is a major risk factor. It is hypothesized that increased number of repetitions of static load periods result in increased severity of the resulting neuromuscular disorder. METHODS: Static lumbar flexion of 10 minutes duration followed by 10 minutes rest was repeated three times in one experimental group, six times in the second, and nine times in the third group. In all groups, the creep developing in the lumbar viscoelastic tissues as well as the reflexive EMG from the multifidus were monitored during the flexion/rest periods and throughout a 7-hour recovery period after the repetitions. RESULTS: Creep developed and accumulated during each of the flexion/rest periods in the three experimental protocols, with larger residual creep at the end of the nine repetitions. A residual creep was still present at the end of the 7 hours of recovery allowed in each of the three groups. During the flexion/rest sessions, EMG spasms were present, and the presence of an initial hyperexcitability was detected during the 7 hours of recovery in all the preparations. The presence of a delayed hyperexcitability was revealed only in the group subjected to nine flexion/rest periods, while it was not observed in the groups subjected to three and six flexion/rest repetitions. The statistical analysis (post hoc Fisher test) performed on the normalized integrated EMG and displacement data during the recovery phase showed a significant difference between the nine repetitions group and the other two groups (P < 0.0001). The two-way ANOVA analysis revealed a significant effect of time (P < 0.005) and number of repetitions (P < 0.0001) on all considered parameters. CONCLUSIONS: It was concluded that a cumulative neuromuscular disorder develops because of repetition of static lumbar flexion, and the severity of the disorder provoked is magnified by the number of repetitions. Despite the highly favorable 1:1 work-to-rest ratio and the 7-hour post loading rest period, a full recovery of creep was not obtained in this study.