Early detection of metastasis can be aided by circulating tumor cells (CTCs), which also show potential to predict early relapse. Due to the limited CTC numbers in peripheral blood in early stages, we investigated CTCs in pulmonary vein blood accessed during surgical resection of tumors. Pulmonary vein (PV) and peripheral vein (Pe) blood specimens from patients with lung cancer were drawn during the perioperative period and assessed for CTC burden using a microfluidic device. From 108 blood samples analyzed from 36 patients, PV had significantly higher number of CTCs compared to pre-operative Pe (p<0.0001) and intra-operative Pe (p<0.001) blood. CTC clusters with large number of CTCs were observed in 50% of patients, with PV often revealing larger clusters. Long term surveillance indicated that presence of clusters in pre-operative Pe blood predicted a trend toward poor prognosis. Gene expression analysis by RT-qPCR revealed enrichment of p53 signaling and extracellular matrix involvement in PV and Pe samples. Ki67 expression was detected in 62.5% of PV samples and 59.2% of Pe samples, with the majority (72.7%) of patients positive for Ki67 expression in PV having single CTCs as opposed to clusters. Gene ontology analysis revealed enrichment of cell migration and immune-related pathways in CTC clusters, suggesting survival advantage of clusters in circulation. Clusters display characteristics of therapeutic resistance, indicating the aggressive nature of these cells. Thus, CTCs isolated from early stages of lung cancer are predictive of poor prognosis and can be interrogated to determine biomarkers predictive of recurrence.
Circulating tumor cells (CTCs) are believed to play an important role in metastasis, a process responsible for the majority of cancer-related deaths. But their rarity in the bloodstream makes microfluidic isolation complex and time-consuming. Additionally the low processing speeds can be a hindrance to obtaining higher yields of CTCs, limiting their potential use as biomarkers for early diagnosis. Here we report a high throughput microfluidic technology, the OncoBean Chip, employing radial flow that introduces a varying shear profile across the device enabling efficient cell capture by affinity at high flow rates. The recovery from whole blood was validated with cancer cell lines H1650 and MCF7, achieving a mean efficiency >80% at a throughput of 10 mL hr−1 in contrast to a flow rate of 1 mL hr−1 standardly reported with other microfluidic devices. Cells were recovered with a viability rate of 93% at these high speeds, increasing the ability to use captured CTCs for downstream analysis. Broad clinical application was demonstrated using comparable flow rates from blood specimens obtained from breast, pancreatic and lung cancer patients. Comparable CTC numbers were recovered in all the samples at the two flow rates demonstrating the ability of the technology to perform at high-throughputs.
Introduction: In a populous city like Mumbai, which lacks an organized prehospital emergency medical services (EMS) system, there exists an informal network through which victims arrive at the trauma center. This baseline study describes the prehospital care and transportation that currently is available in Mumbai. Methods: A prospective trauma database was created by interviewing 170 randomly selected patients from a total of 454 admitted over a two-month period (July-August 2005) at a Level-I, urban, trauma center. Results: The injured victim in Mumbai usually is rescued by a good Samaritan passer-by (43.5%) and contrary to popular belief, helped by the police (89.7%). Almost immediately after rescue, the victim begins transport to the hospital. No one waits for the EMS ambulance to arrive, as there is none. A taxi cab is the most popular substitute for the ambulance (39.3%). The trauma patient in India usually is a young man in his late-twenties, from a lower socioeconomic class. He mostly finds himself in a government hospital, as private hospitals are reluctant to provide trauma care to the seriously injured. The injured who do receive prehospital care receive inadequate and inappropriate care due to the high cost of consumables in resuscitation, and in part due to the providers' lack of training in emergency care. Those who were more likely to receive prehospital care suffered from road traffic injuries (odds ratio (OR) = 2.3) and those transported by government ambulances (OR = 10.83), as compared to railway accident victims (OR = 0 .41) and those who came by taxi (OR = 0.54). Conclusions: Currendy, as a result of not having an EMS system, prehospital care is a citizen responsibility using societal networks. It is easy to eliminate this system and shift the responsibility to the state. The moot point is whether the state-funded EMS system will be robust enough in a resource-poor setting in which public hospitals are poorly funded. Considering the high funding cost of EMS systems in developed countries and the insufficient evidence that prehospital field interventions by the EMS actually have improved outcomes, Mumbai must proceed with caution when implementing advanced EMS systems into its congested urban traffic. Similar cities, such as Mexico City and Jakarta, have had limited success with implementing EMS systems. Perhaps reinforcing the existing network of informal providers of taxi drivers and police and with training, funding quick transport with taxes on roads and automobile fuels and regulating the private ambulance providers, could be more cost-effective in a culture in which sharing and helping others is not just desirable, but is necessary for overall economic survival. Roy N, Murlidhar V, Chowdhury R, Patil SB, Supe PA, Vaishnav PD, Vatkar A: Where there are no emergency medical services-Prehospital care for the injured in Mumbai, India. Prehosp D«
The study of circulating tumor cells (CTCs) has been made possible by many technological advances in their isolation. Their isolation has seen many fronts, but each technology brings forth a new set of challenges to overcome. Microfluidics has been a key player in the capture of CTCs and their downstream analysis, with the aim of shedding light into their clinical application in cancer and metastasis. Researchers have taken diverging paths to isolate such cells from blood, ranging from affinity-based isolation targeting surface antigens expressed on CTCs, to label-free isolation taking advantage of the size differences between CTCs and other blood cells. For both major groups, many microfluidic technologies have reported high sensitivity and specificity for capturing CTCs. However, the question remains as to the superiority among these two isolation techniques, specifically to identify different CTC populations. This review highlights the key aspects of affinity and label-free microfluidic CTC technologies, and discusses which of these two would be the highest benefactor for the study of CTCs.
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