Assessment of vascular endothelial growth factor in formalin fixed, paraffin embedded colon cancer specimens by means of a well-based reverse phase protein array.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a critical pro-angiogenic factor, found in a number of cancers, and a target of therapy. It is typically assessed by immunohistochemistry (IHC) in clinical research. However, IHC is not a quantitative assay and is rarely reproducible. We compared VEGF levels in colon cancer by IHC and a quantitative immunoassay on proteins isolated from formalin fixed, paraffin embedded tissues. RESULTS: VEGF expression was studied by means of a well-based reverse phase protein array (RPPA) and immunohistochemistry in 69 colon cancer cases, and compared with various clinicopathologic factors. Protein lysates derived from formalin fixed, paraffin embedded tissue contained measurable immunoreactive VEGF molecules. The VEGF expression level of well differentiated colon cancer was significantly higher than those with moderately and poorly differentiated carcinomas by immunohistochemistry (P = 0.04) and well-based RPPA (P = 0.04). VEGF quantification by well-based RPPA also demonstrated an association with nodal metastasis status (P = 0.05). In addition, the normalized VEGF value by well-based RPPA correlated (r = 0.283, P = 0.018). Furthermore, subgroup analysis by histologic type revealed that adenocarcinoma cases showed significant correlation (r = 0.315, P = 0.031) between well-based RPPA and IHC. CONCLUSIONS: The well-based RPPA method is a high throughput and sensitive approach, is an excellent tool for quantification of marker proteins. Notably, this method may be helpful for more objective evaluation of protein expression in cancer patients.

Direct transcriptional regulation of Six6 is controlled by SoxB1 binding to a remote forebrain enhancer.

Six6, a sine oculis homeobox protein, plays a crucial and conserved role in the development of the forebrain and eye. To understand how the expression of Six6 is regulated during embryogenesis, we screened ~250 kb of genomic DNA encompassing the Six6 locus for cis-regulatory elements capable of directing reporter gene expression to sites of Six6 transcription in transgenic mouse embryos. Here, we describe two novel enhancer elements, that are highly conserved in vertebrate species and whose activities recapitulate Six6 expression in the ventral forebrain and eye, respectively. Cross-species comparisons of the Six6 forebrain enhancer sequences revealed highly conserved binding sites matching the consensus for homeodomain and SoxB1 transcription factors. Deletion of either of the binding sites resulted in loss of the forebrain enhancer activity in the ventral forebrain. Moreover, our studies show that members of the SoxB1 family, including Sox2 and Sox3, are expressed in the overlapping region of the ventral forebrain with Six6 and can bind to the Six6 forebrain enhancer. Loss of function of SoxB1 genes in vivo further emphasizes their role in regulating Six6 forebrain enhancer activity. Thus, our data strongly suggest that SoxB1 transcription factors are direct activators of Six6 expression in the ventral forebrain.

Sonography for determining the optic nerve sheath diameter with increasing intracranial pressure in a porcine model.

OBJECTIVES: This study investigated whether it is feasible to use sonography to monitor changes in the optic nerve sheath diameter in a porcine model. METHODS: A fiber-optic intracranial pressure transducer was surgically placed through the frontal sinus directly into the brain parenchyma of adult Yorkshire pigs (n = 5). A second bolt was placed on the contralateral side for intraparenchymal fluid infusion. Optic nerve sheath diameter measurements were acquired by each of 2 ultrasound operators around the leading edge of the nerve, 3 to 5 mm distal from the origin of the optic nerve. To induce a change in diameter, intracranial pressure was manipulated by injecting normal saline into the intraparenchymal infusion catheter located in the symmetric contralateral position as the pressure-monitoring probe. RESULTS: Data from 1 pig were unusable because of a cerebrospinal fluid leak into the sinus and orbital fissure. Saline aliquots of 1 to 10 mL were able to generate intracranial pressures typically starting from 10 to 15 mm Hg and increasing to 75 to 90 mm Hg, which eventually evoked a Cushing response. Fluid injection was controlled to increase pressures by 60 mm Hg over a 15- to 20-minute period. Regression analysis of all animals showed that the optic nerve sheath diameter increased by 0.0034 mm/mm Hg of intracranial pressure; however, this slope ranged from 0.0025 to 0.0046, depending on the animal measured. There was no discernible effect of the ultrasound operator on the slope; however, measurements made by 1 operator were consistently higher than the others by about 8% of the overall diameter range. CONCLUSIONS: These results suggest that the use of the optic nerve sheath diameter to noninvasively confirm acute changes in intracranial pressure over 1 hour is feasible in a porcine model. We recommend that this method be validated in humans using direct intracranial pressure measurement where possible to confirm it as a screening tool for acute and chronically increased diameters secondary to elevated pressure in clinical settings.

Local resection versus radical surgery for parathyroid carcinoma: A National Cancer Database analysis.

INTRODUCTION: Parathyroid carcinoma (PC) is rare and often diagnosed incidentally after local resection (LR) for other indications. Although recommended treatment has traditionally been radical surgery (RS), more recent guidelines suggest that LR alone may be adequate. We sought to further investigate outcomes of RS versus LR for localized PC. MATERIALS AND METHODS: PC patients from 2004 to 2015 with localized disease were identified from the National Cancer Database, then stratified by surgical therapy: LR or RS. Demographic and clinicopathologic data were compared. Cox proportional hazard models were constructed to estimate associations of variables with overall survival (OS). OS was estimated from time of diagnosis using Kaplan-Meier curves. RESULTS: A total of 555 patients were included (LR = 522, RS = 33). The groups were comparable aside from LR patients having higher rates of unknown nodal status (66.9% versus 39.4%; p = 0.003). By multivariable analysis, RS did not have a significant association with OS (hazard ratio (HR) = 0.43, 95% confidence interval (95%CI) = 0.10, 1.83; p = 0.255), nor did positive nodal status (HR = 0.66, 95%CI = 0.09, 5.03; p = 0.692) and unknown nodal status (HR = 1.30, 95%CI = 0.78, 2.17; p = 0.311). There was no difference in OS between the LR and RS groups, with median survival not reached by either group at 10 years (median follow-up = 60.4 months; p = 0.20). CONCLUSIONS: There was no difference in OS between LR and RS for localized PC. RS and nodal status may not impact survival as previously identified, and LR should remain a valid initial surgical approach. Future higher-powered studies are necessary to assess the effects of surgical approaches on morbidity and oncologic outcomes.

Treatment with bone marrow-derived stromal cells accelerates wound healing in diabetic rats.

Bone marrow stem cells participate in tissue repair processes and may have a role in wound healing. Diabetes is characterised by delayed and poor wound healing. We investigated the potential of bone marrow-derived mesenchymal stromal cells (BMSCs) to promote healing of fascial wounds in diabetic rats. After manifestation of streptozotocin (STZ)-induced diabetic state for 5 weeks in male adult Sprague-Dawley rats, healing of fascial wounds was severely compromised. Compromised wound healing in diabetic rats was characterised by excessive polymorphonuclear cell infiltration, lack of granulation tissue formation, deficit of collagen and growth factor [transforming growth factor (TGF-beta), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor PDGF-BB and keratinocyte growth factor (KGF)] expression in the wound tissue and significant decrease in biomechanical strength of wounds. Treatment with BMSC systemically or locally at the wound site improved the wound-breaking strength (WBS) of fascial wounds. The improvement in WBS was associated with an immediate and significant increase in collagen levels (types I-V) in the wound bed. In addition, treatment with BMSCs increased the expression of growth factors critical to proper repair and regeneration of the damaged tissue moderately (TGF-beta, KGF) to markedly (EGF, VEGF, PDGF-BB). These data suggest that cell therapy with BMSCs has the potential to augment healing of the diabetic wounds.

Successful embolization of hepatocelluar carcinoma with yttrium-90 glass microspheres prior to liver transplantation.

We report a case of a patient with end-stage liver disease secondary to hepatitis C, complicated by a large hepatocellular carcinoma. Because of the size of the tumor exceeded the Milan criteria, he was not a candidate for liver transplantation. However, after two treatments with yttrium-90 glass microsphere infusions, the tumor became smaller and the patient's alpha-fetoprotein level dropped to normal range. He was listed for transplantation and subsequently received a deceased donor liver transplant. Two years after his transplantation, he remains tumor free and has normal alpha-fetoprotein levels. This is the first reported case in the literature of using yttrium-90 microspheres as a bridge to liver transplantation in a patient with a large hepatocellular carcinoma. This therapy should be considered in patients with cirrhosis and large hepatocellular carcinomas exceeding current size criterion, who would otherwise be good candidates for transplantation.

Safe introduction of a new surgical technique: remote telementoring for posterior retroperitoneoscopic adrenalectomy.

BACKGROUND: Posterior retroperitoneoscopic adrenalectomy (PRA) is a safe and effective approach to adrenalectomy, offering less pain and faster recovery than open or laparoscopic surgery. Although the popularity of PRA is increasing, few surgical centres have extensive experience with the procedure. The ideal approach to achieve proficiency with any new technique involves on-site observation of an experienced surgeon-mentor, followed by mentored hands-on experience of the surgeon-learner. However, it is not always feasible for a surgeon-mentor to offer on-site supervision to the surgeon-learner in his or her home institution. Advances in Internet applications have made remote telementoring a viable alternative to on-site mentoring in selected situations. METHODS: We describe our experience in safely introducing PRA to Melbourne, Australia, where no highly experienced surgeon-mentors were available. A surgeon with experience of 12 PRA procedures attended from interstate, along with live telementoring via Skype video link by an overseas surgeon who had performed more than 200 PRA procedures, to mentor the surgeon-learner performing her first three cases. RESULTS: The operating surgeon's first three PRA procedures proceeded uneventfully, with no complications, relatively short operative times and one-night hospital stays for all three patients. Twenty-two more have been performed since, without complications. CONCLUSION: Remote telementoring is a safe and feasible way to assist surgeons in safely introducing new techniques. This strategy is particularly applicable in centres where no surgeon-mentor is locally available. It is important that the surgeon-learner has the skills and experience to complete the procedure using alternative techniques in the case of complications or technical failure.

The role of minimally invasive surgery and outcomes in colorectal cancer.

For some time now, there has been significant interest in understanding and defining the role of minimally invasive surgery in colorectal cancer. Laparoscopic surgery has been shown to have similar or better outcomes compared with open surgery. Recently, prospective randomized trials have demonstrated oncologic outcomes of laparoscopic colon surgery equivalent to those for open surgery. However, the technical challenges of performing laparoscopic resection of rectal cancers and the uncertainty of the oncologic quality of the surgical resection have hindered the growth of minimally invasive rectal surgery. Robotic rectal surgery has recently emerged as an attractive alternative to laparoscopic surgery because it allows for superior visualization within a narrow pelvic field and more precise dissection. Studies of robotic rectal resection have suggested similar or potentially improved short-term oncologic outcomes when compared with laparoscopic rectal resection. Ongoing randomized studies will provide additional insight into the role of laparoscopic and minimally invasive robotic surgery for rectal cancer.

Rare Diagnosis of a Common Supraventricular Tachycardia.

This case describes diagnostic maneuvers that will lead to an accurate diagnosis of a given A on V supraventricular tachycardia. The list of maneuvers described is not exhaustive but includes techniques to help diagnose a concealed NV/NF pathway mediated tachycardia that is in the differential diagnosis of a failed atrioventricular nodal re-entrant tachycardia ablation. The practicing electrophysiologist must be prepared to perform these maneuvers in order to ensure the safest and most focused ablation procedures.

Atrial Flutter Late After Remote Tetralogy of Fallot Repair.

This case illustrates an atypical presentation of typical atrial flutter, demonstrating the importance of always considering cavo-tricuspid isthmus dependent atrial flutter as the potential culprit.

Immunohistochemical identification of primary peritoneal serous cystadenocarcinoma mimicking advanced colorectal carcinoma: a case report.

Primary peritoneal cystadenocarcinoma is a rare tumor of similar histogenic origin as primary ovarian carcinoma. We present a case of primary peritoneal serous cystadenocarcinoma mimicking advanced colorectal cancer in a 68 yr-old African American female. Radiology, endoscopy and cytology yielded only inconclusive findings. Immunohistochemical analysis of percutaneously obtained ascitic fluid provided a correct diagnosis of primary peritoneal cystadenocarcinoma. The discovery of serous ascites at the time of laparotomy confirmed a diagnosis of primary peritoneal serous cystadenocarcinoma. Final surgical pathology reconfirmed the diagnosis of primary peritoneal cystadenocarcinoma. This case demonstrates the utility of immunohistochemistry for accurately diagnosing patients with inconclusive findings in the setting of peritoneal carcinomatosis and primary peritoneal cystadenocarcinoma.

Physiologic subperiosteal new bone formation: prevalence, distribution, and thickness in neonates and infants.

OBJECTIVE: The objective of our study was to determine the prevalence, distribution, and thickness of physiologic subperiosteal new bone formation in neonates and infants. MATERIALS AND METHODS: High-detail postmortem skeletal radiologic surveys of 101 neonates and infants who had died from sudden infant death syndrome were reviewed. The average age at the time of death was 2.6 months (range, 2 weeks-8 months 2 weeks). The location, distribution, and thickness of subperiosteal new bone formation of the long bones were noted and measured with an ocular magnification system. RESULTS: subperiosteal new bone formation was identified in 35 infants (35%), all of whom were between 1 and 4 months of age. The prevalence of subperiosteal new bone formation involving one or more bones was 37% for ages 1-2 months, 55% for ages 2-3 months, and 35% for ages 3-4 months. subperiosteal new bone formation involved the tibia in 29 infants (bilateral involvement in 19) and the femurs in 14 infants (bilateral involvement in 11). The humerus was a site of subperiosteal new bone formation in 12 infants; the ulna, in seven infants; and the radius, in two infants. The thickness of the subperiosteal new bone formation was never more than 1.8 mm, and the average thickness for the various long bones ranged from 0.7 to 0.9 mm. CONCLUSION: Our results suggest that subperiosteal new bone formation is a common finding in infants 1-4 months of age. It is potentially an abnormal finding in neonates (CI = 0.0, 0.007) and in infants older than 4 months of age (CI = 0.0, 0.003). A thickness of 2 mm or more in subperiosteal new bone formation is likely to be abnormal and should prompt further diagnostic evaluation.