Cultured circulating tumor cells and their derived xenografts for personalized oncology.

Recent cancer research has demonstrated the existence of circulating tumor cells (CTCs) in cancer patient's blood. Once identified, CTC biomarkers will be invaluable tools for clinical diagnosis, prognosis and treatment. In this review, we propose ex vivo culture as a rational strategy for large scale amplification of the limited numbers of CTCs from a patient sample, to derive enough CTCs for accurate and reproducible characterization of the biophysical, biochemical, gene expressional and behavioral properties of the harvested cells. Because of tumor cell heterogeneity, it is important to amplify all the CTCs in a blood sample for a comprehensive understanding of their role in cancer metastasis. By analyzing critical steps and technical issues in ex vivo CTC culture, we developed a cost-effective and reproducible protocol directly culturing whole peripheral blood mononuclear cells, relying on an assumed survival advantage in CTCs and CTC-like cells over the normal cells to amplify this specified cluster of cancer cells.

Combined Lung-Kidney Transplantation: An Analysis of the UNOS/OPTN Database.

Poor outcomes after thoracic transplantation with concurrent renal dysfunction are well described: without transplantation or with thoracic-only transplantation, patients face unacceptably high mortality. Outcomes after combined lung-kidney transplantation (LKT) remain largely uninvestigated. The United Network for Organ Sharing/Organ Procurement and Transplantation Network database was queried to identify all LKTs, lung transplantations (LTs), and kidney transplantations (KTs) performed in the United States from 1995 to 2013. Survival was calculated using the Kaplan-Meier method and compared using log-rank tests or Cox regression models. Thirty-one LKTs were performed. Mean recipient age was 45.4 ± 13.5 years; 48.3 per cent were male. Retransplantation for graft failure was the leading indication for LT (n = 13) and the most common renal indication was calcineurin inhibitor nephrotoxicity (n = 11). Mean lung allocation score was 46.6 ± 14.4, mean creatinine was 3.7 ± 2.8 g/dL, and glomerular filtration rate was 23.1 (interquartile range 11.9, 38.3) mL/min/1.7 m(2), and 11 (35.5%) were dialysis dependent. Patient survival after LKT was 92.9 per cent, 71.0 per cent, and 71.0 per cent at one month, six months, and one year, with a median survival of 95.2 months. One- and five-year survival after LKT, 71.0 per cent and 59.9 per cent, were similar to LT (n = 23,913), 81.7 per cent and 51.4 per cent (P = 0.061 and 0.55), and inferior to KT (n = 175,269), 94.9 per cent and 82.8 per cent (P < 0.0001), respectively. Patient survival after LKT was similar to isolated LT, and these results suggest that LKT is a feasible therapeutic option for LT candidates with significant renal dysfunction.

Outcomes of vascular resection in pancreaticoduodenectomy: single-surgeon experience.

Extension of pancreatic adenocarcinoma into adjacent vasculature often necessitates resection of the portal vein (PV) and/or superior mesenteric vein (SMV) during pancreaticoduodenectomy (PD). The vein is reconstructed primarily by end-to-end anastomosis of vein remnants or venoplasty or by use of autologous or synthetic vein grafts. The objective of this study was to review outcomes in patients undergoing PD for pancreatic adenocarcinoma, specifically comparing the short- and long-term outcomes between the patients undergoing vascular resection and those undergoing standard PD. All patients undergoing PD for pancreatic adenocarcinoma by a single surgeon between 2007 and 2012 were reviewed. Of the 61 patients identified, 18 patients underwent vascular resection of the PV (four patients), SMV (10 patients), or both (four patients). The remaining 43 patients had standard PD. Demographic, perioperative, pathological, and long-term outcomes data were collected and both vascular and standard groups were compared. Both groups had similar demographics. The vascular group had significantly longer operative times (529 vs 406 minutes; P < 0.01) with a trend to greater estimated blood loss (0.64 vs 0.53 L; P = 0.06). Pathological analysis showed no difference between the two groups with regard to lymph node status/ratio and rate of R0 resection (94 vs 91%; P = 0.57); however, the size of the tumor was significantly greater in the vascular group (4.2 vs 3 cm; P < 0.01). Short-term outcomes were similar in the vascular group and standard group, respectively, with no difference in pancreatic fistula rate (6 vs 7%; P = 1.0), transfusion rate (44 vs 35%; P = 0.57), and median length of stay (8 vs 7 days; P = 0.10), and there was no 30-day mortality in either group. Based on Kaplan-Meier methods, the median recurrence-free survival was 18 versus 23 months (P = 0.37) in the vascular and standard groups, respectively, and the overall survival was almost identical in both groups, each with a median of 31 months (P = 0.91). In our experience, mesenteric and PV resection during PD was performed safely and without compromise of short- or longer-term outcomes. It can be performed safely and patients have no significant difference in perioperative outcomes or overall survival.

Development of a two-color fluorescence in situ hybridization technique for species-level identification of human-infectious Cryptosporidium spp.

A two-color fluorescence in situ hybridization assay that allows for the simultaneous identification of Cryptosporidium parvum and C. hominis was developed. The assay is a simple, rapid, and cost-effective tool for the detection of the major Cryptosporidium species of concern to public health.

Development of fluorescent in situ hybridization for Cryptosporidium detection reveals zoonotic and anthroponotic transmission of sporadic cryptosporidiosis in Sydney.

Cryptosporidium is the most common non-viral cause of diarrhea worldwide. Of the 5 described species that contribute to the majority of human infections, C. parvum is of major interest due to its zoonotic potential. A species-specific fluorescence in situ hybridisation probe was designed to the variable region in the small subunit of the 18S rRNA of C. parvum and labeled with Cy3. Probe specificity was validated against a panel of 7 other Cryptosporidium spp. before it was applied to 33 human faecal samples positive for cryptosporidiosis which were obtained during the period from 2006-2007. Results were compared to PCR-RFLP targeting the 18S rDNA. FISH results revealed that 19 of the 33 isolates analysed were identified as C. parvum. Correlation of PCR-RFLP and FISH was statistically significant (P<0.05), resulting in a calculated correlation coefficient of 0.994. In this study, species identification by FISH and PCR-RFLP provided preliminary evidence to support both anthroponotic and zoonotic transmission of sporadic cases of cryptosporidiosis in the Sydney basin. In conclusion, FISH using a C. parvum-specific probe provided an alternative tool for accurate identification of zoonotic Cryptosporidium which will be applied in the future to both epidemiological and outbreak investigations.

Massive intracranial immature teratoma with extracranial extension into oral cavity, nose, and neck.

We present a case of congenital massive intracranial teratoma with extracranial extension into oral cavity, nose, and neck diagnosed by antenatal ultrasonography at 25 weeks of gestation. The fetus was delivered by cesarean section because of massive fetal head size and severe maternal pregnancy-induced hypertension. At necropsy the tumor was found to be an immature teratoma, with no recognizable normal brain tissue. An additional finding was hepatomegaly secondary to extensive extramedullary hematopoiesis. Karyotyping of both the fetus and the teratoma revealed a normal female chromosomal composition: 46,XX.

Impact of transfusion guidelines on neonatal transfusions.

BACKGROUND: Preterm infants with birth weights (BW) <1250 gm are given multiple blood transfusions either for replacement of blood loss or for correction of symptomatic anemia of prematurity. OBJECTIVE: To assess the effectiveness of transfusion guidelines in reducing the number of transfusions given to infants with BW <1250 gm. METHODS: This cohort study was conducted at the regional teaching medical center with level III obstetric and neonatal services. Preterm infants with BW <1250 gm and gestational age <32 weeks were admitted to the neonatal intensive care unit during a period of 6 months before and after implementation of transfusion guidelines. The final sample size constituted 39 infants before guidelines (BG) and 41 infants after guidelines (AG). The primary outcome measure was the total number and volume of transfusions given per infant in the first 2 weeks of life and before discharge from the nursery. RESULTS: We observed a significant reduction in the mean number (4.7 to 2.7, p = 0.003) and volume (52 ml to 30 ml, p = 0.0005) of transfusions given per infant in the first 2 weeks of life, as well as a definite trend toward reduction in the total number (10.5 to 8.0, p = 0.08) and volume (156 ml to 119 ml, p = 0.07) of transfusions given before discharge in the BG versus AG, respectively. When all the transfusions given to the 41 infants in the AG group were analyzed for compliance with guidelines, 96% (313 of 328) were observed to be in compliance. CONCLUSION: This study shows that transfusion guidelines are effective in decreasing the number of transfusion given to infants with BW <1250 gm. An indirect benefit of guidelines contributing to a reduced number of transfusions may be a heightened awareness to decrease blood losses.

Systemic hypertension in very low-birth weight infants with bronchopulmonary dysplasia: incidence and risk factors.

The purpose of our study was to determine if systemic hypertension (HTN) occurred among infants with birth weight less than 1250 g (very low-birth-weight [VLBW] infants) in association with Bronchopulmonary dysplasia (BPD). We designed a historical cohort study to review the clinical course and the occurrence of systemic HTN in infants born during the year 1992 with birth weights between 600-1250 g. The overall incidence of HTN was 6.8% (5 of 73) and the incidence in infants with BPD was 12% (5 of 41). The mean age of onset of HTN was 105 days (range 90 to 133 days), and at the time of discharge 3 of 5 (60%) infants remained hypertensive and 3 of 5 (60%) were on supplemental oxygen. All the five hypertension infants (100%) were on supplemental oxygen at 36 weeks of postceptional age compared to 18 of 36 (50%) of nonhypertensive BPD infants. The association between HTN and severe BPD was further denoted by longer hospital stay (145 +/- 37 vs. 94 +/- 28 days, p = 0.004), longer duration of O2 therapy (108 +/- 36 vs. 67 +/- 34 days, p = 0.01), and prolonged use of aminophylline (104 +/- 44 vs. 61 +/- 23 days, p = 0.03), in the hypertensive BPD infants versus nonhypertensive BPD infants, respectively. This study substantiates an increased risk of developing systemic HTN, among VLBW infants with severe BPD.