The cancer-screening dilemma

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Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced in vitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced in vitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves (p < 0.05). On the other hand, no differences were found in the development of bovine embryos in vitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs.     

Insulin and blood pressure in obesity

To assess factors in overweight persons that account for a tendency toward hypertension, 33 very obese women, 26 to 77 years of age, were studied. Blood pressures in these 33 women varied from low normal to mildly hypertensive. None of them had taken medication for high blood pressure, and none had diabetes mellitus. The effect of independent variables--age, body mass index (weight/height2), fasting serum glucose levels, fasting serum insulin levels, and 24-hour urinary sodium excretion--on systolic and diastolic blood pressure was assessed. There was no correlation between sodium excretion and blood pressure. Age did not correlate with diastolic blood pressure but did correlate with systolic blood pressure when body mass index, serum glucose level, and insulin level were controlled. Diastolic blood pressure correlated with body mass index and serum glucose level, but only the latter remained significant when all independent variables were considered together. Both systolic and diastolic blood pressure were found to be significantly related to fasting serum insulin level (r = 0.47, p = 0.005 and r = 0.68, p less than 0.001) even when age, weight, and serum glucose level were controlled (r = 0.41, p = 0.025 and r = 0.62, p less than 0.001 respectively). The relation between serum insulin and blood pressure was more pronounced in those women with a family history of hypertension. These data indicate that insulin may play a major role in the regulation of blood pressure in obesity and that the previously accepted relation of weight to blood pressure may depend on blood levels of insulin.     

The use of intraoperative ultrasound in laparoscopic adrenal surgery

Background: Laparoscopic adrenalectomy has been shown to be a safe and effective therapy for benign adrenal lesions. We review our experience with this procedure, including the use of laparoscopic ultrasound. Methods: We retrospectively reviewed our experience with 36 patients who underwent resection of 42 adrenal glands. Data gathered included preoperative evaluation and diagnosis, operative time, blood loss, complications, and follow-up status. Laparoscopic ultrasound was used to guide dissection and characterize a variety of adrenal lesions. Results: Thirty-five of 36 patients underwent successful laparoscopic adrenalectomy. There was one conversion to the open procedure in a patient with bilateral adrenal metastases from an endometrial cancer. For the bilateral laparoscopic procedure, the operative time averaged 262 mins, blood loss was 160 cc, and hospital stay was 3.0 days. For unilateral cases, operative time averaged 193 min, blood loss was 108 cc, and hospitalization was 1.1 days. Six patients experienced perioperative complications, most of which were minor and transient. Laparoscopic ultrasound was useful to define anatomy and to identify the adrenal vein, especially on the left side. Conclusions: Laparoscopic adrenalectomy is the procedure of choice for benign adrenal disease. Laparoscopic ultrasound is useful to localize and aid in the dissection of the left adrenal vein. Received: 24 December 1998/Accepted: 12 February 1999     

Arterial oxygenation during one-lung ventilation: combined versus general anesthesia

The optimal anesthetic management of patients undergoing thoracotomy for pulmonary resection has not been definitely determined. We evaluated whether general i.v. anesthesia (propofol-fentanyl) provides superior PaO2 during one-lung ventilation (OLV) compared with thoracic epidural anesthesia (TEA) with supplemental local and general anesthetics. We studied 60 patients who had prolonged periods of OLV for elective thoracic surgery for lung cancer and who were prospectively randomized into two groups. In 30 patients (GA group), fentanyl/propofol/rocuronium anesthesia was used. Another 30 patients (TEA group) were anesthetized with propofol/rocuronium/epidural thoracic bupivacaine 0.5%. A double-lumen endotracheal tube was inserted, and mechanical ventilation with 100% oxygen was used during the entire study. Arterial and venous blood gases were recorded before surgery in a lateral position with two-lung ventilation, 15 and 30 min after OLV (OLV + 15 and OLV + 30, respectively) in all patients. We measured PaO2, venous central oxygen tension, arterial and central venous oxygen saturation, venous admixture percentage (Qs/Qt%), and arterial and central venous oxygen content. The mean values for PaO2 during OLV in the GA group after 15 min (175 mm Hg) and 30 min (182 mm Hg) were significantly (P < 0.05) higher compared with the TEA group (120 and 118 mm Hg, respectively). Furthermore, Qs/Qt% was significantly (P < 0.05) increased in the TEA group during OLV. There were no other significant differences. We conclude that using the TEA regimen is associated with a lower PaO2 and a larger intrapulmonary shunt during OLV than with total i.v. anesthesia alone. IMPLICATIONS: Sixty patients undergoing elective lung surgery during a prolonged period of intraoperative one-lung ventilation were studied and randomized to receive general i.v. anesthesia or general i.v. anesthesia combined with thoracic epidural anesthesia. The arterial oxygenation in the first group was better than that in the second group during one-lung ventilation.     

Continuous and intermittent running to exhaustion at maximal lactate steady state: Neuromuscular,biochemical and endocrinal responses

ObjectiveThe aim of the present study was to characterize the neuromuscular, biochemical, and endocrinal responses from a running to exhaustion mode at the maximal lactate steady state intensity during continuous and intermittent protocols.DesignPre-post test measures.MethodsTwelve athletes performed an incremental treadmill test, several constant speed tests to determine the maximal lactate steady state at continuous and intermittent (5:1 ratio) models and two randomized tests until exhaustion at such intensities. Knee extension torque and blood sampling were collected before and immediately after the time to exhaustion tests.ResultsThe results showed a significant decrement (～15%) in torque production after time to exhaustion tests for both exercise models. In addition to neuromuscular impairment, an acute increase of 65% and 38% was observed creatine kinase, during continuous and intermittent running, respectively. Regarding hormonal responses when compared to baseline measurements, cortisol increased by 132% and 121% in the continuous and intermittent protocols, respectively. No correlation was found between biochemical, endocrinal and the neuromuscular variables.ConclusionsThe present findings showed that running until exhaustion performed at maximal lactate steady state, significantly impaired muscle strength and increased hormonal and muscle damage markers in two different protocols (i.e. continuous and intermittent) amongst trained runners.