The MRI scan of his head revealed a small mass (5 mm) in the pitu

The MRI scan of his head revealed a small mass (5 mm) in the pituitary gland. The CT scan revealed

Opaganib purchase two pancreatic tumors; a 3.8 cm mass in the head of the pancreas (Figure 1 left) and a 1.2 cm mass in the body of the pancreas (Figure 1 right, arrowheads). His surgical treatment included resection of the head of the pancreas (Whipple’s procedure) and enucleation of the tumor from the body of the pancreas. Within 1 week of surgery, his diarrhea had resolved and his serum potassium had returned to normal. Histology revealed neuroendocrine tumors of uncertain malignant potential (Figure 2, above) with positive immunohistochemical staining for chromogranin A and vasoactive intestinal peptide (VIP, Figure 2, below). Selumetinib research buy The neoplasms were also focally positive for glucagon but negative for proinsulin, gastrin, serotonin and somatostatin. His family history was helpful as his father had complicated peptic ulcer disease caused by a pancreatic gastrinoma and was treated with a subtotal gastrectomy. MENI is an uncommon disease with a prevalence of approximately 1:30,000 people. It is caused by mutations in the MENI gene that encodes a protein called menin. The most common clinical manifestation is hyperparathyroidism that occurs in approximately 90% of patients. Most patients also develop neoplasms in

the pancreas that may be non-functional or may result in the secretion of hormones such as gastrin, insulin, glucagon, somatostatin and VIP. VIPomas are extremely rare with an estimated annual incidence of 1:10 million people. With immunocytochemistry, some VIPomas can have positive staining with other hormones such as pancreatic polypeptide, glucagon and somatostatin. For patients without metastases, medchemexpress the treatment of choice is surgical excision of the neoplasms. This usually results in improvement or resolution of diarrhea. Contributed by “
“Sayin SI, Wahlstrom A, Felin J, Jantti S, Marschall HU, Bamberg

K, et al. Gut microbiota regulates bile acid metabolism by reducing the levels of tauro-beta-muricholic acid, a naturally occurring FXR antagonist. Cell Metab 2013;17:225-235. (Reprinted with permission). Bile acids are synthesized from cholesterol in the liver and further metabolized by the gut microbiota into secondary bile acids. Bile acid synthesis is under negative feedback control through activation of the nuclear receptor farnesoid X receptor (FXR) in the ileum and liver. Here we profiled the bile acid composition throughout the enterohepatic system in germfree (GF) and conventionally raised (CONV-R) mice. We confirmed a dramatic reduction in muricholic acid, but not cholic acid, levels in CONV-R mice.

Conclusion: Transplantation studies indicate that the state of th

Conclusion: Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure. (HEPATOLOGY 2011) Expansion and altered composition of the extracellular matrix as a result of collagen deposition is a common response to injury and plays a major role in chronic heart, liver, and kidney

failure. Understanding the extent to which reversal of this process can lead to functional organ recovery is a critical issue, as numerous interventions have been proposed to improve fibrosis and presumably reverse organ failure.1-4 The unique capacity of hepatic parenchymal cells to undergo extensive proliferation in response to injury makes the liver Small molecule library an ideal organ to study cellular regeneration in acquired chronic disease. In the liver, expansion of the extracellular matrix with capillarization of the sinusoidal endothelium and loss of fenestrae results in cirrhosis with production learn more of regenerative hepatic nodules, portal hypertension, loss of hepatocytes, and liver failure.5 Loss of significant hepatocyte mass does not routinely produce hepatic failure, because the liver is capable

of normal function with less than half its normal complement of hepatocytes.6, 7 Thus, the cause of organ failure in cirrhosis is not well understood. Impaired hepatic function results from intrinsic damage to the native liver cells and from the abnormal

microenvironment in which they reside.8-14 Because collagen deposition and vascular changes in cirrhosis can be extensive before there is functional hepatic decompensation, it is not clear to what extent each plays a role or at what point these factors tip parenchymal cell function toward organ failure. Mito et al.15 attempted to address the role of the microenvironment in hepatic failure by transplanting 上海皓元 hepatocytes from the livers of patients with cirrhosis back into their own spleens to reverse decompensated liver disease. If it is possible to recover the function of parenchymal cells from a cirrhotic liver by changing the microenvironment, it may be possible to restore hepatic function in the cirrhotic liver by reversing hepatic structural abnormalities, and individual cells derived from some cirrhotic livers might prove to be useful as an untapped source of transplantable cells for the treatment of patients with liver-based metabolic disorders, where the liver microenvironment is intact. Here, we demonstrate that primary cells derived from cirrhotic livers with decompensated function exhibit severe alterations in gene expression and defects in proliferative capacity and function directly after isolation, but engraft normally in a noncirrhotic microenvironment.

Conclusion: Transplantation studies indicate that the state of th

Conclusion: Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure. (HEPATOLOGY 2011) Expansion and altered composition of the extracellular matrix as a result of collagen deposition is a common response to injury and plays a major role in chronic heart, liver, and kidney

failure. Understanding the extent to which reversal of this process can lead to functional organ recovery is a critical issue, as numerous interventions have been proposed to improve fibrosis and presumably reverse organ failure.1-4 The unique capacity of hepatic parenchymal cells to undergo extensive proliferation in response to injury makes the liver Small molecule library high throughput an ideal organ to study cellular regeneration in acquired chronic disease. In the liver, expansion of the extracellular matrix with capillarization of the sinusoidal endothelium and loss of fenestrae results in cirrhosis with production TGF-beta inhibitor of regenerative hepatic nodules, portal hypertension, loss of hepatocytes, and liver failure.5 Loss of significant hepatocyte mass does not routinely produce hepatic failure, because the liver is capable

of normal function with less than half its normal complement of hepatocytes.6, 7 Thus, the cause of organ failure in cirrhosis is not well understood. Impaired hepatic function results from intrinsic damage to the native liver cells and from the abnormal

microenvironment in which they reside.8-14 Because collagen deposition and vascular changes in cirrhosis can be extensive before there is functional hepatic decompensation, it is not clear to what extent each plays a role or at what point these factors tip parenchymal cell function toward organ failure. Mito et al.15 attempted to address the role of the microenvironment in hepatic failure by transplanting MCE hepatocytes from the livers of patients with cirrhosis back into their own spleens to reverse decompensated liver disease. If it is possible to recover the function of parenchymal cells from a cirrhotic liver by changing the microenvironment, it may be possible to restore hepatic function in the cirrhotic liver by reversing hepatic structural abnormalities, and individual cells derived from some cirrhotic livers might prove to be useful as an untapped source of transplantable cells for the treatment of patients with liver-based metabolic disorders, where the liver microenvironment is intact. Here, we demonstrate that primary cells derived from cirrhotic livers with decompensated function exhibit severe alterations in gene expression and defects in proliferative capacity and function directly after isolation, but engraft normally in a noncirrhotic microenvironment.

Conclusion: Transplantation studies indicate that the state of th

Conclusion: Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure. (HEPATOLOGY 2011) Expansion and altered composition of the extracellular matrix as a result of collagen deposition is a common response to injury and plays a major role in chronic heart, liver, and kidney

failure. Understanding the extent to which reversal of this process can lead to functional organ recovery is a critical issue, as numerous interventions have been proposed to improve fibrosis and presumably reverse organ failure.1-4 The unique capacity of hepatic parenchymal cells to undergo extensive proliferation in response to injury makes the liver check details an ideal organ to study cellular regeneration in acquired chronic disease. In the liver, expansion of the extracellular matrix with capillarization of the sinusoidal endothelium and loss of fenestrae results in cirrhosis with production selleck chemicals of regenerative hepatic nodules, portal hypertension, loss of hepatocytes, and liver failure.5 Loss of significant hepatocyte mass does not routinely produce hepatic failure, because the liver is capable

of normal function with less than half its normal complement of hepatocytes.6, 7 Thus, the cause of organ failure in cirrhosis is not well understood. Impaired hepatic function results from intrinsic damage to the native liver cells and from the abnormal

microenvironment in which they reside.8-14 Because collagen deposition and vascular changes in cirrhosis can be extensive before there is functional hepatic decompensation, it is not clear to what extent each plays a role or at what point these factors tip parenchymal cell function toward organ failure. Mito et al.15 attempted to address the role of the microenvironment in hepatic failure by transplanting medchemexpress hepatocytes from the livers of patients with cirrhosis back into their own spleens to reverse decompensated liver disease. If it is possible to recover the function of parenchymal cells from a cirrhotic liver by changing the microenvironment, it may be possible to restore hepatic function in the cirrhotic liver by reversing hepatic structural abnormalities, and individual cells derived from some cirrhotic livers might prove to be useful as an untapped source of transplantable cells for the treatment of patients with liver-based metabolic disorders, where the liver microenvironment is intact. Here, we demonstrate that primary cells derived from cirrhotic livers with decompensated function exhibit severe alterations in gene expression and defects in proliferative capacity and function directly after isolation, but engraft normally in a noncirrhotic microenvironment.

Upon the delivery of intracellular model antigens,

Upon the delivery of intracellular model antigens, NVP-AUY922 ic50 hepatocyte-targeted IDLVs induced transgene-specific regulatory T cells that contributed to the observed immune tolerance. Deep sequencing of IDLV-transduced livers showed only rare genomic

integrations that had no preference for gene coding regions and occurred mostly by a mechanism inconsistent with residual integrase activity. Conclusion: IDLVs provide an attractive platform for the tolerogenic expression of intracellular or secreted proteins in the liver with a substantially reduced risk of insertional mutagenesis. (HEPATOLOGY 2011;) “
“Obesity is associated with an aggressive course in chronic viral hepatitis; however, its impact in the development of clinical decompensation (CD) in patients with established cirrhosis is uncertain. We evaluated the role of obesity, in relationship to other recognized predictors, in the development of CD in patients with compensated cirrhosis. The study population, a subset of patients included in a randomized trial of beta-blockers in the prevention of varices in whom data on body mass index (BMI) was available, consisted of 161 patients with compensated cirrhosis. Laboratory tests and portal pressure (assessed by the hepatic venous pressure

gradient or HVPG) were assessed on inclusion. Patients were followed until CD (ascites, hepatic encephalopathy, Selleckchem LDE225 or variceal hemorrhage), or until September 2002. Altogether, 29% had a normal BMI, 40% were overweight, and 30% were obese. In a median follow-up of 59 months, CD occurred in 48/161 (30%) patients with an increasingly higher rate according to BMI group (15% in those with normal BMI; 31% in overweight; 43% in obese patients, P = 0.011). The actuarial probability of developing CD 上海皓元 was significantly higher in the abnormal BMI groups (P = 0.022). In a multivariate model that included parameters previously identified as being predictive of CD (HVPG, albumin,

Mayo endstage liver disease score), etiology, and treatment group, BMI (hazard ration 1.06; 95% confidence interval 1.01-1.12), P = 0.02] was an independent predictor of decompensation, together with HVPG and albumin. Conclusion: Obesity has a deleterious effect on the natural history of compensated cirrhosis of all etiologies, independent of portal pressure and liver function. Weight reduction may be a valuable therapeutic measure in this patient population. (HEPATOLOGY 2011;) The natural history of chronic liver diseases is characterized by the progression of fibrosis and nodule formation leading to the development of cirrhosis. Once cirrhosis is established, patients progress from a frequently asymptomatic compensated stage to a decompensated stage, marked by the development of clinical complications of portal hypertension and liver failure.

Upon the delivery of intracellular model antigens,

Upon the delivery of intracellular model antigens, GSK126 concentration hepatocyte-targeted IDLVs induced transgene-specific regulatory T cells that contributed to the observed immune tolerance. Deep sequencing of IDLV-transduced livers showed only rare genomic

integrations that had no preference for gene coding regions and occurred mostly by a mechanism inconsistent with residual integrase activity. Conclusion: IDLVs provide an attractive platform for the tolerogenic expression of intracellular or secreted proteins in the liver with a substantially reduced risk of insertional mutagenesis. (HEPATOLOGY 2011;) “
“Obesity is associated with an aggressive course in chronic viral hepatitis; however, its impact in the development of clinical decompensation (CD) in patients with established cirrhosis is uncertain. We evaluated the role of obesity, in relationship to other recognized predictors, in the development of CD in patients with compensated cirrhosis. The study population, a subset of patients included in a randomized trial of beta-blockers in the prevention of varices in whom data on body mass index (BMI) was available, consisted of 161 patients with compensated cirrhosis. Laboratory tests and portal pressure (assessed by the hepatic venous pressure

gradient or HVPG) were assessed on inclusion. Patients were followed until CD (ascites, hepatic encephalopathy, Selleckchem Erlotinib or variceal hemorrhage), or until September 2002. Altogether, 29% had a normal BMI, 40% were overweight, and 30% were obese. In a median follow-up of 59 months, CD occurred in 48/161 (30%) patients with an increasingly higher rate according to BMI group (15% in those with normal BMI; 31% in overweight; 43% in obese patients, P = 0.011). The actuarial probability of developing CD medchemexpress was significantly higher in the abnormal BMI groups (P = 0.022). In a multivariate model that included parameters previously identified as being predictive of CD (HVPG, albumin,

Mayo endstage liver disease score), etiology, and treatment group, BMI (hazard ration 1.06; 95% confidence interval 1.01-1.12), P = 0.02] was an independent predictor of decompensation, together with HVPG and albumin. Conclusion: Obesity has a deleterious effect on the natural history of compensated cirrhosis of all etiologies, independent of portal pressure and liver function. Weight reduction may be a valuable therapeutic measure in this patient population. (HEPATOLOGY 2011;) The natural history of chronic liver diseases is characterized by the progression of fibrosis and nodule formation leading to the development of cirrhosis. Once cirrhosis is established, patients progress from a frequently asymptomatic compensated stage to a decompensated stage, marked by the development of clinical complications of portal hypertension and liver failure.

A modified TGA DWI protocols for detecting TGA lesions are useful

A modified TGA DWI protocols for detecting TGA lesions are useful in large-scale clinical practice for confirming the diagnosis of TGA patients with clinical findings. “
“We describe a novel technique for cerebral embolic device placement with inadvertent entrapment and subsequent rescue in the endovascular treatment of innominate artery stenosis. A 62-year-old female presented with symptomatic right-sided subclavian steal syndrome. Single-site access for revascularization of critical innominate artery stenosis with simultaneous cerebral Selleck Y 27632 embolic protection performed for this diagnosis has not been previously reported. Initial nontarget self-expanding

stent deployment within the right subclavian artery resulted in entrapment of the embolic protection device. The device was retrieved through snare fixation and resheathing within a 6-French guide catheter navigated through common femoral artery access. Innominate artery balloon-mounted stent angioplasty was performed preceded by the embolic device retrieval, with complete resolution of symptoms. Endovascular STAT inhibitor distal protection device placement for prevention of cerebral atherothromboembolism during innominate artery stent angioplasty

is not without risk and utilization needs to be carefully considered. “
“Cardiac Echoscan is the simplified transthoracic echocardiogram focused on the main source of emboli detection in the acute stroke diagnosis (Stroke Echoscan). We describe the clinical impact related to the Stroke Echoscan protocol in our Center. Acute stroke patients who underwent the Stroke Echoscan by a trained stroke neurologist were included (Echoscan MCE公司 group). All examinations were reviewed by cardiologists. The main embolic stroke etiologies

were: ventricular akinesia (VA), severe aortic atheroma (AA) plaque and cardiac shunt (SHUNT). The rate of the embolic stroke etiologies and the median length of stay (LOS) were compared with a cohort of patients studied by cardiologist (Echo group). Eighty acute stroke patients were included. The sensitivity (S) and specificity (E) were: VA (S 98.6%, E 66.7%, k = .7), AA (S 93.3%, E 96.9%, k = .88) and SHUNT (S 100%, E 100%, k = 1), respectively. The rate of AA diagnosis was significantly higher in Echoscan group (18.8% vs. 8.9%; P = .05). Echoscan protocol significantly reduced the LOS: 6 days (IQR 3-10) versus Echo group 9 days (IQR 6-13; P < .001). The Echoscan protocol was an accurate quick test, which reduced the length of stay and increased the percentage of severe AA plaque diagnosis. "
“To report a novel endovascular coiling technique for ligation of aneurysms presenting with cranial neuropathy. We describe three patients (all female, median age: 57) presenting with unruptured, mass effect producing, aneurysms. All three were treated with coiling of the aneurysm inflow zone without deploying coils in the dome or fundus.

A modified TGA DWI protocols for detecting TGA lesions are useful

A modified TGA DWI protocols for detecting TGA lesions are useful in large-scale clinical practice for confirming the diagnosis of TGA patients with clinical findings. “
“We describe a novel technique for cerebral embolic device placement with inadvertent entrapment and subsequent rescue in the endovascular treatment of innominate artery stenosis. A 62-year-old female presented with symptomatic right-sided subclavian steal syndrome. Single-site access for revascularization of critical innominate artery stenosis with simultaneous cerebral this website embolic protection performed for this diagnosis has not been previously reported. Initial nontarget self-expanding

stent deployment within the right subclavian artery resulted in entrapment of the embolic protection device. The device was retrieved through snare fixation and resheathing within a 6-French guide catheter navigated through common femoral artery access. Innominate artery balloon-mounted stent angioplasty was performed preceded by the embolic device retrieval, with complete resolution of symptoms. Endovascular Ulixertinib research buy distal protection device placement for prevention of cerebral atherothromboembolism during innominate artery stent angioplasty

is not without risk and utilization needs to be carefully considered. “
“Cardiac Echoscan is the simplified transthoracic echocardiogram focused on the main source of emboli detection in the acute stroke diagnosis (Stroke Echoscan). We describe the clinical impact related to the Stroke Echoscan protocol in our Center. Acute stroke patients who underwent the Stroke Echoscan by a trained stroke neurologist were included (Echoscan 上海皓元医药股份有限公司 group). All examinations were reviewed by cardiologists. The main embolic stroke etiologies

were: ventricular akinesia (VA), severe aortic atheroma (AA) plaque and cardiac shunt (SHUNT). The rate of the embolic stroke etiologies and the median length of stay (LOS) were compared with a cohort of patients studied by cardiologist (Echo group). Eighty acute stroke patients were included. The sensitivity (S) and specificity (E) were: VA (S 98.6%, E 66.7%, k = .7), AA (S 93.3%, E 96.9%, k = .88) and SHUNT (S 100%, E 100%, k = 1), respectively. The rate of AA diagnosis was significantly higher in Echoscan group (18.8% vs. 8.9%; P = .05). Echoscan protocol significantly reduced the LOS: 6 days (IQR 3-10) versus Echo group 9 days (IQR 6-13; P < .001). The Echoscan protocol was an accurate quick test, which reduced the length of stay and increased the percentage of severe AA plaque diagnosis. "
“To report a novel endovascular coiling technique for ligation of aneurysms presenting with cranial neuropathy. We describe three patients (all female, median age: 57) presenting with unruptured, mass effect producing, aneurysms. All three were treated with coiling of the aneurysm inflow zone without deploying coils in the dome or fundus.

The design and primary outcomes of the VIRAHEP-C trial have been

The design and primary outcomes of the VIRAHEP-C trial have been reported elsewhere.8 Adults who were treatment-naïve, infected with genotype 1, had detectable HCV RNA, and had histological evidence of chronic HCV were eligible to participate. Patients were classified by race as either African American or Caucasian, and by ethnicity as either Hispanic or non-Hispanic, based on self-report. All participants were required to have

been born in the United States. From eight clinical centers across the United States, 401 patients were enrolled and began therapy between July 2002 and December 2003. For the present study, serum samples were acquired from a subset of 272 patients from the total VIRAHEP-C cohort, comprising 157 responders Selleckchem Idasanutlin (104 CA, 53 AA) and 115 nonresponders (34 CA, 81 AA). All specimens analyzed were obtained under Institutional Review Board–approved protocols for which participants

provided written informed consent, including consent for genetic testing. Patients received peginterferon alfa-2a (Pegasys; Roche Pharmaceuticals, Nutley, NJ) 180 μg/week and ribavirin (Copegus; Roche Pharmaceuticals) 1,000-1,200 mg/day for at least 24 weeks. Patients who became HCV RNA–negative by week 24 continued treatment for a total of 48 weeks, whereas patients who remained HCV RNA–positive stopped treatment and were considered nonresponders. The primary endpoint of the trial was SVR, defined as the absence of detectable HCV RNA for at least 24 weeks after stopping therapy. HCV RNA testing was performed at a central laboratory (SeraCare BioServices, Gaithersburg, MD) FK228 using the Cobas Amplicor Assay (sensitivity, 50 IU/mL; Roche Molecular Diagnostics, Alameda, CA). Selected samples were tested for HCV RNA levels using the Cobas Amplicor Monitor Assay and for HCV RNA genotype using 上海皓元医药股份有限公司 the Versant HCV Genotype Assay (Bayer, Tarrytown, NY). All patients had undergone liver biopsy within 18 months of screening, and

the biopsy specimens were assessed by a blinded central pathologist. All biopsies were assessed for severity of hepatitis C by grading the inflammation and staging the fibrosis using Ishak’s modified histological activity index. IP-10 was measured in serum samples collected at baseline, prior to initiation of treatment, using the commercially available Quantikine human CXCL10/IP-10 immunoassay (R&D Systems). All samples were diluted 1:2 and analyzed in duplicate. The linear dynamic range of the IP-10 measurement in this assay was 8-500 pg/mL, with a detection limit at 7.8 pg/mL. Samples with IP-10 concentration above 1,000 pg/mL were diluted 1:5 and reanalyzed. The IL28B polymorphic marker rs12979860 was analyzed using the ABI TaqMan allelic discrimination kit and the ABI7900HT Sequence Detection System (Applied Biosystems) as described by Thomas and colleagues.

The design and primary outcomes of the VIRAHEP-C trial have been

The design and primary outcomes of the VIRAHEP-C trial have been reported elsewhere.8 Adults who were treatment-naïve, infected with genotype 1, had detectable HCV RNA, and had histological evidence of chronic HCV were eligible to participate. Patients were classified by race as either African American or Caucasian, and by ethnicity as either Hispanic or non-Hispanic, based on self-report. All participants were required to have

been born in the United States. From eight clinical centers across the United States, 401 patients were enrolled and began therapy between July 2002 and December 2003. For the present study, serum samples were acquired from a subset of 272 patients from the total VIRAHEP-C cohort, comprising 157 responders www.selleckchem.com/products/Vorinostat-saha.html (104 CA, 53 AA) and 115 nonresponders (34 CA, 81 AA). All specimens analyzed were obtained under Institutional Review Board–approved protocols for which participants

provided written informed consent, including consent for genetic testing. Patients received peginterferon alfa-2a (Pegasys; Roche Pharmaceuticals, Nutley, NJ) 180 μg/week and ribavirin (Copegus; Roche Pharmaceuticals) 1,000-1,200 mg/day for at least 24 weeks. Patients who became HCV RNA–negative by week 24 continued treatment for a total of 48 weeks, whereas patients who remained HCV RNA–positive stopped treatment and were considered nonresponders. The primary endpoint of the trial was SVR, defined as the absence of detectable HCV RNA for at least 24 weeks after stopping therapy. HCV RNA testing was performed at a central laboratory (SeraCare BioServices, Gaithersburg, MD) selleck screening library using the Cobas Amplicor Assay (sensitivity, 50 IU/mL; Roche Molecular Diagnostics, Alameda, CA). Selected samples were tested for HCV RNA levels using the Cobas Amplicor Monitor Assay and for HCV RNA genotype using MCE公司 the Versant HCV Genotype Assay (Bayer, Tarrytown, NY). All patients had undergone liver biopsy within 18 months of screening, and

the biopsy specimens were assessed by a blinded central pathologist. All biopsies were assessed for severity of hepatitis C by grading the inflammation and staging the fibrosis using Ishak’s modified histological activity index. IP-10 was measured in serum samples collected at baseline, prior to initiation of treatment, using the commercially available Quantikine human CXCL10/IP-10 immunoassay (R&D Systems). All samples were diluted 1:2 and analyzed in duplicate. The linear dynamic range of the IP-10 measurement in this assay was 8-500 pg/mL, with a detection limit at 7.8 pg/mL. Samples with IP-10 concentration above 1,000 pg/mL were diluted 1:5 and reanalyzed. The IL28B polymorphic marker rs12979860 was analyzed using the ABI TaqMan allelic discrimination kit and the ABI7900HT Sequence Detection System (Applied Biosystems) as described by Thomas and colleagues.