Detection by denaturant gradient gel electrophoresis of tumor-specific mutations in biopsies and relative bronchoalveolar lavage fluid from resectable non-small cell lung cancer.

A PCR-denaturant gradient gel electrophoresis (DGGE) method was developed for the detection of p53 and K-ras mutations in primary operable tumors and paired BAL samples of non-small cell lung cancer. Among 36 patients, 9 showed p53 exon V mutations in biopsies and in three paired bronchoalveolar lavage (BAL) specimens with a 33% concordance. Five patients presented p53 exon VI mutations in biopsies and in two paired BALs with a 40% concordance. No mutations were found in p53 exon VII either in biopsies or in paired BAL samples with 100% concordance. Exon VIII mutations were found in six primary tumors and in two BALs with a 33% concordance. Of 36 patients, we detected 7 (19.4%) with K-ras exon I mutations on tumor samples. DGGE analysis of DNA from BAL samples revealed three mutations distributed on K-ras exon I with a 42% overall concordance with respect to tumor tissue. Molecular screening by DGGE of p53-amplified DNA from BAL had cumulative 46.6% sensitivity, 100% specificity, and 77.7% accuracy. DGGE K-ras detection showed 43% sensitivity, 100% specificity, and 88.8% test accuracy. The method proposed demonstrated to be specific, accurate, and at relatively low cost but limited by low sensitivity in detecting the presence of neoplastic cells in patients with resectable non-small cell lung cancer.     

Perception of 3D shape in context: Contrast and assimilation

Whereas integration of shape and surround is held to occur through cue-dependent representations, we show that both cue-invariant and cue-dependent representations are involved. A central hinged plane and larger flanking plane were defined by either binocular disparity or motion. In a ‘within-cue’ condition, shape and surround were defined by the same cue and in a ‘cross-cue’ condition they were defined by a different cue. Observers compared the dihedral angle of the central shape with a constant reference. When the central shape was defined by disparity, the surround stimuli invoked a contrast bias in the within-cue condition, but shape assimilation occurred in the cross-cue condition. When the central shape was defined by motion there were overall no significant results, but if a contrast bias was observed, it was in the within-cue condition where integration could occur through cue-dependent representations.     

Primary cilia sensitize endothelial cells for fluid shear stress.

Primary cilia are mechanosensors for fluid shear stress, and are involved in a number of syndromes and congenital anomalies. We identified endothelial cilia in areas of low shear stress in the embryonic heart. The objective of the present study was to demonstrate the role of primary cilia in mechanosensing. Ciliated embryonic endothelial cells were cultured from the heart, and non-ciliated cells from the arteries. Non-ciliated cells that were subjected to fluid shear stress showed significantly less induction of the shear marker Krüppel-Like Factor-2, as compared to ciliated cells. In addition, ciliated cells from which the cilia were chemically removed show a similar decrease in flow response. This shows that primary cilia sensitize endothelial cells for fluid shear stress. In addition, we targeted and stabilized the connection of the cilium to the cytoplasm by treatment with Colchicine and Taxol/Paclitaxel, respectively, and show that microtubular integrity is essential to sense shear stress.     

Preoperative cranial ultrasound findings in infants with major congenital heart disease

BACKGROUND: Advances in diagnostic testing and surgical techniques have resulted in reduced mortality in neonates with congenital heart disease (CHD) and a major concern for neurological morbidity in the presence of preoperative neurological injury. OBJECTIVES: To determine the incidence and nature of preoperative cerebral ultrasound abnormalities in neonates with major CHD and to examine the relationship between cerebral abnormalities and the type of CHD. METHODS: Retrospective study; inclusion criteria: (1) neonates with major CHD admitted to the NICU over a 3-y period, (2) gestational age >35 wk, (3) documented preoperative cranial ultrasound available; exclusion criteria: (1) small for gestational age, (2) other congenital anomalies and/or chromosomal abnormalities, (3) a 5-min Apgar score <7, (4) congenital infection. Cranial ultrasounds (CUS) were reviewed without knowledge of the cardiac defect. CHDs were categorized. RESULTS: Fifty of 108 neonates with CHD met the inclusion criteria. Twenty-one patients (42%) had abnormalities on CUS. Thirteen of these (26%) had widened ventricular and/or subarachnoid spaces, three (6%) lenticulostriate vasculopathy, one (2%) calcification in the basal nuclei, and four (8%) had acute ischaemic changes. Cerebral abnormalities occurred more frequently in patients with coarctation or hypoplastic left heart syndrome (HLHS) than transposition of the great arteries (TGA) (63% vs 14%; n.s.).CONCLUSION: There is a high incidence of preoperative cerebral ultrasound abnormalities in this group of neonates with major CHD.     

Paired, quantitative measurements of hepatitis B virus DNA in saliva, urine and serum of chronic hepatitis B patients

OBJECTIVES: Despite an abundance of epidemiological evidence for horizontal transmission of hepatitis B virus (HBV), the transmission route remains to be fully elucidated. In a new approach, we evaluated quantitative HBV DNA content in serum, saliva and urine as a first step in exploring possible modes of horizontal transmission. METHODS: In an outpatient setting of an academic hospital, paired serum, saliva and urine samples were collected from 150 chronically infected HBV patients. A validated HBV DNA TaqMan assay was used to quantitatively measure HBV DNA. RESULTS: Mean log HBV DNA in serum was 5.8 (range, undetectable to 10.0 log HBV DNA) copies/ml, 50% of the patients had an HBV DNA above 10 copies/ml in serum. Mean log HBV DNA level in saliva was 3.2 (range, undetectable to 7.5) copies/ml, 15% had an HBV DNA above 10 copies/ml in saliva. Mean log HBV DNA level in urine was 2.6 (range, undetectable to 5.4) copies/ml and 1% had an HBV DNA above 10 copies/ml in urine. A high, non-linear correlation was shown between HBV DNA in serum and saliva (Spearman's rho 0.82) and between serum and urine (Spearman's rho 0.74). CONCLUSIONS: The significant amounts of HBV DNA found in saliva and urine in chronic HBV patients with high viraemia in serum may have implications for the understanding of hepatitis B epidemiology. The potential infectivity of these body fluids may provide an explanation for the 20% of cases of infection obtained through horizontal transmission for which the origin of infection is yet unknown.     

Intractable congenital chylous ascites

A male newborn presented with congenital chylous ascites, the treatment of which is difficult and complicated due to persistent loss of chyle. The primary therapeutic goal is to reduce the lymph flow and to allow the leakage to heal naturally. Usually, conservative management--enteric rest with total parental feeding or an enteral diet with medium-chain triglycerides--can achieve this. A new treatment option is the use of octeotride, a somatostatin analogue. Both therapies failed in our patient, and surgical ligation of the leaking lymphatic duct was successful. Conclusion: The success of the surgical treatment depends on localizing the leaking lymphatic duct. Lymphatic imaging with lymphoscintigraphy and pre-operative administration of lipophilic dye can assist the surgeon.     

Results of arthrodesis in neuropathic feet

We describe the results of arthrodesis for the treatment of recurrent acute neuropathic bone disease in 24 feet and of chronic disease with deformity in 91 feet, undertaken between January 1984 and December 2003. All were due to leprosy. Correction of the deformity was achieved in 80 of 106 feet (76%) and fusion in 97 of 110 feet (88%). In the 24 feet in which recurrent neuropathic bone disease was the reason for surgery, 17 (71%) obtained stability while in seven (29%) symptoms recurred postoperatively. Complications were experienced following 58 of the 110 operations (53%). In patients presenting primarily with deformity with a minimum follow-up of two years (79 feet), there was a reduced frequency of ulceration in 40 (51%). Normal footwear could be worn by 32 patients (40%) after surgery, while 40 (51%) required a moulded insole. Arthrodesis of the ankle in the neuropathic foot due to leprosy has a good overall rate of success although the rate of complications is high.