A biochromatographic framework to evaluate the calcium effect on the antihypertensive molecule-human serum albumin binding

The Ca(2+) cation effect on the antihypertensive molecule binding on human serum albumin (HSA) was studied by biochromatography. The thermodynamic parameters corresponding to this binding were determined for a wide range of Ca(2+) concentration (x). For the two antihypertensive molecules under study, their binding to HSA can be divided into two Ca(2+) cation concentration regions due to a HSA phase transition. This result was confirmed by an enthalpy-entropy investigation. For a low x value (below x(c)=1.6 mmol l(-1)), the HSA cavity was in an ordered solid-like state leading to an increase in the interactions between the antihypertensive drugs and the HSA cavity and consequently, a solute-HSA affinity increase. For x above x(c), the HSA cavity was in a disordered solid-like state, implying a decrease in the antihypertensive drug-HSA binding.     

Diffusion imaging in obstructive hydrocephalus

BACKGROUND AND PURPOSE: Hydrocephalus causes transependymal resorption of spinal fluid that in turn produces periventricular interstitial edema. This study was performed to determine if diffusion imaging can demonstrate this interstitial edema in the periventricular region in patients with obstructive hydrocephalus and if it can be used to assess the treatment response. METHODS: Twenty-one patients with obstructive hydrocephalus were evaluated with MR diffusion imaging before and after treatment. The change in ventricular size was measured by using the frontal and occipital horn ratio. The signal intensity abnormalities in periventricular white matter were scored. Average diffusion constants (D(av)) in the periventricular white matter were measured before and after treatment and compared with normal values. Post-treatment resolution of MR imaging abnormalities and changes in ventricular volume were compared with changes in D(av). RESULTS: D(av) measured from periventricular white matter was increased in hydrocephalic patients compared with age-matched control subjects by a mean of 6.9% (P <.02). After treatment, D(av) decreased by an average of 6.0%: D(av) decreased in 11 patients (53%), it remained essentially unchanged in seven (33%), and it increased in three (14%). CONCLUSION: For patients with obstructive hydrocephalus, diffusion is usually increased in the periventricular white matter. Therefore, increased D(av) may be a clinically useful sign of hydrocephalus, and it may prove useful in cases with equivocal clinical or imaging findings. Measurement of D(av) may be valuable in assessing the treatment response in these patients because D(av) usually decreases toward normal levels with successful treatment.     

T1a breast carcinoma and the role of axillary dissection

HYPOTHESIS: Axillary dissection (AD) does not affect recurrence or survival in T1a breast cancer. DESIGN: Cohort study comparing patients who underwent AD and those who did not. SETTING: Provincial cancer agency. PATIENTS: Six hundred ninety-one women with pathologically diagnosed T1a tumors. MAIN OUTCOME MEASURES: Rates of axillary metastases stratified according to grade and lymphovascular and/or neural invasion, rates of relapse, and disease-specific survival. RESULTS: Grade 1, 2, and 3 tumors without lymphovascular and/or neural invasion had axillary nodal involvement rates of 0.7%, 7%, and 7.8% of patients, respectively; with lymphovascular and/or neural invasion, axillary nodes were involved in 9.1%, 39.3%, and 44.4%, respectively. No statistically significant differences were found between the cohorts in relapse rates (P =.70) or survival (P =.84). CONCLUSION: Higher tumor grade and lymphovascular and/or neural invasion increased the rate of nodal metastases in T1a tumors, but AD did not improve relapse rates or breast cancer-specific survival.     

Rapid prenatal diagnosis of Down syndrome using quantitative fluorescence in situ hybridization on interphase nuclei

OBJECTIVES: Presently, conventional cytogenetic analysis of metaphase chromosomes remains the reference approach in prenatal diagnosis. However, this method is labor-intensive and time-consuming. The first step toward the rapid identification of aneuploidies is achieved by interphase fluorescence in situ hybridization (FISH) with centromeric or locus-specific probes. Spot counting using this type of probes is a reliable approach, but is very time-consuming with some technical and biological limitations. In this study, we present a new FISH method using image cytometry for the detection of trisomy 21 within interphase nuclei. METHODS: The method is based on a comparative quantitation of the fluorescence signals emitted by whole chromosome 21 and 22 painting probes cohybridized on interphase nuclei. The chromosomal imbalance was determined with an automated image cytometer by detecting an abnormal ratio of both fluorescence emissions when compared with the ratio obtained in normal cells. RESULTS: Ten blood samples and twenty amniotic fluids were analyzed. Results from FISH and standard cytogenetics were compared and 100% correlation was achieved. CONCLUSIONS: This method, which enables an easy detection of chromosomal imbalances without a need for metaphase preparations, can be applied to the diagnosis of trisomy 21 and extended to other disorders with chromosomal imbalances. Compared to other interphase FISH techniques, it avoids spot-scoring difficulties.     

Lung cancer screening

Low-dose CT screening for lung cancer is a complex and controversial topic. This article reviews the history of lung cancer screening trials and addresses the principles and confounding biases associated with screening. Chest radiography was initially used for lung cancer screening in the 1970s. In the mid-1990s helical single-detector CT came into use, followed by helical multidetector CT, the current method of screening. Results from prevalence studies and a few single-arm incidence studies have raised concerns about overdiagnosis and the high rate of nodule detection. Follow-up studies and further investigation are needed. To this end, a randomized, controlled trial sponsored by the National Cancer Institute is underway to evaluate diseasespecific mortality.     

Provocative discography in patients after limited lumbar discectomy: A controlled, randomized study of pain response in symptomatic and asymptomatic subjects

STUDY DESIGN: This was a prospective observational study of patients with low back pain and those without after laminotomy and discectomy. OBJECTIVES: To determine, using a strict experimental design, the relative pain intensity response to provocative discography in symptomatic and asymptomatic subjects after lumbar discectomy for intervertebral disc herniation. BACKGROUND: Provocative discography frequently is used to evaluate persistent or recurrent low back pain syndromes in patients who have undergone posterior discectomy. The validity of interpreting painful injections during this procedure has not been critically assessed. The prevalence of significantly painful disc injections in a group with good outcomes after surgery is not known. Knowing the rates of significantly painful injections in asymptomatic patients after lumbar discectomy may clarify the meaning of painful injections in symptomatic patients. METHODS: From a cohort of 240 patients who had undergone single-level limited discectomy for sciatica, 20 asymptomatic volunteers were recruited for experimental three-level lumbar discography. Inclusion criteria required nearly perfect scores on standardized back pain rating instruments, no other spinal pathology, and normal psychometric screening. A control group of 27 symptomatic patients, after single-level discectomy with intractable low back pain syndrome, and without other spinal pathology, underwent discography. Seven patients in the control group had normal psychometric tests. Experienced raters who were blinded to control versus experimental status of the subjects scored the magnetic resonance imaging, discogram, psychometric tests, and discography videotapes of the subjects' pain behavior. RESULTS: There were 8 of 20 (40%) positive injections of discs that had previous surgery in the asymptomatic group and 17 of 27 (63%) positive injections in the symptomatic group. Specifically with regard to the symptomatic group, there were 3 of 7 (43%) positive injections (all concordant) in patients with normal psychometric scores, as compared with 14 of 20 (70%) positive injections (12 concordant) in patients with abnormal psychometric scores. Injections of discs that had previous surgery resulted in a mean pain score of 2.1 of 5 in the asymptomatic group, 2.1 in the symptomatic group with normal psychometric scores, and 3.4 in the symptomatic group with abnormal psychometric scores. Of the discs not treated with surgery, 2 were positive in the asymptomatic group (10%), 3 in 2 symptomatic subjects with normal psychological testing (29), and 18 in 13 symptomatic subjects with abnormal psychometric testing (76%). CONCLUSIONS: A high percentage of asymptomatic patients with normal psychometric testing who previously have undergone lumbar discectomy will have significant pain on injection of their discs that had previous surgery (40%). This is not significantly different from the experience of symptomatic patients with normal psychometric testing undergoing discography on discs that had previous surgery. Patients with abnormal psychological profiles have significantly higher rates of positive disc injections than either asymptomatic volunteers or symptomatic subjects with normal psychological screening.     

Mechanism of chronic obstructive uropathy: increased expression of apoptosis-promoting molecules

BACKGROUND: We have demonstrated that renal tubular and interstitial cells undergo pronounced apoptosis during the course of chronic obstructive uropathy (COU). Apoptosis is a complex cellular process consisting of multiple steps, each of which is mediated by families of related molecules. These families may include receptor/ligand molecules such as Fas, Fas ligand, tumor necrosis factor receptor-1 (TNFR-1), and TNF-related apoptosis inducing ligand (TRAIL); signal transduction adapter molecules such as Fas-associated death domain (FADD), TNFR-1 associated death domain (TRADD), receptor-interacting protein (RIP), Fas-associated factor (FAF), and Fas-associated phosphatase (FAP); or effector molecules such as caspases. However, the mechanism of tubular cell apoptosis, as well as the pathogenetic relevance of these apoptosis-related molecules in COU, remains poorly understood. METHODS: Kidneys were harvested from sham-operated control mice and mice with COU created by left ureter ligation sacrificed in groups of three at days 4, 15, 30, and 45. To detect apoptotic tubular and interstitial cells, in situ end labeling of fragmented DNA was performed. To detect the expression of apoptosis-related molecules, ribonuclease protection assay was used with specific antisense RNA probes for Fas, Fas ligand, TNFR-1, TRAIL, FADD, TRADD, RIP, FAF, FAP, and caspase-8. Immunostaining for Fas, Fas ligand, TRAIL, TRADD, RIP, and caspase-8 was also performed. To assess the role of these molecules in COU-associated renal cell apoptosis, the frequencies of apoptotic tubular and interstitial cells were separately quantitated for each experimental time point, and their patterns of variation were correlated with those of apoptosis-related molecules. RESULTS: The obstructed kidneys displayed increased apoptosis of both tubular and interstitial cells. Tubular cell apoptosis appeared at day 4 after ureter ligation, peaked (fivefold of control) at day 15, and decreased gradually until the end of the experiment. In contrast, interstitial cell apoptosis sustained a progressive increase throughout the experiment. Apoptosis was minimal at all experimental time points for control and contralateral kidneys. Compared with control and contralateral kidneys, the ligated kidneys displayed a dynamic expression of mRNAs for many apoptosis-related molecules, which included an up to threefold increase for Fas, Fas ligand, TNF-R1, TRAIL, TRADD, RIP, and caspase-8, and an up to twofold increase for FADD and FAP, but there was little change for FAF. These mRNAs increased between days 4 and 15, decreased until day 30, but then increased again until day 45. The rise and fall of mRNAs between days 4 and 30 paralleled a similar fluctuation in tubular cell apoptosis in that period. The subsequent increase of mRNAs was correlated with a continuous rise of interstitial cell apoptosis. We demonstrated a positive immunostaining for Fas and Fas ligand in the tubular cells at early time points as well as in interstitial inflammatory cells at later time points. Although increased expression of TRAIL, TRADD, RIP, and caspase-8 was noted in tubular cells, there was no staining for these molecules in interstitial cells. CONCLUSION: The current study documents a dynamic expression of several molecules that are known to mediate the most crucial steps of apoptosis. It implicates these molecules in COU-associated renal cell apoptosis and in the pathogenesis of this condition. It also lays the foundation for interventional studies, including genetic engineering, to evaluate the molecular control of apoptosis associated with COU.