Multicolour fluorescence in situ hybridization analysis of t(14;18)-positive follicular lymphoma and correlation with gene expression data and clinical outcome

In order fully to identify secondary chromosomal alterations, such as duplications, additions and marker chromosomes that remained unresolved by G banding, 60 cases of t(14;18)-positive follicular lymphoma (FL) were analysed by multicolour karyotyping techniques [multicolour fluorescence in situ hybridization (MFISH)/multicolour banding for chromosome 1 (MBAND1)]. A total of 165 additional structural chromosomal aberrations were delineated. An increased frequency of chromosomal gains involving X, 1q, 2, 3q27-q29, 5, 6p11-p21, 7, 8, 11, 12, 14q32, 17q, 18 and 21 and deletions of 1p36, 3q28-q29, 6q, 10q22-q24 and 17p11-p13 was revealed by the MFISH/MBAND1 analysis. Balanced translocations other than t(14;18) were uncommon, whereas unbalanced translocations were numerous. Deletion of 1p36 and duplication of 1p33-p35, 1p12-p21 and 1q21-q41 were regularly involved in chromosome 1 alterations, seen in 53% of the cases. A strong correlation was demonstrated between gains of individual chromosomal bands and increased gene expression, including 1q22/MNDA, 6p21/CDKN1A, 12q13-q14/SAS, 17q23/ZNF161, 18q21/BCL2 and Xq13/IL2RG. Unfavourable overall survival was associated with del(1)(p36) and dup(18q). These data support the notion that translocation events are primarily responsible for FL disease initiation, whereas the unbalanced chromosomal gains and losses that mirror the gene expression patterns characterize clonal evolution and disease progression, and thus provide further insights into the biology of FL.     

Hepatitis C virus replication is directly inhibited by IFN-alpha in a full-length binary expression system

Hepatitis C virus (HCV) is a leading cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. The absence of culture systems permissive for HCV replication has presented a major bottleneck to antiviral development. We sought to recapitulate the early steps in the life cycle of HCV by means of DNA-based expression of viral genomic sequences. Here we report expression of replicating HCV RNA by using a, to our knowledge, novel binary expression system in which cells were transfected with a T7 polymerase-driven full-length HCV cDNA plasmid containing a cis-acting hepatitis Delta ribozyme to control 3' cleavage, and infected with vaccinia-T7 polymerase. HCV genomic and replicative strand synthesis, in addition to protein synthesis, was detectable and depended on full-length HCV sequences. Moreover, the system was capable of generating HCV RNA quasispecies, consistent with the action of the low-fidelity HCV NS5B RNA polymerase. IFN-alpha, but not ribavirin, directly inhibited the viral replicative cycle in these cells, identifying the virus itself and not solely the immune system as a direct target of IFN action. The availability of a cell-based test for viral replication will facilitate screening of inhibitory compounds, analysis of IFN-resistance mechanisms, and analysis of virus-host cell interactions.     

Sonographic Prediction of Scar Dehiscence in Women with Previous Cesarean Section

Purpose

To estimate the risk of uterine dehiscence/rupture in women with previous cesarean section (CS) by comparing the thickness of lower uterine segment (LUS) and myometrium with trans-abdominal (TAS) and trans-vaginal sonography (TVS).

Method

In this case-control study, in 100 pregnant women posted for elective CS (with or without previous CS; group 1 and group 2 respectively), the thickness of LUS and myometrium was measured sonographically (TAS and TVS). Intra-operatively, LUS was graded (grades I–IV), and its thickness was measured with calipers. The primary outcome of the study was correlation between echographic measurements (TAS and TVS) and features of LUS (grades I–IV) at the time of CS. Secondary outcomes were correlation between myometrial thickness, number of previous CS, and inter-delivery interval with LUS (grades I–IV).

Results

Sonographic measurements of LUS and myometrium were significantly different between the two groups (both TAS and TVS p value = 0.000 each). However, the number of previous CS (p = 0.440) and inter-delivery interval (p = 0.062) had no statistically significant correlation with thickness of LUS.

Conclusions

Sonographic evaluation of LUS scar and myometrial thickness (both with TAS and TVS) is a safe, reliable, and non-invasive method for predicting the risk of scar dehiscence/rupture. Specific guidelines for TOLAC, after sonographic assessment of women with previous CS, are need of the hour.Keyword: Transabdominal ultrasonography, Transvaginal ultrasonography, Cesarean section, Pregnancy, Cesarean scar     

HELLP or Help: A Real Challenge

Objectives

To ascertain the prevalence, presentation, diagnosis, severity, and complications of HELLP syndrome.

Materials and Methods

This is a prospective observational study analyzing the conditions and the data of 24 cases of HELLP syndrome in a tertiary care hospital. The analysis was done for the demographic characteristics, presentation of these patients, complications associated, and the perinatal outcome.

Results

0.45 % of the patients admitted for delivery developed HELLP syndrome. Majority of the patients developed the condition in 30–36 weeks period of gestation, while five patients developed it in the postpartum period. The condition led to 12.5 % of maternal and 45.8 % of perinatal mortality.

Conclusion

HELLP syndrome is an important cause for maternal and perinatal morbidity and mortality.     

Benign recurrent intrahepatic cholestasis (BRIC) in an adult

Benign recurrent intrahepatic cholestasis is a rare hereditary disorder characterised by recurrent episodes ofcholestasis. We report the case of a young male patient with benign recurrent intrahepatic cholestasis who presented to us with recurrent cholestatic jaundice and pruritus with negative work up for all possible aetiologies and a liver biopsy consistent with intrahepatic cholestasis. He improved on treatment with ursodeoxycholic acid and ondansterone and is doing well on follow up.     

Angioplasty Guidewire Velocity: A New Simple Method to Calculate Absolute Coronary Blood Velocity and Flow

The Thrombolysis In Myocardial Infarction (TIMI) frame count is a relative index of coronary flow that measures time by counting the number of frames required for dye to travel from the ostium to a standardized coronary landmark in a cineangiogram filmed at a known speed (frames/s). We describe a new method to measure distance along arteries so that absolute velocity (length ÷ time) and absolute flow (area × velocity) may be calculated in patients undergoing percutaneous transluminal coronary angiography (PTCA). After PTCA, the guidewire tip is placed at the coronary landmark and a Kelly clamp is placed on the guidewire where it exits the Y-adapter. The guidewire tip is then withdrawn to the catheter tip and a second Kelly clamp is placed on the wire where it exits the Y-adapter. The distance between the 2 Kelly clamps outside the body is the distance between the catheter tip and the anatomic landmark inside the body. Velocity (cm/s) may be calculated as this distance (cm) ÷ TIMI frame count (frames) × film frame speed (frames/s). Flow (ml/s) may be calculated by multiplying this velocity (cm/s) and the mean cross-sectional lumen area (cm²) along the length of the artery to the TIMI landmark. In 30 patients, velocity increased from 13.9 ± 8.5 cm/s before to 22.8 ± 9.3 cm/s after PTCA (p <0.001). Despite TIMI grade 3 flow both before and after PTCA in 18 patients, velocity actually increased 38%, from 17.0 ± 5.4 to 23.5 ± 9.0 cm/s (p = 0.01). For all 30 patients, flow doubled from 0.6 ± 0.4 ml/s before to 1.2 ± 0.6 ml/s after PTCA (p <0.001). In the 18 patients with TIMI grade 3 flow both before and after PTCA, flow increased 86%, from 0.7 ± 0.3 to 1.3 ± 0.6 ml/s (p = 0.001). Distance along coronary arteries (length) can be simply measured using a PTCA guidewire. This length may be combined with the TIMI frame count to calculate measures of absolute velocity and flow that are sensitive to changes in perfusion. TIMI grade 3 flow is composed of a range of velocities and flows.