Comparative evaluation of perimenopausal abnormal uterine bleeding by transvaginal sonography, hysteroscopy and endometrial biopsy

To evaluate the causes of abnormal uterine bleeding in perimenopausal women and to achieve the greatest diagnostic accuracy with the least risk, a cross-sectional study was done among 85 patients between the ages 40 and 55 years. Transvaginal sonography, hysteroscopy and histopathological examination of endometrium were done in all the cases. Kappa statistics was calculated to show the agreement between the different investigations. Considering histopathological report to be the gold standard, for diagnosis of hyperplastic endometrium, transvaginal sonography and hysteroscopy showed fair agreement (k=0.34) and good agreement (k=0.51) and hysteroscopic diagnosis of polyp showed strong agreement (k=0.81). Sensitivity, specificity, positive predictive value, negative predictive value for diagnosis of hyperplastic endometrium were 43.75%, 95.65%, 70% and 88% respectively whereas in polyp by transvaginal sonography they were 50%, 89.16%, 100%, 98.67% respectively; by hysteroscopy these were 50%, 95.78%, 70%, 90.36% respectively in hyperplasia and 71.43%, 100%, 100%, 94.67% respectively in polyp. Transvaginal sonography is most important for diagnosis of anatomical lesion. Hysteroscopy is most specific and sensitive for diagnosis of polyp but less specific for endometrial hyperplasia.     

Characterization and Studies of the Cellular Interaction of Native Colonization Factor CS6 Purified from a Clinical Isolate of Enterotoxigenic Escherichia coli

CS6 is a widely expressed colonization factor of enterotoxigenic Escherichia coli (ETEC). To date, CS6 has not been well characterized in its native state. Here, we purified CS6 for the first time from an ETEC clinical isolate. Purified CS6 was composed of two structural subunits, CssA and CssB, which were present in equal amounts and tightly linked through noncovalent, detergent-stable association. The CssA subunit was poorly immunogenic, whereas CssB was highly immunogenic. Although the predicted molecular mass of CssA is 15 kDa, the purified CssA has an effective molecular mass of 18.5 kDa due to fatty acid modification. When purified CS6 was screened for its ability to bind with different extracellular matrix proteins, fibronectin (Fn) was found to interact with CS6 as well as CssA in a dose-dependent and saturable manner. This interaction was inhibited both by a synthetic peptide corresponding to the C-terminal hydrophilic, surface-exposed region of CssA (positions 112 to 126) and by the antibody derived against this region. Enzyme-linked immunosorbent assay results showed that CssA interacted with the 70-kDa N-terminal domain of Fn. The modifications on CssA probably do not play a role in Fn binding. Preincubation of INT 407 cells with CssA, but not CssB, inhibited ETEC binding to these cells. The results suggested that CS6-expressing ETEC binds to Fn of INT 407 cells through the C-terminal region of CssA. Purified CS6 was found to colocalize with Fn along the junctions of INT 407 cells. Based on the results obtained, we propose that CS6-expressing ETEC binds to the intestinal cells through Fn for colonization.Enterotoxigenic Escherichia coli (ETEC) infection is the leading cause of infantile diarrhea in developing countries and an important etiologic agent for traveler''s diarrhea. ETEC accounts for approximately 210 million diarrhea episodes and 380,000 deaths annually (35). Community-based studies conducted in developing countries with children younger than 5 years have shown that ETEC was the most frequently isolated enteropathogen (34, 35). As a cause of traveler''s diarrhea, ETEC was found to be associated with 40 to 70% of the cases, with drastic outcome in terms of morbidity and economic consequences (34).In order to initiate pathogenesis, ETEC strains must adhere to the small intestine (14). This event is mediated by several proteinaceous surface antigens, collectively known as colonization factor antigens (CFAs) (6). To date, more than 25 distinct colonization factors have been identified, of which CS6 is the most prevalent in many countries (7, 20, 22). Many of the colonization factors have morphology of fimbriae or pili (14). However, the morphology of CS6 has not so far been defined. CS6 was assumed to be either a nonfimbrial or a short oligomeric assembly that does not protrude enough to be visualized under an electron microscope (17). Functional CS6 is expressed and transported to the bacterial surface in a chaperone-usher pathway. CssC and CssD are the chaperone and usher proteins, respectively, that help surface expression of the CS6 structural subunits, CssA and CssB (33).The role of CS6 in intestinal adherence has been demonstrated using CS6-expressing whole bacteria, but the receptor specificity is still unknown (11). A recent report has shown that when CssB is mutated, binding of bacteria to a colonic cell line (CaCo-2) is reduced slightly compared to that of the bacteria expressing whole CS6 (30).Here, we have purified CS6 to homogeneity from a clinical isolate of ETEC and separated its subunits (CssA and CssB) for the first time. We have characterized CS6 in its native form and demonstrated that fibronectin (Fn) is the interacting matrix for adherence. The carboxy-terminal (C-terminal) region of CssA plays a key role in this interaction with the amino-terminal (N-terminal) region of Fn.     

Seizure, spinal schwannoma, peripheral neuropathy and pulmonary stenosis �C A rare combination in a patient of Neurofibromatosis 1

Neurofibromatosis 1 (NF1) is the most common neurocutaneous syndrome. It is estimated to occur in approximately 1 out of every 3300 infants. The manifestations of this condition are diverse and can arise from almost any system in the body. The neurofibroma is the hallmark lesion of NF1 that develops from peripheral nerves. Here, we are reporting an 18-year-old girl with NF1. Clinical diagnosis was made according to the diagnostic criteria established by the National Institutes of Health Consensus Development Conference in 1987. She presented with quadriparesis due to dumbbell-shaped spinal schwannoma in the cervical region. She had history of recurrent seizures in the past, with poor scholastic performance. There were clinical and electrophysiological features of peripheral neuropathy and clinical and echocardiographical features of pulmonary stenosis. These are uncommon features of NF 1. The presence of all these features in a single patient makes it a unique case.     

A modified spreader for pyloromyotomy

A modified spreader for pyloromyotomy is innovated for congenital hypertrophic pyloric stenosis. It was found safe, effective and useful.     

Involvement of ROS in chlorogenic acid-induced apoptosis of Bcr-Abl CML cells

Chlorogenic acid (Chl) has been reported to possess a wide range of biological and pharmacological properties including induction of apoptosis of Bcr-Abl⁺ chronic myeloid leukemia (CML) cell lines and clinical leukemia samples via inhibition of Bcr-Abl phosphorylation. Here we studied the mechanisms of action of Chl in greater detail. Chl treatment induced an early accumulation of intracellular reactive oxygen species (ROS) in Bcr-Abl⁺ cells leading to downregulation of Bcr-Abl phosphorylation and apoptosis. Chl treatment upregulated death receptor DR5 and induced loss of mitochondrial membrane potential accompanied by release of cytochrome c from the mitochondria to the cytosol. Pharmacological inhibition of caspase-8 partially inhibited apoptosis, whereas caspase-9 and pan-caspase inhibitor almost completely blocked the killing. Knocking down DR5 using siRNA completely attenuated Chl-induced caspase-8 cleavage but partially inhibited apoptosis. Antioxidant NAC attenuated Chl-induced oxidative stress-mediated inhibition of Bcr-Abl phosphorylation, DR5 upregulation, caspase activation and CML cell death. Our data suggested the involvement of parallel death pathways that converged in mitochondria. The role of ROS in Chl-induced death was confirmed with primary leukemia cells from CML patients in vitro as well as in vivo in nude mice bearing K562 xenografts. Collectively, our results establish the role of ROS for Chl-mediated preferential killing of Bcr-Abl⁺ cells.     

Misoprostol for termination of mid-trimester post-Caesarean pregnancy

OBJECTIVE: To evaluate the efficacy and safety of PGE1 analogue, misoprostol, for inducing abortion or labour during mid-trimester in women who have had a prior Caesarean section (one or more). STUDY DESIGN: Women who had to undergo termination of pregnancy between 13 and 26 weeks of gestation for various indications and who had at least one previous Caesarean section were studied over a period of two and a half years. The standard regimen for misoprostol in all the cases was 400 microg up to 20 weeks of gestation and 200 microg for pregnancies longer than 20 weeks, either vaginally or sublingually every six hours (up to maximum 24 h). A contemporaneous cohort of women undergoing the same procedure for similar indications but without scarred uteri served as control. RESULTS: Eighty women in the study group underwent termination procedures for unwanted pregnancy, missed abortion, fetal anomaly or fetal death. The median induction-abortion interval was 16.4 h (10-21 h) and did not differ much from that in women without previous Caesarean delivery (median: 15.6 h; range 9.6-20 h), P > 0.05. Misoprostol was found to be safe in our cohort of post-Caesarean women and there was no case of scar rupture or dehiscence. No significant differences in rates of incomplete abortions, blood loss or sepsis were detected in the study group compared to the control group. CONCLUSION: The use of misoprostol for mid-trimester pregnancy termination is not contraindicated in women with Caesarean scar and is effective and comparable with those in women without scarred uteri.     

Hydroxychavicol, a Piper betle leaf component, induces apoptosis of CML cells through mitochondrial reactive oxygen species-dependent JNK and endothelial nitric oxide synthase activation and overrides imatinib resistance

Alcoholic extract of Piper betle (Piper betle L.) leaves was recently found to induce apoptosis of CML cells expressing wild type and mutated Bcr-Abl with imatinib resistance phenotype. Hydroxy-chavicol (HCH), a constituent of the alcoholic extract of Piper betle leaves, was evaluated for anti-CML activity. Here, we report that HCH and its analogues induce killing of primary cells in CML patients and leukemic cell lines expressing wild type and mutated Bcr-Abl, including the T315I mutation, with minimal toxicity to normal human peripheral blood mononuclear cells. HCH causes early but transient increase of mitochondria-derived reactive oxygen species. Reactive oxygen species-dependent persistent activation of JNK leads to an increase in endothelial nitric oxide synthase-mediated nitric oxide generation. This causes loss of mitochondrial membrane potential, release of cytochrome c from mitochondria, cleavage of caspase 9, 3 and poly-adenosine diphosphate-ribose polymerase leading to apoptosis. One HCH analogue was also effective in vivo in SCID mice against grafts expressing the T315I mutation, although to a lesser extent than grafts expressing wild type Bcr-Abl, without showing significant bodyweight loss. Our data describe the role of JNK-dependent endothelial nitric oxide synthase-mediated nitric oxide for anti-CML activity of HCH and this molecule merits further testing in pre-clinical and clinical settings.     

PCR-based identification of common colonization factor antigens of enterotoxigenic Escherichia coli

Colonization factor antigens (CFAs) of enterotoxigenic Escherichia coli (ETEC) have been classified into several groups based on their distinct antigenicity. We describe here a PCR-based method to detect common CFAs of ETEC, which were characterized using conventional serology. This PCR assay is simple and sensitive for the detection of expressed CFA genes.