Mutations in coagulation factor XIII A gene in eight unrelated Indians. Five novel mutations identified by a novel PCR-CSGE approach

Factor XIII deficiency is a rare autosomal (1:2,000,000) recessive disorder of blood coagulation usually attributed to mutations in the coagulation factor XIII (FXIII) A gene. We have studied the molecular basis of FXIII deficiency in eight unrelated South Indian patients. Their diagnosis was based on clinical history, normal plasma clotting times and increased solubility of fibrin clot in 5 mol/l urea. Genomic DNA was screened for FXIII A gene defects by a novel PCR and CSGE strategy. Mutations were identified in all these patients. Five of these were novel mutations occurring in four patients. These included a novel c.210T > G transversion in homozygosity in exon 3 predicting a Tyr69X in the beta-sandwich domain in one patient. Another patient was compound heterozygote for a novel c.791C > T transition predicting a Ser263Phe in the core domain and a novel c.2045-1G > A transition at the acceptor splice junction of intron 14. Two novel frame shifts were also identified in two patients in a homozygous condition. One of them resulted from a single base 'G' duplication (c.892_895dupG) at codons Ser290/Ala291fs affecting the core domain and the other was due to a single base 'A' duplication (c.1642_1644dupA) and at codonTyr547fs affecting barrel-1 domain. The remaining four patients had the previously reported Arg260His, Ser413Leu, and Val414Phe (n = 2) missense mutations in the core domain. The novel mutations identified were considered to be disease causative by studying the nature of mutation, the degree of conservation of the mutated aminoacid among transglutaminases of different species and by molecular modeling. Apart from describing a significant number of novel mutations, this report is the first study from Southern India to describe FXIII A gene mutations.     

Acanthus ilicifolius plant extract prevents DNA alterations in a transplantable Ehrlich ascites carcinoma-bearing murine model

AIM: To investigate the chemopreventive efficacy of the Indian medicinal plant Acanthus ilicifolius L Acanthaceae in a transplantable Ehrlich ascites carcinoma (EAC)- bearing murine model. METHODS: Male Swiss albino mice were divided into four groups: Group A was the untreated normal control; Group B was the EAC control mice group that received serial, intraperitoneal (ip) inoculations of rapidly proliferating 2 x 105 viable EAC cells in 0.2 mL of sterile phosphate buffered saline; Group C was the plant extract-treated group that received the aqueous leaf extract (ALE) of the plant at a dose of 2.5 mg/kg body weight by single ip injections, once daily for 10, 20 and 30 consecutive days following tumour inoculation (ALE control); and Group D was the EAC ALE- treatment group. The chemopreventive potential of the ALE was evaluated in a murine model by studying various biological parameters and genotoxic markers, such as tumour cell count, mean survival of the animals, haematological indices, hepatocellular histology, immunohistochemical expression of liver metallothionein (MT) protein, sister-chromatid exchanges (SCEs), and DNA alterations.RESULTS: Treatment of the EAC-bearing mice with the ALE significantly (P < 0.001) reduced viable tumour cell count by 68.34% (228.7 x 106 ± 0.53) when compared to EAC control mice (72.4 x 106 ± 0.49), and restored body and organ weights almost to the normal values. ALE administration also increased (P < 0.001) mean survival of the hosts from 35 ± 3.46 d in EAC control mice to 83 ± 2.69 d in EAC ALE-treated mice. Haematological indices also showed marked improvement with administration of ALE in EAC-bearing animals. There was a significant increase in RBC count (P < 0.001), hemoglobin percent (P < 0.001), and haematocrit value (P < 0.001) from 4.3 ± 0.12, 6.4 ± 0.93, and 17.63 ± 0.72 respectively in EAC control mice to 7.1 ± 0.13, 12.1 ± 0.77, and 30.23 ± 0.57 respectively in EAC ALE-treated group, along with concurrent decrement (P < 0.001) in WBC count from 18.8 ± 0.54 in EAC control to 8.4 ± 0.71 in EAC ALE. Furthermore, treatment with ALE substantially improved hepatocellular architecture and no noticeable neoplastic lesions or foci of cellular alteration were observed. Daily administration of the ALE was found to limit liver MT expression, an important marker of cell proliferation with concomitant reduction in MT immunoreactivity (62.25 ± 2.58 vs 86.24 ± 5.69, P < 0.01). ALE was also potentially effective in reducing (P < 0.001) the frequency of SCEs from 14.94 ± 2.14 in EAC control to 5.12 ± 1.16 in EAC ALE-treated group. Finally, in comparison to the EAC control, ALE was able to suppress in vivo DNA damage by abating the generations of ‘tailed’ DNA by 53.59% (98.65 ± 2.31 vs 45.06 ± 1.14, P < 0.001), and DNA single-strand breaks (SSBs) by 38.53% (3.14 ± 0.31 vs 1.93 ± 0.23, P < 0.01) in EAC-bearing murine liver. CONCLUSION: Our data indicate that, ALE is beneficial in restoring haematological and hepatic histological profiles and in lengthening the survival of the animals against the proliferation of ascites tumour in vivo. Finally, the chemopreventive efficacy of the ALE is manifested in limiting MT expression and in preventing DNA alterations in murine liver. The promising results of this study suggest further investigation into the chemopreventive mechanisms of the medicinal plant A. ilicifolius in vivo and in vitro.     

Study of Prevalence, Awareness,and control of Hypertension in a Suburban Area of Kathmandu, Nepal

BACKGROUND, In the context of rapidly raising occurrence of cardiovascular diseases in the developing countries, it becomes imperative to study the scenario in its various aspects. The present study in Nepal deals with the hypertension as it is one of the major risk factors of cardiovascular diseases. METHODS AND RESULTS, A house-to-house survey was conducted in a suburban area of Kathmandu valley from February to June 2005 in adult population (age >/=18 years) to estimate the prevalence, awareness, treatment, and control rates of hypertension. Blood pressure was measured twice using standardized mercury sphygmomanometer, and an average of the two readings was taken. Total number of subjects were 1114 (men:541; women: 573; mean age: 37.8 -/+ 16.3 years). Overall prevalence of hypertension was 19.7% (22.2% in men and 17.3% in women, p < 0.05).Prevalence of hypertension in age group of >/=40 years was 36%.Awareness, treatment, and control rates were 41.1%, 26%, and 6%, respectively. CONCLUSION, Our study indicates that prevalence of hypertension is significant in Nepal and is comparable with other developing countries of this region. Awareness, treatment, and control rates are poor.     

Intraoperative arcuate transverse keratotomy with phacoemulsification

PURPOSE: To evaluate the efficacy of paired intraoperative arcuate transverse keratotomy at a 7-mm-diameter zone along with a 3.5-mm clear corneal phaco tunnel in the steeper axis to correct pre-existing astigmatism. METHODS: A prospective randomized case-control study was conducted on 34 eyes of 28 patients with immature senile cataract. They were divided into two groups; in one group (17 eyes) intraoperative arcuate keratotomy was coupled with phacoemulsification in the steeper meridian (arcuate keratotomy group; mean preoperative astigmatism 2.28 +/- 0.89 D) and the other group (17 eyes) phacoemulsification was performed in the steeper meridian without arcuate keratotomy (control group; mean preoperative astigmatism 2.04 +/- 0.50 D). The patients were examined at 1 day, and 1, 4, and 8 weeks postoperatively. Correction of keratometric astigmatism, surgically induced refractive changes, magnitude and axis of cylinder, spherical equivalent refraction, with and against the wound change, and coupling ratio were evaluated. RESULTS: Mean reduction in keratometric astigmatism in the keratotomy group was 1.26 +/- 0.54 D (P = .0067) and in the control group was 0.48 +/- 0.60 D (P = .0423). The difference in reduction of keratometric astigmatism between the two groups was statistically significant (P = .0296). Surgically induced refractive change at 8 weeks follow-up was 2.15 +/- 1.13 D in the keratotomy group and 1.50 +/- 1.32 D in the control group (P = .046). Coupling ratio was -1.10 +/- 0.43 in the keratotomy group at 8 weeks after surgery while the control group was -0.82 +/- 0.38. CONCLUSION: A combination of intraoperative arcuate keratotomy with steep axis phacoemulsification incision is more effective than steep axis phacoemulsification incision alone in reducing pre-existing astigmatism.     

An endoscopic,cadaveric analysis of the roof of the fourth ventricle

We performed endoscopic dissections of the roof of the fourth ventricle in eight fresh human cadaveric heads to characterize the endoscopic anatomy of the roof of the fourth ventricle and the anatomical configuration of the structures forming its roof. We also made three-dimensional (3D) silicone casts of the fourth ventricle in seven formalin-fixed specimens to evaluate the 3D configuration of the structures that create the roof of the fourth ventricle. The roof of the fourth ventricle can be divided into three zones. The upper zone is formed by the superior cerebellar peduncle and superior medullary velum and is associated with the lingula. The middle zone is formed by the inferior cerebellar peduncles and inferior medullary velum and is associated with the nodule in the midline and with the peduncle of the flocculus. The lower zone is formed by the tela choroidea and is associated with the tonsils. The 3D shape of the roof the fourth ventricle resembles that of a rhomboid-based pyramid; the edges of the base represent the borders of the ventricle, and the apex is the cerebellar fastigium. The lateral recess is shaped like a triangular-based pyramid, with its base connected to the cavity of the fourth ventricle and its tip opening into the lateral cerebellomedullary cistern through the foramen of Luschka. Our results may help in the endoscopic exploration of and microsurgical approaches to the fourth ventricle through its roof.