Utility of Papanicolaou test in diagnosis of cervical lesions: a study in a tertiary care centre of western Uttar Pradesh

Background: Cervical cancer is the most common cancer among females in India. It is also one of the major causes of deaths among them. Papanicolaou (Pap) cytological test helps in detecting the early epithelial abnormalities in cervical cells. The morbidity and mortality due to cervical cancer has come down in countries with well-established screening programmes at national level. This study was conducted to emphasize the value of cervical screening as a tool for early detection of non-neoplastic and neoplastic lesions of cervix. Slide positivity rate was computed and clinico-pathological correlation was done.Methods: This cross-sectional study was conducted from January 2012 to December 2014 (3 years), on 2202 women aged between 20-70 years coming for a Pap smear examination in a tertiary care hospital in Bareilly, India. After staining with conventional Papanicolaou technique, all cases were classified as per Bethesda nomenclature (2001).Results: Out of 2202 cases, 85.16% slides were found to be negative for intraepithelial abnormalities and 14.84% slides were labelled as positive for epithelial lesions. 97.67% of high grade lesions were found in women more than 40 years. Smears with epithelial cell abnormality were found more common in older age groups whereas smears negative for epithelial cell abnormality were found more common in young age groups.Conclusion: Pap test is a cost effective cancer screening and is a simple method to detect various lesions of cervix, non-neoplastic as well as neoplastic. High risk screening programmes should be directed to all women >40 years. Pap test is especially useful to diagnose precancerous lesions of cervix, thereby early detection of these lesions & subsequent proper treatment can be helpful in prevention of cervical cancer.

Development of 2,4-D-resistant transgenics in Indian oilseed mustard (Brassica juncea)

Transgenic lines resistant to the herbicide 2,4- dichlorophenoxyacetic acid (2,4-D) were developed in mustard (Brassica juncea), a major oilseed crop grown in more than six million hectares of land in North India. The developed construct contained the tfdA gene, encoding the enzyme 2,4-D monooxygenase, cloned downstream to the 35S promoter along with a leader sequence from RNA4 of alfalfa mosaic virus (AMV leader sequence), for improved expression of the transgene in plant cells. Southern analysis of T0 transgenics confirmed six out of 24 transgenics to be single copy events, from both the flanks of T-DNA. Selfed progeny derived from single copy tfdA lines germinated normally and rooted in medium containing 2,4-D at concentrations as high as 2.5 mg l–1 compared to the wild-type seedlings which did not root even at a concentration of 0.5 mg l–1. The tfdA transgenic lines were also sprayed with commercially available 2,4-D herbicide at concentrations ranging from 10 to 1000 mg l–1 under field conditions. Wild type plants were affected by levels as low as 10 mg l–1 and were completely killed at a concentration of 50 mg l–1. The four transgenic lines tested in the study were resistant to herbicide concentration of 500 mg l–1. The available transgenic lines can be used for testing the potential of 2,4-D in weed control including the control of parasitic weeds (Orobanche spp) of mustard and for low-till cultivation of mustard

Estimation of genetic diversity among sugarcane (Saccharum species complex) clones

he experimental materials consisted of 36 sugarcane clones including two checks (Co Pant 97222 and Co Pant 3220). Analysis of variance revealed significant differences among all the clones for all the traits under study namely no. of millable canes, cane height, single cane weight, juice sucrose percent , purity percent , cane yield and CCS yield except cane thickness, juice brix and juice extraction percent. The divergence studies through Mahalanobis D2 statistics grouped the 36 genotypes into eleven clusters. The maximum numbers of genotypes (21) were grouped in clusterI and the lowest (1) in cluster VI,VII,VIII,IX,X and XI. Members of cluster VII and XI (46.48) were found to be genetically most diverse on the basis of their inter cluster difference as opposite to clusters I and II (10.77) which are closely related. Cane height contributed maximum (15.397%) towards genetic divergence followed by Single cane weight (14.762%) and no. of millable cane (13.016%). These characters were considered to be most important for the genetic diversity. Lowest contribution was made by juice purity percent (4.286%) followed by Cane thickness(7.301%),Juice extraction percent (7.619%). Genetic diversity is important for sustainable production since greater losses of characteristics in any population limits its chances of survival. Little to no genetic diversity makes crops extremely susceptible to widespread biotic and abiotic stresses. Genetic diversity can be assessed by Mahalanobis D2 statistic, which is a morphometric method and a powerful tool in quantifying the degree of divergence at genotypic level

Retransformation of a male sterile barnase line with the barstar gene as an efficient alternative method to identify male sterile-restorer combinations for heterosis breeding

We report in this study, an improved method for identifying male sterile-restorer combinations using the barnase-barstar system of pollination control for heterosis breeding in crop plants, as an alternative to the conventional line × tester cross method. In this strategy, a transgenic male sterile barnase line was retransformed with appropriate barstar constructs. Double transformants carrying both the barnase and barstar genes were identified and screened for their male fertility status. Using this strategy, 66-90% of fertile retransformants (restored events) were obtained in Brassica juncea using two different barstar constructs. Restored events were analysed for their pollen viability and copy number of the barstar gene. Around 90% of the restored events showed high pollen viability and ~30% contained single copy integrations of the barstar gene. These observations were significantly different from those made in our earlier studies using line (barnase) × tester (barstar) crosses, wherein only two viable male sterile–restorer combinations were identified by screening 88 different cross-combinations. The retransformation strategy not only generated several independent restorers for a given male sterile line from a single transformation experiment but also identified potential restorers in the T0 generation itself leading to significant savings in time, cost and labour. Single copy restored plants with high pollen viability were selfed to segregate male sterile (barnase) and restorer (barstar) lines in the T1 progeny which could subsequently be diversified into appropriate combiners for heterosis breeding. This strategy will be particularly useful for crop plants where poor transformation frequencies and/or lengthy transformation protocols are a major limitation

Comparative mapping of Brassica juncea and Arabidopsis thaliana using Intron Polymorphism (IP) markers: homoeologous relationships, diversification and evolution of the A, B and C Brassica genomes

Background: Extensive mapping efforts are currently underway for the establishment of comparative genomics between the model plant, Arabidopsis thaliana and various Brassica species. Most of these studies have deployed RFLP markers, the use of which is a laborious and time-consuming process. We therefore tested the efficacy of PCR-based Intron Polymorphism (IP) markers to analyze genome-wide synteny between the oilseed crop, Brassica juncea (AABB genome) and A. thaliana and analyzed the arrangement of 24 (previously described) genomic block segments in the A, B and C Brassica genomes to study the evolutionary events contributing to karyotype variations in the three diploid Brassica genomes. Results: IP markers were highly efficient and generated easily discernable polymorphisms on agarose gels. Comparative analysis of the segmental organization of the A and B genomes of B. juncea (present study) with the A and B genomes of B. napus and B. nigra respectively (described earlier), revealed a high degree of colinearity suggesting minimal macro-level changes after polyploidization. The ancestral block arrangements that remained unaltered during evolution and the karyotype rearrangements that originated in the Oleracea lineage after its divergence from Rapa lineage were identified. Genomic rearrangements leading to the gain or loss of one chromosome each between the A-B and A-C lineages were deciphered. Complete homoeology in terms of block organization was found between three linkage groups (LG) each for the A-B and A-C genomes. Based on the homoeology shared between the A, B and C genomes, a new nomenclature for the B genome LGs was assigned to establish uniformity in the international Brassica LG nomenclature code. Conclusion: IP markers were highly effective in generating comparative relationships between Arabidopsis and various Brassica species. Comparative genomics between the three Brassica lineages established the major rearrangements, translocations and fusions pivotal to karyotype diversification between the A, B and C genomes of Brassica species. The inter-relationships established between the Brassica lineages vis-à-vis Arabidopsis would facilitate the identification and isolation of candidate genes contributing to traits of agronomic value in crop Brassicas and the development of unified tools for Brassica genomics

Identification and quantification of biological active constituents of Amritarishta, a herbal formulation

Herbal formulations have been used by Indian and Chinese traditional systems of medication for a long time. Amritarishtais one of the herbal formulations that possess various biological activity viz., antioxidant, anticancer, analgesic, antipyretic,antidiabetic, etc. The active constituents include gallic acid, tannic acid, piperine, and quercetin, etc. Ethanolic extract of theformulation was analysed and quantified. Rf (Retardation factor), functional groups and amount of some of the major chemicalconstituents were analysed by TLC, FTIR, LC/MS, HPTLC and HPLC, respectively. LC/MS results reveal the presence ofquercetin, piperine, tannic acid and gallic acid in the formulation. With the help of HPTLC and HPLC, the quantity of 4chemical constituents in the formulation was estimated. This type of study is completely new to herbal research

Cyberknife fractionated radiotherapy for adrenal metastases: Preliminary report from a multispecialty Indian cancer care center

Purpose: Metastasis to adrenal gland from lung, breast, and kidney malignancies are quite common. Historically radiotherapy was intended for pain palliation. Recent studies with stereotactic body radiotherapy (SBRT) including Cyberknife robotic radiosurgery aiming at disease control brings about encouraging results. Here we represent the early clinical experience with Cyberknife stereotactic system from an Indian cancer care center. The main purpose of this retrospective review is to serve as a stepping stone for future prospective studies with non- invasive yet effective technique compared to surgery. Methods: We retrospectively reviewed four cases of adrenal metastases (three: lung and one: renal cell carcinoma) treated with Cyberknife SBRT. X sight spine tracking was employed for planning and treatment delivery. Patients were evaluated for local response clinically as well as with PETCT based response criteria.Results: With a median gross tumor volume of 20.5 cc and median dose per fraction of 10 Gy, two patients had complete response (CR) and two had partial response (PR) when assessed 8-12 weeks post treatment as per RECIST. There was no RTOG grade 2 or more acute adverse events and organs at risk dosage were acceptable. Till last follow up all the patients were locally controlled and alive. Conclusion: Cyberknife SBRT with its unique advantages like non- invasive, short duration outpatient treatment technique culminating in similar local control rates in comparison to surgery is an attractive option. World literature of linear accelerator based SBRT and our data with Cyberknife SBRT with small sample size and early follow up are similar in terms of local control in adrenal metastases. Future prospective data would reveal more information on the management of adrenal metastases

Preliminary experience of fractionated stereotactic radiosurgery with extend system of Gamma Knife

Purpose: The purpose of this study is to present multisession stereotactic radiosurgery with initial experience using custom made extend system (ES) of Gamma Knife.Methods: The ES is comprised of a carbon fiber frame also called extend frame, vacuum head rest cushion, patient surveillance unit and a configurable front piece with dental impression tray. The extend frame is a rigid connection between patient's head and patient positioning system (PPS) of Gamma Knife. A dental impression of patient was created and attached to the frontal piece of extend system. The treatment setup involves positioning the patient within the extend frame using patient specific headrest cushion and front piece. The reference patient’s head position was recorded through measurements of repositioning check tool (RCT) apertures using a high precision digital probe before computed tomography (CT) scan. The RCT measurements taken before treatment were compared with recorded reference position to ensure appropriate patient treatment position. Volumetric magnetic resonance (MR) scan was co-registered with stereotactic CT scan on Leksell Gamma plan. Fused MR to CT images on Gamma Plan was utilized to delineate regions of interest and prepare a precise treatment plan. The presented study includes positional reproducibility check and dosimetric evaluation of ten patients treated with ES.Results: Forty-three fractions on ten patients with prescribed treatment format were delivered successfully. An average tumor volume of 11.26 cm3 (range, 340 mm3 to 59.12 cm3) was treated with ES. The mean tumor coverage of 91.91% (range, 90% to 95%) was able to achieve at 50% prescription isodose without compromising adjacent normal structure radiation dose tolerances. The mean inter-fraction positional variation of 0.69 mm influences an inherent strength of immobilization technique. Follow-up of seven patients at a median interval of 16 months (range, 9 months to 26 months) showed evidence of 100% radiographic control with improved clinical results.Conclusion: Conjugative clinical outcome shows the efficacy of fractionation in various clinical indications