Indigenously developed multipurpose acrylic head phantom for verification of IMRT using film and gel dosimetry.

The purpose of this study was to validate the newly designed acrylic phantom for routine dosimetric purpose in radiotherapy. The phantom can be used to evaluate and compare the calculated dose and measured dose using film and gel dosimetric methods. In this study, a doughnut-shaped planning target volume (8.54 cm3) and inner organ at risk (0.353 cm3) were delineated for an IMRT test plan using the X-ray CT image of the phantom. The phantom consists of acrylic slabs which are integrated to form a human head with a hole in the middle where several dosimetric inserts can be positioned for measurement. An inverse planning with nine coplanar intensity-modulated fields was created using Pinnacle TPS. For the film analysis, EBT2 film, flatbed scanner, in-house developed MATLAB codes and ImageJ software were used. The 3D dose distribution recorded in the MAGAT gel dosimeter was read using a 1.5 T MRI scanner. Scanning parameters were CPMG pulse sequence with 8 equidistant echoes, TR = 5600, echo step = 22 ms, pixel size = 0.5 × 0.5, slice thickness = 2 mm. Using a calibration relationship between absorbed dose and spin-spin relaxation rate (R2), R2 images were converted to dose images. The dose comparison was accomplished using in-house MATLAB-based graphical user interface named "IMRT3DCMP". For gel measurement dose grid from the TPS was extracted and compared with the measured dose grid of the gel. Gamma index analysis of film measurement for the tolerance criteria of 2%/2mm, 1%/1 mm showed more than 90% voxels pass rate. Gamma index analysis of 3D gel measurement data showed more than 90% voxels pass rate for different tolerance criteria of 2%/2 mm and 1%/1 mm. Overall both 2D and 3D measurement were in close agreement with the Pinnacle TPS calculated dose. The phantom designed is cost-effective and the results are promising, but further investigation is required to validate the phantom with other 3D conformal techniques for dosimetric purpose.

Role of Gamma Knife radiosurgery in trigeminal neuralgia - Its long term outcome and prediction using Artificial neural Network model.

PURPOSE: We aimed to study the long-term efficacy, prognostic factors and complications associated with the GKRS for trigeminal neuralgia. METHODS AND MATERIALS: Prospectively created database was analyzed for these patients. We created an Artificial neural Network - using Multilayer perceptron model in SPSS 23 by including all variables whose p value were<0.5 in univariate analysis. RESULTS: A total of 36 patients were included in the study. Patients pain free at 6 months were 25(69.44%), which reduced to 6(40 %) at 6 years. Median time to pain relief was 18.5 days. Only 5(13.88%) of them developed new onset or worsened numbness post Gamma Knife radio surgery. Median radiation dose was 80 Gy (prescribed at 100 percent isodose line). 1st GKT (P value < 0.05) and post GKT numbness (P value < 0.05) were the only factors favouring good outcome. Prior history of MVD was associated with poor pain relief post GKRS although p value was not significant (p = 0.136). ANN model could predict with 90.0 percent accuracy the favourable or unfavourable response on 11 Tested cases. In ANN model, a greater number of Pre GKT medications, previous MVD history, V2 dermatome involvement and negative history of post GKT numbness were negative prognostic factors. CONCLUSIONS: Lesser number of pre GKRS drugs used, involvement of V1 dermatome, post GKT numbness are favourable prognostic factors. Also, history of failed MVD for trigeminal neuralgia is associated with poor outcome. Repeat GKRS failed to show improvement in BNI grades.

Human immunodeficiency virus type-1 p24 sequence from an Indian strain: expression in Escherichia coli and implications in diagnostics.

A 637-bp fragment, corresponding to the p24 human immunodeficiency virus (HIV) core protein from the gag ORF, was PCR amplified from DNA isolated from peripheral blood mononuclear lymphocytes (PBML) of an asymptomatic HIV-1 seropositive human subject from Bombay and cloned into PCRScript SK(+). The nucleotide sequence revealed highest homology (98.6%) with the consensus sequence of the HIV-1 B subtype. The 637-bp KpnI-HindIII fragment was cloned downstream from a His6 tag in the pQE30 vector under the control of phage T5 promoter leading to production of a 6XHis-p24 fusion protein in Escherichia coli. It showed an approx. 24-kDa band by SDS-PAGE. The recombinant p24 reacted with serum samples from HIV-infected subjects when tested by Western blot and ELISA.

Identification of a novel surface mutant of hepatitis B virus in a seronegative chronic liver disease patient.

Hepatitis B virus (HBV) with mutations in the envelope proteins can emerge during natural infections, vaccinations or interferon therapy and appears occasionally to escape virus elimination or detection. The implications of such mutations at the molecular level are often obscure. We report the identification of a new surface mutant of HBV. This mutant was identified, and isolated from a chronic liver disease patient, negative for HBsAg as well as other serological markers but positive for HBV DNA. Several mutations were observed in the surface antigen gene out of which a Thr118-Ala118 change was predicted to have a destabilizing effect on the structural integrity of the 'a' determinant and also alter the antigenicity profile of the mutant HBsAg. Besides a RNA hairpin loop was predicted for the transcript generated by the small surface protein of this mutant, which could have an inhibitory effect at the translational level. These observations thus indicate that mutations in the surface gene could lead to a considerable decrease or complete absence of properly folded surface antigen which in turn could explain the absence of reactive HBsAg in the serum of the patient.

Clinico-immunological aspects of vernal catarrh in hilly terrains of Himachal Pradesh.

Very few immunological studies in vernal catarrh have been conducted in India and abroad, but none in Himachal Pradesh in spite of its high incidence in the State. In the present study 25 patients of vernal catarrh residing at a height ranging between 1000 to 2500 meters above mean sea level have been evaluated. Their immunological status of serum and tears after detailed clinical assessment was studied by single radial immunodiffusion technique of Mancini et al. The values of serum IgA and IgM were significantly higher in patients than in controls. The serum IgE level had no significant difference. The IgG was significantly lower in patients with vernal catarrh. The values of tear IgM, IgE and IgA in these patients were significantly higher than in controls. However, in no case or control group C3C and C4 were detected in tears. The limbal type of vernal catarrh was found to be the most common in this part of the country. No mixed case was seen. Derangement of the immune system in the pathogenesis of vernal catarrh is suggested.

SU-E-T-398: Verification of Gamma Knife EXtend System Based Fractionated Treatment Planning Using EBT2 Film.

PURPOSE: To present EBT2 film verification of treatment planning with the eXtend System, a relocatable frame system for multiple-fraction or serial multiple-session radiosurgery. METHODS: A human head shaped phantom simulated the verification process for fractionated Gamma Knife (GK) treatment. Phantom preparation for eXtend Frame based treatment planning involved creating a dental impression, fitting the phantom to the frame system, acquiring a stereotactic computed tomography (CT) scan. A CT scan (Siemens, Emotion6) of the phantom was obtained with following parameters: Tube Voltage - 110 kV, Tube Current - 280 mA, pixel size - 0.5 mm × 0.5 mm and 1 mm slice thickness. A treatment plan with two 8 mm collimator shots and three sector blocking in each shot was made. Dose prescription of 4.0 Gy at 100% was delivered for the first fraction out of the two fractions planned. Gafchromic EBT2 film (ISP Wayne, NJ) was used as 2D verification dosimeter in this process. Films were cut and placed inside the film insert of the phantom for treatment dose delivery. Meanwhile a set of films from the same batch were exposed from 0 Gy to 12 Gy doses for calibration purpose. EPSON (Expression 10000XL) scanner was used for scanning the exposed films in transparency mode. Scanned films were analyzed with in-house made Matlab codes. RESULTS: Gamma index analysis of film measurement in comparison with TPS calculated dose resulted in high pass rates >90% for different tolerance criteria of 2%/2mm, 1%/1mm, and 0.5%/0.5mm. The isodose overlay and linear dose profiles of film measured and computed dose distribution on sagittal and coronal plane was in close agreement. CONCLUSIONS: Through this study we propose a treatment verification QA method for eXtend frame based fractionated Gamma Knife radiosurgery using EBT2 film. Acknowledgement: Authors acknowledge the help of Andre Micke, ISP for sharing his expertise on EBT2 film.

SU-E-T-268: Evaluation of Photoneutron Contamination in Elekta Synergy-S High-Energy Linear Accelerator and Indigenous Novel Solution: The AIIMS Experience.

PURPOSE: The photoneutron contamination problem was encountered due to laminated barrier wall and short maze. The purpose of this study was to report our experience in evaluating the photoneutron contamination during radiation safety survey and solution. METHODS: The photoneutron contamination measurement was carried out in Elekta Synergy-S high-energylinear accelerator for 15MV beam. A NE Neutron survey meter and for photon, Victoreen and RADOS survey meters were used. The laminated barrier wall composed of 37cm steel with 30cm concrete both side and short maze length of 5 meter. During safety survey, higher photoneutron levels for 15MV X-rays at treatment room door found. The effect of photoneutron contamination as function of neutron shielding materials of wood, polyethylene and boron and thickness, distance, locations and directions to the control console at distance upto 7 meter were investigated for 4 gantry angles at locations of treatment room entry doors namely door1(A), door2(B), console(C), conduit (D) and above-ceiling(G) for 15MV. RESULTS: The initial safety survey showed that neutron level of 47mR/h and photon leakage of 3.2mR/hr at the treatment entry room door1. The neutron values could bring down to the level of acceptance at the treatment entry door2, but the photon values are not acceptable. Therefore, 30cm concrete wall block was made at the location of door2 and another bend was taken. Finally, treatment entrance room door was made using 3cm polyethylene neutron shielding materials in order to achieve the both neutron contamination and photon leakage within the acceptable levels. CONCLUSIONS: The neutron sliding-door is operated manually in finger-push by technologist for day-to-day usage. This simple solution is cost effective and increases the patient throughput. This study underlines that one needs to take appropriate safety measures prior to facility design whenever the space constraints situations arises for high energy linear accelerator.

Impact of different breathing conditions on the dose to surrounding normal structures in tangential field breast radiotherapy.

Cardiac toxicity is an important concern in tangential field breast radiotherapy. In this study, the impact of three different breathing conditions on the dose to surrounding normal structures such as heart, ipsilateral lung, liver and contralateral breast has been assessed. Thirteen patients with early breast cancer who underwent conservative surgery (nine left-sided and four right-sided breast cancer patients) were selected in this study. Spiral CT scans were performed for all the three breathing conditions, viz., deep inspiration breath-hold (DIBH), normal breathing phase (NB) and deep expiration breath-hold (DEBH). Conventional tangential fields were placed on the 3D-CT dataset, and the parameters such as V30 (volume covered by dose >30 Gy) for heart, V20 (volume covered by dose >20 Gy) for ipsilateral lung and V(50) (volume receiving >50% of the prescription dose) for heart and liver were studied. The average reduction in cardiac dose due to DIBH was 64% (range: 26.5-100%) and 74% (range: 37-100%) as compared to NB and DEBH respectively. For right breast cancer, DIBH resulted in excellent liver sparing. Our results indicate that in patients with breast cancer, delivering radiation in deep inspiration breath-hold condition can considerably reduce the dose to the surrounding normal structures, particularly heart and liver.

Sequence of complementary deoxyribonucleic acid encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP1 and its high-level expression in Escherichia coli.

Zona pellucida (ZP) glycoproteins have been proposed as candidate antigens for immunocontraception. Studies on this potential use can be facilitated by the availability of recombinant proteins. A cDNA lambda gt11 library was constructed using poly(A)+ mRNA isolated from bonnet monkey (Macaca radiata) ovaries and was screened for bonnet monkey ZP1 using a 404-basepair (bp) human ZP1 fragment (nucleotides 818-1221) as probe. Bonnet monkey ZP1 cDNA comprises 1617 nucleotides and encodes a polypeptide of 539 amino acid residues that share 92.0% identity with human ZP1. The major difference between bonnet monkey ZP1 and human ZP1 is the deletion of a 28-amino acid domain (amino acid residues 100-127 corresponding to human ZP1). An internal fragment (1317 bp) of bonnet monkey ZP1, excluding the N-terminus signal sequence and the C-terminus transmembrane-like domain, was amplified by polymerase chain reaction. The amplified Sac I and Kpn I restricted fragment was cloned in a frame downstream of the T5 promoter under the lac operator control for expression in the pQE-30 vector. Recombinant ZP1 (r-ZP1) was expressed as a polyhistidine fusion protein in Escherichia coli strains SG13009[pREP4] and ompT and Ion protease-deficient BL21 (plysS). SDS-PAGE analysis and immunoblotting with a murine monoclonal antibody, MA-410 (raised against porcine ZP3alpha--a homologue of bonnet monkey ZP1--and cross-reactive with bonnet monkey zona pellucida), revealed major bands of 51 and 40 kDa besides truncated fragments. Optimum expression of r-ZP1 was observed at 0.5 mM isopropyl beta-D-thiogalactopyranoside (IPTG). Immunization of male rabbits with r-ZP1 purified on nickel-nitrilotriacetic acid (NTA) resin under denaturing conditions and of female rabbits with r-ZP1 conjugated with diphtheria toxoid-generated antibodies reactive with r-ZP1 in ELISA. Moreover, immune sera, when tested by indirect immunofluorescence on bonnet monkey ovarian sections, showed positive fluorescence with zona pellucida. The information on the sequence of bonnet monkey ZP1 and the availability of the recombinant protein will help toward better understanding and evaluation of the contraceptive potential of homologous immunization in a nonhuman primate model.