ABO blood group discrepancies in blood donor and patient samples at a tertiary care oncology centre: analysis and serological resolution.

INTRODUCTION: In serological testing, determination of ABO grouping requires both antigen typing for A and B antigens and screening of serum or plasma for A and B antibodies. Lack of corroboration between the results of the cell and serum groupings identifies a discrepancy. Analysis of ABO blood group discrepancies was performed to determine the incidence of these discrepancies among healthy blood donors and oncology patients. MATERIALS AND METHODS: ABO discrepancies found during testing of blood samples from blood donors and patients in an oncology centre in the period from January 2015 to December 2018 were analysed. ABO blood grouping was performed using the column agglutination test. Detailed serological workups were carried out to resolve discrepancies. RESULTS: During the study period, a comprehensive analysis was conducted on a large dataset comprising 76,604 blood donor samples and 134,964 patient samples. Of these samples, 117 ABO discrepancies were identified with 13 occurring in blood donor samples and 104 in patient samples. The results demonstrated discrepancies caused by weakened/missing antibodies, weakened/missing antigens, panagglutination and miscellaneous factors in the blood donor samples, with percentages of 0%, 38%, 8%, and 31%, respectively. In patient samples, the percentages were 24%, 27%, 26%, and 15%, respectively. CONCLUSION: Weakened/missing antigen discrepancies were the prevalent type in both blood donor and patient samples. For accurate blood group reporting and management of transfusion needs of patients, a complete serological workup is vital to resolve any blood group discrepancies.

Navigating the neutropenic abyss with granulocyte transfusions: Retrospective single-center analysis of effectiveness and safety in India.

BACKGROUND: Hemato-oncologic patients receiving intensive chemotherapy may develop severe neutropenia and serious bacterial and/or fungal infections. Granulocyte transfusions (GTs) may be beneficial as a bridging therapy in hemato-oncologic patients with febrile neutropenia. AIM: To evaluate the clinical effectiveness of GTs in hemato-oncologic patients with febrile neutropenia. MATERIALS AND METHODS: This retrospective study evaluated the effectiveness of 150 GTs in 88 hemato-oncologic patients. Donors were mobilized with granulocyte colony-stimulating factors and dexamethasone. Patients' hematological parameters (pre- and post-GT) and safety and effectiveness of GTs were analyzed. RESULTS: The safety and effectiveness of GTs were assessed in the patients with various underlying conditions, including 78% with acute myeloid leukemia. In total, 150 GTs were administered, mostly during the chemotherapy induction phase. The GTs were well-tolerated by the patients, and a significant increment in white blood cell count and absolute neutrophil count (ANC) was noticed in 95% of patients after the transfusion. The granulocyte dose was positively correlated with ANC after the transfusion. The average time to neutrophil recovery from the last day of GT was 6.7 days, and the 30-day survival rate was 77%. The donors were all men, and a significant increase in WBC count was observed post-mobilization. The median granulocyte yield was 2.28 × 1010 /unit. All granulocyte products were crossmatched and irradiated before the transfusion. CONCLUSION: GTs can be a useful adjunctive treatment for febrile neutropenia in hemato-oncologic patients with multidrug-resistant sepsis. However, additional studies are required for confirming their effectiveness and establishing guidelines for their use.

Does granulocyte transfusion help pediatric cancer patients with sepsis?

Sepsis-related mortality continues to be a major concern while treating pediatric cancer patients, more so with the rise in the incidence of multidrug-resistant organisms (MDRO). In this retrospective study conducted between January 2021 and December 2022 at a tertiary cancer center in India, granulocyte transfusion was offered in addition to standard antimicrobial therapy to 64 children with hematolymphoid malignancy who developed 75 episodes of severe sepsis following intensive chemotherapy. Forty-four (83%) of 53 blood culture proven sepsis was caused by MDROs. Thirty-seven (70%) patients with blood culture proven sepsis cleared the organism after granulocyte transfusion. Thirty-day mortality was 25% for the entire study cohort and 32% for patients with MDRO sepsis.

The Case for Investing in Psychosocial Care and Rights of Children with Chronic Medical Conditions in India.

This narrative review paper aims to build a common understanding of the vulnerabilities of children with chronic medical conditions who face exceptional challenges due to nature of the illness, deformity, injury, and traumatic healthcare encounters which could potentially lead to long-term psychological effects. The presence of chronic medical diagnoses in children and the subsequent impacts including social stigma, as well as their age and developmental level, may amplify adjustment challenges in parenting considerations, school, peer relations, career, and future relationships. Children may be traumatized by unfamiliar and painful healthcare experiences. Hospitalization can lead to increased feelings of isolation, fear, and self-doubt when children do not receive emotionally safe psychological support necessary to minimize the accompanying stress and anxiety. School experiences and parenting children with chronic illnesses have additional intersecting socialization characteristics. This paper highlights a vision for furthering the groundwork within Indian pediatric settings to promote emotional safety and psychosocial care. Principles from the United Nations Convention on the Rights of the Child and the World Health Organization's definition of health create an impetus for giving a voice to children with chronic medical conditions. While there are sporadic psychosocial services for children with chronic needs, they lack consistency, and this indicates a need and an opportunity for developing a unique career for human development and psychology professionals to address these concerns. As exemplars, two psychosocial care programs in India that help foster resilience in these children are highlighted. Although limited, extant research regarding the experiences of children with chronic medical conditions in India is discussed, and implications for future research and academic initiatives are interwoven within this paper.

Serial monitoring of ionised calcium levels in plateletpheresis donors: A study from tertiary care oncology centre in India.

BACKGROUND: Plateletpheresis is a safe procedure, and the most common reaction is hypocalcemia which is transient and self-limiting, but it can have an impact on donor experience and donor return rate. AIM: To serially monitor the ionized calcium levels of the plateletpheresis donors and to correlate with symptoms of hypocalcemia if any. METHODS: It was a prospective observational study in 126 healthy voluntary donors eligible for plateletpheresis as per the Departmental SOP and after taking written informed consent. Procedures were conducted on continuous flow centrifugation (CFC) and intermittent flow centrifugation (IFC) cell separators. Donor blood samples were collected in pre-heparinized syringes at different intervals to measure ionized calcium levels (iCa++) by venous blood gas analysis (Cobas 221). RESULTS: There was a continuous and gradual decrease in iCa++ from start to 30-45 minutes during the procedure; while the levels showed a gradual increase at end of the procedure and reached near the baseline values after 15-30 min of completion of the procedure. The change in iCa++ was statistically significant at 30 min and 45 min (p < 0.05), which was correlated with symptoms of hypocalcemia observed in 32.5 % (41/126) of the donors. Females experienced more symptoms of hypocalcemia as compared to males (p < 0.01). Donors who underwent plateletpheresis on the IFC machine experienced more symptoms of hypocalcemia as compared to the CFC machine (p < 0.05). CONCLUSION: For donors with persistent symptoms of hypocalcemia which are unrelieved by procedural modifications (reducing blood return rate, citrate infusion rate, etc.) measurement of iCa++ and administration of oral calcium tablets may be considered.

Comparative evaluation of adhesion of glass fiber post to the root dentin after surface treatment of post and using universal bonding agent: An in vitro study.

Objective: The purpose of the study is the evaluation and comparison of surface roughness and bond strength of glass fiber post (GFP) after different types of surface treatment and the application of a universal bonding agent. Materials and Methods: Forty GFPs were divided into four groups based on surface treatment: Group I, silane coupling agent application for 60 s; Group II, air abrasion with 30 µm silicon dioxide powder particles in 2.5 bar pressure from 2 cm distance for 10 s, followed by silane coupling agent application; Group III, 9% hydrofluoric acid application for 10 s, rinsed and air-dried, followed by silane coupling agent application; and Group IV, silane coupling agent application, followed by universal bonding agent application. Surface roughness evaluation is done by a profilometer. All posts were cemented in the root of the maxillary central incisor with resin cement. After that, root was placed in an acrylic mold, and the external end part of the post was mounted on another acrylic mold. Pull-out bond strength was measured by a universal testing machine. Results: Highest surface roughness and bond strength values were found in Group II. Conclusion: Pretreatment of GFP increases the surface roughness of post as well as bond strength of post to root dentin. There is a correlation between surface roughness and bond strength. However, the use of only universal bonding agent also showed comparable pull-out bond strength of GFP, which means only use of universal bonding agent also a new alternative as pretreatment of GFP and helps in increase in bond strength.

Applying visual mapping techniques to promote learning in community-based medical education activities.

BACKGROUND: Teaching and learning Community-Based Medical Education (CBME) requires the active engagement of students in various activities to cover planned curricular content. CBME being multifaceted involves careful application and formation of links when attending to community health problems and public health issues. Students often depend on factual recall rather than 'engaging in', to counteract the broad and comprehensive nature of CBME. This study was conducted to assess the effectiveness of Visual mapping techniques as a learning tool in a CBME program for the subject Community Medicine and thereby assist medical undergraduate students in overcoming identified learning challenges. METHODOLOGY: An interventional study was conducted where medical undergraduates were randomly assigned to two different groups (each group = 30). After sensitization, a broad theme was taught to both the groups as a part of community-based teaching (CBT), each week for a month. The students in the intervention group were given the assignment to draw visual maps using the mind mapping & concept mapping techniques, after each CBT session, while the control group had Question-Answer session with built-in discussion (Conventional method) by an equally qualified, experienced faculty with no mapping assignments. A surprise written examination was conducted on the topics taught, and scores of both the groups were compared. Feedback was obtained from the intervention group. RESULTS: Mean score of the examination by the intervention group (29.85 ± 3.22) was significantly higher than and that of the control group (23.06 ± 4.09) (t = 7.14 and p < 0.05). The students shared that the assignment of drawing mind and concept maps for topics taught helped in attempting questions of the written examination by facilitating easy recall of the information learned. It aided to frame the answers to descriptive questions in a structured way with the use of keywords. However, identifying the concepts and establishing relationship between them was slightly challenging. CONCLUSION: 'Visual mapping' in the form of Mind and Concept mapping was found to be an effective learning tool for multifaceted CBME especially in promoting meaningful learning and facilitating rational thinking by the medical undergraduates.

EpiMix Based Novel Vaccine Candidate for Shigella: Evidence of Prophylactic Immunity in Balb/c Mice.

Multidrug resistant Shigella is one of the leading causes of mortality in children and infants. Availability of vaccine could prevent the Shigella infection and reduce the mortality. Conventional approaches of vaccine development against shigellosis have not resulted in desirable vaccine. As shigellosis may be caused by multiple strains and serotypes, there is a need to develop a multivalent vaccine, capable of providing protection against multiple Shigella strains. To develop broad spectrum vaccine, we had previously derived a pool of conserved epitopes against Shigella by using multiple immunoinformatic tools. In this study, the identified conserved epitopes derived from the Outer Membrane Proteins A and C of Shigella were chemically synthesized, and the EpiMix made up of 5 epitopes coupled to a carrier protein, ovalbumin was developed and validated for its immunogenicity. The intramuscular immunization with EpiMix in Balb/c mice led to increase in EpiMix specific serum IgG, and significant increase in fecal IgA as well as in IL-4, IL-2and IFN-γ levels. Further, the EpiMix immunized mice showed protection when challenged against S. flexneri ATCC 12022 using the intraperitoneal route. Moreover, the analysis of cytokine profile and IFN-γ/IL4 ratio in post Shigella challenge immunized mice suggested the high levels of IFN-γ levels and possible dominance of Th1 response, playing pivotal role in the elimination of Shigella. Collectively, the results demonstrate the immunogenic potential and protective efficacy of the EpiMix in the murine shigellosis model. However, the detailed study and further optimisation of epitopes would substantiate the prospective use of EpiMix as a prophylactic candidate for vaccination.

Evaluation of debris and smear layer formation using three different NI-TI rotary instrument systems: An in vitro scanning electron microscope study.

Aim: The aim of the study is to evaluate of debris and smear layer formation after using rotary ProTaper Universal, Twisted File, and XP Endo file systems under scanning electron microscope. Materials and Methods: Forty freshly extracted mandibular second premolar teeth were taken to decoronate at the cementoenamel junction to make the remaining root length 15 mm. Specimens were divided into four groups of 10 teeth each, Group I (control) - no instrumentation. Group II - ProTaper Universal rotary file (F2), Group III - twisted file (ISO size 0.25 and 6% taper), Group IV - XP Endo file (ISO size 0.25). During instrumentation, 5 ml normal saline was used as irrigating agent. Grooves parallel to the longitudinal axis of the root were made on the mesial and distal surface of each specimen to split it into two halves and examined under scanning electron microscope at ×1500 and ×5000 magnification. Photomicrographs were taken to evaluate debris and smear layer. Evaluation of photomicrographs was done using a score index. Results: One-way analysis of variance (ANOVA) was used to compare more than one means at a time. Tukey's critical difference followed by ANOVA was used to compare the mean values pair wise. P <0.05 was considered to be statistically significant. Among all the file systems, Group II showed maximum amount of debris (3.50 ± 1.109) followed by Group III (2.83 ± 1.238) and least amount was showed by Group IV (2.65 ± 1.122) at all levels (cervical, middle, and apical third). Among all the experimental groups, Group II showed maximum amount of smear layer (2.75 ± 1.149) followed by Group III (2.40 ± 0.982) and least amount of smear layer shown by Group IV (2.10 ± 0.841) at all levels (cervical, middle and apical third), the result was statistically significant (P < 0.05). Conclusions: At all the levels (cervical third, middle third, and apical third), among all the experimental groups, highest amount of debris and smear layer was formed by ProTaper Universal rotary file followed by Twisted file and least amount showed by XP Endo file system. In all the levels, control group showed highest amount of debris but least amount of smear layer.

Unexpected high platelet counts in a potential platelet donor: A clue to essential thrombocythemia.

Essential thrombocythemia (ET) (primary thrombocythemia) is a nonreactive, chronic myeloproliferative clonal disorder in which sustained megakaryocyte proliferation leads to an increase in the number of circulating platelets. It is silent disorder and is diagnosed as high platelet counts with or without associated symptoms or as an accidental finding. We, hereby, report a case of ET as an incidental finding during screening of a potential platelet donor. Donors with high platelet count should be investigated properly to rule out ET. The role of transfusion medicine specialist is not only to recruit donors but also to identify such donors during screening and counsel them for detailed investigation and proper management.

Practices and Perceptions of Nurses Regarding Child Visitation in Adult Intensive Care Units.

BACKGROUND: Provision of developmentally appropriate support for child visitors in adult intensive care units (ICUs) would benefit patients and young visitors. Research on best practices for child visitation in adult ICUs is limited. OBJECTIVES: To explore the perceptions and practices of nurses working in adult ICUs in the United States regarding child visitation and the role of child life specialists in this setting. METHODS: Data were collected from 446 adult ICU nurses via a cross-sectional survey. The survey explored perceptions and practices regarding child visitation, access to child-friendly resources, and the feasibility of having a child life specialist in adult ICUs. RESULTS: Several participants (303, 67.9%) felt that children were at risk for psychological trauma from visiting an adult ICU. Some participants (122, 27.4%) reported that their ICUs did not have policies for child visitation. Logistic regression showed that nurses with a master's degree were 1.8 times (P < .05) more likely to believe that young children (0-5 years) should visit. Nurses (105 of 197, 53.3%) were more likely to allow young children to visit if the patient was the child's parent or if the patient was dying. Child-friendly resources were not routinely available. Nurses expressed that adult ICUs could benefit from child life specialists facilitating child visitation. CONCLUSIONS: Nurses were inconsistently open to child visitation. Exceptions for older children (> 6 years), children whose parent was the patient, patients' illness severity, and end of life allowed more child visitation. Ways to facilitate child-friendly visitation in adult ICUs are discussed.

Management of transfusion needs in a case of immunizing anti-M antibody in a patient with acute myeloid leukemia.

Anti-M is a relatively common "naturally occurring" antibody. Unexpected alloantibodies in patient's serum other than ABO isoagglutinins (e.g., anti-M) may cause a discrepancy in the reverse grouping. As long as anti-M does not react at 37°C, it is clinically insignificant for transfusion. However, we found this antibody to be of "immunizing" type which was reactive at 37°C and AHG phase and showing problems in blood grouping and crossmatch. This antibody had both IgM and IgG components. When "M" antibodies active at 37°C are encountered, antigen-negative or red cells that are compatible with an indirect antiglobulin test should be provided.

Immunoinformatic Identification of Potential Epitopes.

Immunoinformatic plays a pivotal role in vaccine design and development. While traditional methods are exclusively depended on immunological experiments, they are less effective, relatively expensive, and time-consuming. However, recent advances in the field of immunoinformatics have provided innovative tools for the rational design of vaccine candidates. This approach allows the selection of immunodominant regions from the sequence of whole genome of a pathogen. The identified immunodominant region could be used to develop potential vaccine candidates that can trigger protective immune responses in the host. At present, epitope-based vaccine is an attractive concept which has been successfully trailed to develop vaccines against a number of pathogens. In this chapter, we outline the methodology and workflow of how to deploy immunoinformatics tools in order to identify immunodominant epitopes using Shigella as a model organism. The immunodominant epitopes, derived from S. flexneri 2a using this workflow, were validated using in vivo model, indicating the robustness of the outlined workflow.

Comparative evaluation of different probiotic products on salivary Streptococcus mutans and Lactobacillus level in caries risk population.

INTRODUCTION: Dental caries is a multifactorial disease with the main causative organism being Streptococcus mutans and Lactobacillus spp. "Probiotics" are defined as living microorganisms, principally bacteria, which are safe for human consumption and when ingested in sufficient quantities, have beneficial effects on human health, beyond basic nutrition. These can be used to replace the pathogenic strains of bacteria with the nonpathogenic type in the oral cavity thus can help prevent dental caries. AIMS: To evaluate and compare the role of probiotic milk and lozenges on S. mutans and Lactobacillus spp. count on patients who are exposed to probiotics continuously for 7 days. MATERIALS AND METHODS: Sixty volunteers who were prone to caries were divided into three equal groups. Experimental groups were given probiotics daily for 7 days. In Group A, patients were given milk without any probiotics (control); in Group B, probiotic milk is given; and in Group C, probiotic lozenges are given. Saliva samples were assessed on the 1st day and after 7 days of intervention. The collected saliva samples were inoculated on the selected culture media and estimation was done by measuring the colony-forming unit. STATISTICAL ANALYSIS USED: Statistical analysis was performed using Student's paired t-test and multiple comparisons by Tukey's honest significant difference test. RESULTS: There was a significant reduction in salivary S. mutans level in both experimental groups after 7 days (P < 0.05). However, there was no significant difference in Lactobacillus count before and after the intervention. CONCLUSIONS: Probiotics have a role in reducing salivary S. mutans count. Probiotics lozenges showed greater efficacy in reducing salivary S. mutans count than probiotic milk.

Comparative evaluation of surface roughness and color stability of nanohybrid composite resin after periodic exposure to tea, coffee, and Coca-cola - An in vitro profilometric and image analysis study.

AIM: The aim of this study is to evaluate the surface roughness and color stability of a nanohybrid composite resin after exposure to tea, coffee, Coca-cola, and artificial saliva on the 7th, 14th, and 28th day. MATERIALS AND METHODS: Forty specimens of nanohybrid composite resin discs of dimension 6 mm × 2 mm were fabricated and were randomly divided into four equal groups based on beverages (Group 1: control group - artificial saliva, Group 2: Tea, Group 3: Coffee, and Group 4 : Coca-cola). Baseline readings of surface roughness (Ra) and color change (ΔE) were obtained by profilometer and digital image analysis technique in CIE l*a*b scale followed by the readings (ΔE) taken on the 7th, 14th, and 28th day of exposure to respective beverages. STATISTICAL ANALYSES: One-way analysis of variance (ANOVA) followed by post hoc Tukey's test and paired t-test along with Pearson's correlation with a P = 0.05. RESULTS: Overall, highest surface roughness was obtained in the samples exposed to Coca-cola followed by coffee and tea while the least surface roughness was obtained in the samples exposed to artificial saliva (control group) which was statistically significant (P < 0.05). Overall highest color change was obtained in tea followed by coffee and Coca-cola. The least color change was observed in the samples exposed to artificial saliva (control group) (P < 0.05). CONCLUSION: Surface roughness and color change were time dependent as both increased with time. Furthermore, greater the surface roughness, more was the color change observed in all the groups at all tested time intervals.

Oral immunization with LacVax® OmpA induces protective immune response against Shigella flexneri 2a ATCC 12022 in a murine model.

Shigellosis is an acute invasive disease of the lower intestine, which afflicts millions of people worldwide with an estimated one million fatalities per annum. Despite of extensive research during the last two decades, a vaccine against multi-drug resistant Shigella is not yet available in the market. To provide a safe, effective and broad-spectrum vaccine against Shigella, we explored food grade bacteria Lactococcus lactis (L. lactis) for the delivery of conserved antigenic protein; Outer membrane protein A (OmpA) to the mucosal sites for effective elicitation of systemic and mucosal immunity. We have previously confirmed the immunogenic potential of recombinant L. lactis expressing OmpA (LacVax® OmpA) in BALB/c mice. In the present study, we have characterized the humoral and cellular immune profile of LacVax® OmpA and assessed its protective efficacy using a newly developed human like murine shigellosis model. The significant increase in OmpA specific serum IgG, fecal sIgA and a Th1 dominant immune response (indicated by high INF-γ/IL-4 ratio) in LacVax® OmpA immunized mice revealed successful activation of humoral and cellular immunity. The LacVax® OmpA immunized animals were also protected from human-like shigellosis when challenged with S. flexneri 2a ATCC 12022. The antigen specific serum IgG, fecal sIgA, INF-γ and IL-10 levels were found to be the significant correlates of protection. Collectively these results suggest that the LacVax® OmpA is a promising prophylactic candidate against shigellosis. However, the protective efficacy of LacVax® OmpA in the higher animals would further strengthen its future application in humans.

Microbubble-Mediated Enhanced Delivery of Curcumin to Cervical Cancer Cells.

The major bottleneck in the current chemotherapy treatment of cancer is the low bioavailability and high cytotoxicity. Targeted delivery of drug to the cancer cells can reduce the cytotoxicity and increase the bioavailability. In this context, microbubbles are currently being explored as drug-delivery vehicles to effectively deliver drug to the tumors or cancerous cells. Microbubbles when used along with ultrasound can enhance drug uptake and inhibit the growth of tumor cells. Several potential anticancer molecules exhibit poor water solubility, which limits their use in therapeutic applications. Such poorly water soluble molecules can be coadministered with microbubbles or encapsulated within or loaded on the microbubbles surface, to enhance the effectiveness of these molecules against cancer cells. Curcumin is one of such potential anticancer molecules obtained from the rhizome of herbal spice, turmeric. In this work, curcumin-loaded protein microbubbles were synthesized and examined for effective in vitro delivery of curcumin to HeLa cells. Microbubbles in the size range of 1-10 µm were produced using perfluorobutane as core gas and bovine serum albumin (BSA) as shell material and were loaded with curcumin. The amount of curcumin loaded on the microbubble surface was estimated using UV-vis spectroscopy, and the average curcumin loading was found to be ∼54 µM/108 microbubbles. Kinetics of in vitro curcumin release from microbubble surface was also estimated, where a 4-fold increase in the rate of curcumin release was obtained in the presence of ultrasound. Sonication and incubation of HeLa cells with curcumin-loaded BSA microbubbles enhanced the uptake of curcumin by ∼250 times. Further, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay confirmed ∼71% decrease in cell viability when HeLa cells were sonicated with curcumin-loaded microbubbles and incubated for 48 h.

In vivo delivery of pPERDBY to BALB/c mice by LacVax® DNA-I and comparison of elicited immune response with conventional immunization methods.

The non-invasive food grade Lactococcus lactis (L. lactis) represents a safe and attractive alternative to invasive pathogens for the delivery of plasmid DNA at mucosal sites. We have earlier shown the DNA delivery potential of r-L. lactis harboring DNA vaccine reporter plasmid; pPERDBY in vitro. In the present work, we examined in vivo delivery potential of food grade non-invasive r-L. lactis::pPERDBY (LacVax® DNA-I) in BALB/c mice. Moreover, using EGFP as a model antigen, we also characterized and compared the immune response elicited by LacVax® DNA-I with other conventional vaccination approaches using protein and naked DNA immunization. The presence of antigen-specific serum IgG and fecal secretory IgA (sIgA) antibodies demonstrated in vivo DNA delivery and immune elicitation potential of the developed LacVax® DNA-I. As compared with intramuscular injection, oral delivery of pPERDBY via L. lactis resulted in a significantly rapid increase in IgG and higher sIgA titers, indicating the immunogenic and immunostimulatory properties of the LacVax® DNA-I. The needle-free immunization with LacVax® DNA-I led to increased production of IL-4, an indicator of Th2 screwed response. To the best of our knowledge, this report for the first time outlines comparison of orally administered LacVax® DNA-I with other conventional vaccination approaches.

Comparative evaluation of three methods to measure working length - Manual tactile sensation, digital radiograph, and multidetector computed tomography: An in vitro study.

AIM: Compare the measurement of working length with three different methods manual tactile sensation, digital radiography and Mutidetector computed tomography(MDCT). MATERIALS AND METHOD: 40 human premolar extracted for orthodontic purpose were selected. Teeth were store in sodium chloride (0.9%) during the study. Access cavity was prepared and canal patency was seen with no 10 file in each tooth. Manually no 15 K file was inserted from access cavity until the tip was visible at the foramen, a silicon stopper was adjusted to the corresponding buccal cusp tip and the root canal length was measured. After that in each tooth with no-15 K file inserted from access cavity with stop at tip of buccal cusp until tip appear at foramen and x-ray was taken with digital radiograph (RVG,Satelac) and canal length was measured. After that all teeth are mounted in wax block,MDCT scan was done and in the scan images of teeth, root canal length is measured from buccal cusp tip to root end. After taking measurement of working length with all three methods and the data was stastically analyzed with One Way Analysis of variance (ANOVA) followed by Turkey's Test. RESULTS: ANOVA and turkeys test showed that there was no significant difference in the measurements by the three procedures (p>0.05). CONCLUSION: Working length measurement with MDCT scan and other two conventional methods does not show significant difference in measurement. Use of newer 3D imaging technique is useful in root canal treatment for measuring working length.

Dual recombinant Lactococcus lactis for enhanced delivery of DNA vaccine reporter plasmid pPERDBY.

Food grade Lactococcus lactis has been widely used as an antigen and DNA delivery vehicle. We have previously reported the use of non-invasive L. lactis to deliver the newly constructed immunostimulatory DNA vaccine reporter plasmid, pPERDBY. In the present report, construction of dual recombinant L. lactis expressing internalin A of Listeria monocytogenes and harboring pPERDBY (LL InlA + pPERDBY) to enhance the efficiency of delivery of DNA by L. lactis is outlined. After confirmation and validation of LL InlA + pPERDBY, its DNA delivery potential was compared with previously developed non-invasive r- L. lactis::pPERDBY. The use of invasive L. lactis resulted in around threefold increases in the number of enhanced green fluorescent protein-expressing Caco-2 cells. These findings reinforce the prospective application of invasive strain of L. lactis for delivery of DNA/RNA and antigens.