Comparison of noninvasive cardiac output monitoring by electrical cardiometry with transthoracic echocardiography in postoperative paediatric cardiac surgical patients - A prospective observational study.

Aim: The present study was conducted to validate cardiac output (CO) and cardiac index (CI) obtained from electrical cardiometry (EC) ICON ® with transthoracic echocardiography (TTE) in postoperative pediatric cardiac surgical patients. Materials and Methods: A prospective observational study was conducted in 25 pediatric patients with age < 10 years who underwent elective cardiac surgery. Data Analysis: BlandAltman plot was constructed for interchangeability and Polar plot was constructed to know trending ability. Results: A total of 250 datasets were analyzed. Spearman's correlation coefficient for CO between ICON ® and TTE showed good positive correlation (r = 0.850, 95% confidence interval 0.81 to 0.881, P <.0001). Moderate positive correlation was observed between ICON ® and TTE for CI (r = 0.60, 95% confidence interval 0.515 to 0.674, P <.0001). Linear regression equations for CO and CI between ICON ® and TTE were: y = 0.5230 + 0.8078 X (R2 = 0.6597, P <.001) and y = 1.8350 + 0.5869 X (R2 = 0.3985, P <.001) [y- ICON ®; X - TTE], respectively. BlandAltman plot for CO between ICON ® and TTE showed a bias of 0.3012 with limits of agreement (LOA) being -0.69 to 1.3 and for CI bias was 0.6939 with LOA-2.1 to 3.5. Polar plot analysis showed an angular bias of 8.1750, with radial LOA being -13.74° to 30.08° for CO and angular bias of 6.6931, with radial LOA being -15.69° to 29.07° for CI. Conclusion: ICON ® monitor-derived parameters are not interchangeable with the values derived from TTE. However, the ICON ® monitor demonstrated a good trending ability for both CO and CI.

Adult Onset Sacrococcygeal Teratoma.

Sacrococcygeal teratoma (SCT), one of the most common neoplastic tumors in newborns, is found very rarely in adults. These teratomas are germ cell tumours. Most of these tumors are benign and cystic in nature, with only 1-2% of them having a malignant transformation. Most of these tumors are benign and cystic in nature, with only 1-2% of them having malignant transformation. A higher incidence was seen in females. Typically, cystic teratomas are asymptomatic, and so the diagnosis was often made inadvertently during radiographic studies. The majority of treatment is complete surgical excision, and both open and laparoscopic procedures have been proven to be efficient. Histopathologic examination can confirm the diagnosis. We present this unusual instance of a 56-year-old female patient with a sacrococcygeal teratoma.

Stroke Lesion Segmentation in FLAIR MRI Datasets Using Customized Markov Random Fields.

Robust and reliable stroke lesion segmentation is a crucial step toward employing lesion volume as an independent endpoint for randomized trials. The aim of this work was to develop and evaluate a novel method to segment sub-acute ischemic stroke lesions from fluid-attenuated inversion recovery (FLAIR) magnetic resonance imaging (MRI) datasets. After preprocessing of the datasets, a Bayesian technique based on Gabor textures extracted from the FLAIR signal intensities is utilized to generate a first estimate of the lesion segmentation. Using this initial segmentation, a customized voxel-level Markov random field model based on intensity as well as Gabor texture features is employed to refine the stroke lesion segmentation. The proposed method was developed and evaluated based on 151 multi-center datasets from three different databases using a leave-one-patient-out validation approach. The comparison of the automatically segmented stroke lesions with manual ground truth segmentation revealed an average Dice coefficient of 0.582, which is in the upper range of previously presented lesion segmentation methods using multi-modal MRI datasets. Furthermore, the results obtained by the proposed technique are superior compared to the results obtained by two methods based on convolutional neural networks and three phase level-sets, respectively, which performed best in the ISLES 2015 challenge using multi-modal imaging datasets. The results of the quantitative evaluation suggest that the proposed method leads to robust lesion segmentation results using FLAIR MRI datasets only as a follow-up sequence.

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development.

Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

Recent Developments in Recombinant Protein-Based Dengue Vaccines.

Recombinant proteins are gaining enormous importance these days due to their wide application as biopharmaceutical products and proven safety record. Various recombinant proteins of therapeutic and prophylactic importance have been successfully produced in microbial and higher expression host systems. Since there is no specific antiviral therapy available against dengue, the prevention by vaccination is the mainstay in reducing the disease burden. Therefore, efficacious vaccines are needed to control the spread of dengue worldwide. Dengue is an emerging viral disease caused by any of dengue virus 1-4 serotypes that affects the human population around the globe. Dengue virus is a single stranded RNA virus encoding three structural proteins (capsid protein, pre-membrane protein, and envelope protein) and seven non-structural proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5). As the only licensed dengue vaccine (Dengvaxia) is unable to confer balanced protection against all the serotypes, therefore various approaches for development of dengue vaccines including tetravalent live attenuated, inactivated, plasmid DNA, virus-vectored, virus-like particles, and recombinant subunit vaccines are being explored. These candidates are at different stages of vaccine development and have their own merits and demerits. The promising subunit vaccines are mainly based on envelope or its domain and non-structural proteins of dengue virus. These proteins have been produced in different hosts and are being investigated for development of a successful dengue vaccine. Novel immunogens have been designed employing various strategies like protein engineering and fusion of antigen with various immunostimulatory motif to work as self-adjuvant. Moreover, recombinant proteins can be formulated with novel adjuvants to enhance the immunogenicity and thus conferring better protection to the vaccinees. With the advent of newer and safer host systems, these recombinant proteins can be produced in a cost effective manner at large scale for vaccine studies. In this review, we summarize recent developments in recombinant protein based dengue vaccines that could lead to a good number of efficacious vaccine candidates for future human use and ultimately alternative dengue vaccine candidates.

Development of nsP2 protease based cell free high throughput screening assay for evaluation of inhibitors against emerging Chikungunya virus.

Chikungunya virus has emerged as one of the most important global arboviral threats over the last decade. Inspite of large scale morbidity, with long lasting polyarthralgia, so far no licensed vaccine or antiviral is available. CHIKV nsP2 protease is crucial for processing of viral nonstructural polypeptide precursor to release enzymes required for viral replication, thus making it a promising drug target. In this study, high cell density cultivation (HCDC) of Escherichia coli in batch process was carried out to produce rCHIKV nsP2pro in a cost-effective manner. The purified nsP2pro and fluorogenic peptide substrate have been adapted for fluorescence resonance energy transfer (FRET) based high throughput screening (HTS) assay with Z' value and CV of 0.67 ± 0.054 and <10% respectively. We used this cell free HTS system to screen panel of metal ions and its conjugate which revealed zinc acetate as a potential candidate, which was further found to inhibit CHIKV in Vero cells. Scale-up process has not been previously reported for any of the arboviral nonstructural enzymes. The successful scale-up method for viral protease together with a HTS assay could lead to the development of industrial level large-scale screening platform for identification of protease inhibitors against emerging and re-emerging viruses.

Enhanced production and immunological characterization of recombinant West Nile virus envelope domain III protein.

West Nile virus (WNV) is an emerging mosquito-borne virus which is responsible for severe and fatal encephalitis in humans and for which there is no licensed vaccine or therapeutic available to prevent infection. The envelope domain III protein (EDIII) of WNV was over-expressed in Escherichia coli and purified using a two-step chromatography process which included immobilized metal affinity chromatography and ion exchange chromatography. E. coli cells were grown in a bioreactor to high density using batch and fed-batch cultivation. Wet biomass obtained after batch and fed-batch cultivation processes was 11.2 g and 84 g/L of culture respectively. Protein yield after affinity purification was 5.76 mg and 5.81 mg/g wet cell weight after batch and fed-batch processes respectively. The purified WNV EDIII elicited specific antibodies in rabbits, confirming its immunogenicity. Moreover, the antibodies were able to neutralize WNV in vitro. These results established that the refolded and purified WNV EDIII could be a potential vaccine candidate.

Working towards dengue as a vaccine-preventable disease: challenges and opportunities.

INTRODUCTION: Dengue is an emerging viral disease that affects the human population around the globe. Recent advancements in dengue virus research have opened new avenues for the development of vaccines against dengue. The development of a vaccine against dengue is a challenging task because any of the four serotypes of dengue viruses can cause disease. The development of a dengue vaccine aims to provide balanced protection against all the serotypes. Several dengue vaccine candidates are in the developmental stages such as inactivated, live attenuated, recombinant subunit, and plasmid DNA vaccines. Area covered: The authors provide an overview of the progress made in the development of much needed dengue vaccines. The authors include their expert opinion and their perspectives for future developments. Expert opinion: Human trials of a live attenuated tetravalent chimeric vaccine have clearly demonstrated its potential as a dengue vaccine. Other vaccine candidate molecules such as DENVax, a recombinant chimeric vaccine andTetraVax, are at different stages of development at this time. The authors believe that the novel strategies for testing and improving the immune response of vaccine candidates in humans will eventually lead to the development of a successful dengue vaccine in future.

Morphometric evaluation of the foramen magnum for sex determination: A study from Saudi Arabia.

The present study provides a database of various morphometric dimensions of the foramen magnum region in the Saudi population. The objective of this study was to evaluate various measurements of the foramen magnum region for sex determination in the Saudi population by using computed tomography (CT) images. The various radiological measurements of the foramen magnum region were measured in a total of 200 adult subjects of Saudi origin including 100 males and 100 females. Sexual dimorphism was observed in five parameters related to the foramen magnum, namely length of the right occipital condyle (LROC), length of the left occipital condyle (LLOC), width of the foramen magnum (WFM), area of the foramen magnum (AFM) and length of the foramen magnum (LFM). The accuracy to discriminate sex ranged from 65.5% to 62.5% when LROC, LLOC, WFM, AFM, and LFM were considered as individual parameters. When multiple parameters were combined to discriminate sex, the highest accuracy of 71% was achieved.

Evaluation of antibody response against recombinant domain III proteins of dengue virus type 1 and 2.

Dengue, a mosquito borne viral disease caused by dengue virus has emerged as a major health problem during the last few decades. The envelope domain III (DIII) protein of dengue virus is highly immunogenic and capable of inducing neutralizing antibodies against wild-type dengue virus. The envelope domain III protein is a potential subunit vaccine candidate as well as a diagnostic reagent for dengue. This report describes the high yield production and immunogenicity of recombinant DIII proteins of dengue virus type 1 and 2. The subunit DIII proteins were produced in Escherichia coli using batch and fed-batch fermentation process. Immobilized metal affinity chromatography was used to capture DIII proteins of dengue virus type 1 and 2. The purified proteins were refolded by diafiltration to achieve biologically active proteins. After fed-batch fermentation, the recombinant E. coli resulted in purified DIII proteins of about 10.06 mg and 47.70 mg per gram of dry cell weight for recombinant dengue virus type 1 and 2 respectively with more than 95% purity. Biological function of the purified DIII proteins were confirmed by their ability to generate DIII specific antibodies in mice. The DIII antigens in combination with adjuvant resulted antibody endpoint titers of 1:64,000 and 1:1,28,000 for recombinant dengue virus type 1 and 2 respectively. These findings establish that the DIII proteins in combination with adjuvant are immunogenic, which suggests that refolded and purified DIII proteins can be a potential vaccine candidates.

GSTM1 null polymorphism prevalence in tobacco users, oral leukoplakia and oral squamous cell carcinoma patients in South Indian population: A polymerase chain reaction study.

CONTEXT: Tobacco abuse is a well-known risk factor for potentially malignant disorders as well as oral squamous cell carcinoma. Factors that influence tobacco-exposed individuals developing a malignancy may include the combination of total tobacco exposure and genetic susceptibility. AIM: This study was undertaken to determine the prevalence of the glutathione S-transferase M1 (GSTM1) null polymorphism in oral leukoplakia and oral squamous cell carcinoma patients in South Indian population. SETTINGS AND DESIGN: This case-control study was conducted in hospital setting on South Indian population. MATERIALS AND METHODS: About 280 subjects with history of tobacco use, oral leukoplakia, oral squamous cell carcinoma were included in this study. Three milliliter of blood was collected and transported under cold cycle and taken for evaluation of GSTM1 null polymorphism using multiplex polymerase chain reaction. RESULTS AND DISCUSSION: On comparing the prevalence of GSTM1 null polymorphism among the group with subjects with habits and no oral lesions, oral leukoplakia, and oral squamous cell carcinoma, it was observed that there was a statistically significant association between GSTM1 null polymorphism and the different groups (P < 0.01). CONCLUSION: The lack of GSTM1 activity would make the oral tissues more susceptible to action of tobacco carcinogens and to the development of a high-grade level of dysplasia in oral leukoplakia and thereby increases the susceptibility of lesion to undergo malignant changes.

Feasibility of measuring superior mesenteric artery blood flow during cardiac surgery under hypothermic cardiopulmonary bypass using transesophageal echocardiography: An observational study.

BACKGROUND: Abdominal complications being rare but results in high mortality, commonly due to splanchnic organ hypoperfusion during the perioperative period of cardiac surgery. There are no feasible methods to monitor intraoperative superior mesenteric artery blood flow (SMABF). Hence, the aim of this study was to evaluate the feasibility and to measure SMABF using transesophageal echocardiography (TEE) during cardiac surgery under hypothermic cardiopulmonary bypass (CPB). METHODOLOGY: Thirty-five patients undergoing elective cardiac surgery under CPB were enrolled. Heart rate, mean arterial pressure (MAP), cardiac output (CO), SMABF, superior mesenteric artery (SMA) diameter, superior mesentric artery blood flow over cardiac output (SMA/CO) ratio and arterial blood lactates were recorded at three time intervals. T0: before sternotomy, T1: 30 min after initiation of CPB and T2: after sternal closure. RESULTS: SMA was demonstrated in 32 patients. SMABF, SMA diameter, SMA/CO, MAP and CO-decreased significantly (P < 0.0001) between T0 and T1, increased significantly ( P ≤ 0.001) between T1 and T2 and no significant change (P > 0.05) between T0 and T2. Lactates increased progressively from T0 to T2. CONCLUSION: Study shows that there is decrease in SMABF during CPB and returns to baseline after CPB. Hence, it is feasible to measure SMABF using TEE in patients undergoing cardiac surgery under hypothermic CPB. TEE can be a promising tool in detecting and preventing splanchnic hypoperfusion during perioperative period.

Estimation of gestational age from gall-bladder length.

Establishing a precise duration of gestation is vital in situations such as infanticide and criminal abortions. The present study attempted to estimate the gestational age of the foetus from gall-bladder length. Foetuses of various gestational age groups were dissected, and the length of the gall bladder was measured. The results were analysed, and a substantial degree of correlation was statistically confirmed. This novel method is helpful when the foetus is fragmented, putrefied or eviscerated, where this method can be used as an additional parameter to improve the accuracy of foetal age estimation.

Evaluation of the mastoid triangle for determining sexual dimorphism: A Saudi population based study.

Demographic assessment of skeletal remains in forensic investigations includes identification of sex. The present study aimed to develop population-specific, sex-discriminating anthropometric standards for the mastoid triangle of a documented Saudi population using computed tomographic (CT) images of the lateral aspect of the skull. The present study was performed on 206 CT images of a documented Saudi population of known sex and age. The clinical CT images of subjects visiting the Department of Radiology, Dammam Medical Complex, Dammam, Saudi Arabia (KSA) were evaluated to know the validity of the metric assessment of the mastoid triangle for identification of sex in a Saudi population. The distance between asterion to porion (AP), asterion to mastoidale (AM), porion to mastoidale (PM) were measured and the area of the mastoid triangle (AMT) was calculated using these measurements. Discriminant function procedure was used to analyze the data for sexual dimorphism. In conclusion, the results of the present study indicate that all the 3 sides of the mastoid triangle and AMT were sexually dimorphic in the sampled Saudi population with PM being the best individual parameter in discriminating sex with an accuracy of 69.4%. Whereas, all the parameters combined showed the highest accuracy (71.4%).

Demirjian's method in the estimation of age: A study on human third molars.

AIM: The primary aim of the following study is to estimate the chronological age based on the stages of third molar development following the eight stages (A to H) method of Demirjian et al. (along with two modifications-Orhan) and secondary aim is to compare third molar development with sex and age. MATERIALS AND METHODS: The sample consisted of 115 orthopantomograms from South Indian subjects with known chronological age and gender. Multiple regression analysis was performed with chronological age as the dependable variable and third molar root development as independent variable. All the statistical analysis was performed using the SPSS 11.0 package (IBM ® Corporation). RESULTS: Statistically no significant differences were found in third molar development between males and females. Depending on the available number of wisdom teeth in an individual, R (2) varied for males from 0.21 to 0.48 and for females from 0.16 to 0.38. New equations were derived for estimating the chronological age. CONCLUSION: The chronological age of a South Indian individual between 14 and 22 years may be estimated based on the regression formulae. However, additional studies with a larger study population must be conducted to meet the need for population-based information on third molar development.

Prevalence of glutathione S-transferase M1 null polymorphism in tobacco users, oral leukoplakia and oral squamous cell carcinoma patients in South Indian population: A polymerase chain reaction study.

CONTEXT: Tobacco abuse is a well-known risk factor for potentially malignant disorders as well as oral squamous cell carcinoma (SCC). Factors that influence tobacco-exposed individuals developing a malignancy may include a combination of total tobacco exposure and genetic susceptibility. AIM: This study was undertaken to determine the prevalence of the glutathione S-transferase M1 (GSTM1) null polymorphism in oral leukoplakia and oral SCC patients in South Indian population. SETTINGS AND DESIGN: This case-control study was conducted in hospital setting on South Indian population. MATERIALS AND METHODS: Totally, 280 subjects with a history of tobacco use, oral leukoplakia, oral SCC were included in this study. Three milliliter of blood was collected and transported under cold cycle and taken for evaluation of GSTM1 null polymorphism using Multiplex Polymerase Chain Reaction. RESULTS AND DISCUSSION: On comparing the prevalence of GSTM1 null polymorphism among the group with subjects with habits and no oral lesions, oral leukoplakia and oral SCC, it was observed that there was a statistically significant association between GSTM1 null polymorphism and the different groups (P < 0.01). CONCLUSION: The lack of GSTM1 activity would make the oral tissues more susceptible to action of tobacco carcinogens and to the development of a high-grade level of dysplasia in oral leukoplakia and thereby increases the susceptibility of lesion to undergo malignant changes.

Probabilistic multiple sclerosis lesion classification based on modeling regional intensity variability and local neighborhood information.

GOAL: In this paper, a fully automatic probabilistic method for multiple sclerosis (MS) lesion classification is presented, whereby the posterior probability density function over healthy tissues and two types of lesions (T1-hypointense and T2-hyperintense) is generated at every voxel. METHODS: During training, the system explicitly models the spatial variability of the intensity distributions throughout the brain by first segmenting it into distinct anatomical regions and then building regional likelihood distributions for each tissue class based on multimodal magnetic resonance image (MRI) intensities. Local class smoothness is ensured by incorporating neighboring voxel information in the prior probability through Markov random fields. The system is tested on two datasets from real multisite clinical trials consisting of multimodal MRIs from a total of 100 patients with MS. Lesion classification results based on the framework are compared with and without the regional information, as well as with other state-of-the-art methods against the labels from expert manual raters. The metrics for comparison include Dice overlap, sensitivity, and positive predictive rates for both voxel and lesion classifications. RESULTS: Statistically significant improvements in Dice values ( ), for voxel-based and lesion-based sensitivity values ( ), and positive predictive rates ( and respectively) are shown when the proposed method is compared to the method without regional information, and to a widely used method [1]. This holds particularly true in the posterior fossa, an area where classification is very challenging. SIGNIFICANCE: The proposed method allows us to provide clinicians with accurate tissue labels for T1-hypointense and T2-hyperintense lesions, two types of lesions that differ in appearance and clinical ramifications, and with a confidence level in the classification, which helps clinicians assess the classification results.

Immunogenicity of Escherichia coli expressed envelope 2 protein of Chikungunya virus.

Chikungunya fever, a re-emerging infection, is an arthropod-borne viral disease prevalent in different parts of the world, particularly Africa and South East Asia. Chikungunya virus envelope 2 protein is involved in binding to host receptors and it contains specific epitopes that elicit virus neutralizing antibodies. A highly immunogenic, recombinant Chikungunya virus envelope 2 protein was produced by bioreactor in Escherichia coli for development of a suitable diagnostic and vaccine candidate. This protein was refolded and further purified to achieve biologically active protein. The biological function of refolded and purified recombinant envelope 2 protein of Chikungunya virus was confirmed by its ability to generate envelope 2 specific antibodies with high titers in animal models. These findings suggest that recombinant envelope 2 protein of Chikungunya virus in combination with compatible adjuvant is highly immunogenic. Thus, recombinant envelope 2 protein can be a potential diagnostic reagent and vaccine candidate against Chikungunya virus infection.