A detailed model and Monte Carlo simulation for predicting DIP genome length distribution in baculovirus infection of insect cells.

Baculoviruses have enormous potential for use as biopesticides to control insect pest populations without the adverse environmental effects posed by the widespread use of chemical pesticides. However, continuous baculovirus production is susceptible to DNA mutation and the subsequent production of defective interfering particles (DIPs). The amount of DIPs produced and their genome length distribution are of great interest not only for baculoviruses but for many other DNA and RNA viruses. In this study, we elucidate this aspect of virus replication using baculovirus as an example system and both experimental and modeling studies. The existing mathematical models for the virus replication process consider DIPs as a lumped quantity and do not consider the genome length distribution of the DIPs. In this study, a detailed population balance model for the cell-virus culture is presented, which predicts the genome length distribution of the DIP population along with their relative proportion. The model is simulated using the kinetic Monte Carlo algorithm, and the results agree well with the experimental results. Using this model, a practical strategy to maintain the DIP fraction to near to its maximum and minimum limits has been demonstrated.

A structured review of baculovirus infection process: integration of mathematical models and biomolecular information on cell-virus interaction.

The baculovirus expression vector system (BEVS) is an emerging tool for the production of recombinant proteins, vaccines and bio-pesticides. However, a system-level understanding of the complex infection process is important in realizing large-scale production at a lower cost. The entire baculovirus infection process is summarized as a combination of various modules and the existing mathematical models are discussed in light of these modules. This covers a systematic review of the present understanding of virus internalization, viral DNA replication, protein expression, budded virus (BV) and occlusion-derived virus (ODV) formation, few polyhedral (FP) and defective interfering particle (DIP) mutant formation, cell cycle modification and apoptosis during the viral infection process. The corresponding theoretical models are also included. Current knowledge regarding the molecular biology of the baculovirus/insect cell system is integrated with population balance and mass action kinetics models. Furthermore, the key steps for simulating cell and virus densities and their underlying features are discussed. This review may facilitate the further development and refinement of mathematical models, thereby providing the basis for enhanced control and optimization of bioreactor operation.

Human papilloma virus (HPV) infection leads to the development of head and neck lesions but offers better prognosis in malignant Indian patients.

Head and neck cancers constitute a multifactorial global disease burden and are associated with human papilloma virus (HPV) as a possible risk factor. The aim of the study is to understand the relationship between HPV and the development of head and neck lesions in Indian patients. To this end, frequency of HPV was assessed in relation to different demographic and etiological features and correlated with patient survival. The prevalence of HPV significantly increased from mild dysplastic lesions (43.6%) to head and neck squamous cell carcinoma (HNSCC) stage IV (68.5%) with HPV 16 being pre-dominant in both dysplasia (43.8%) and HNSCC (61.5%). Similar trend was observed in increasing grades of the tumour. In invasive lesions, patients aged below the median age of onset showed significantly higher occurrence of HPV than those above it. Patients harbouring HPV showed a significantly better survival irrespective of age of onset. Likewise, better survival was observed in tobacco habit negative/HPV-positive patients, and as reflected in both univariate and multivariate analysis. Majority of the HPV 16-positive samples showed moderate/high nuclear expression of HPV E6 and E7 proteins in tumours and respective basal layer of adjacent normal tissues. Thus, our data indicate that frequent HPV infection, along with tobacco habit, is a pre-requisite factor for the development of HNSCC of Indian patients but offers a better survival even during tobacco usage, implicating its diagnostic and prognostic importance.

WITHDRAWN: Differential molecular signature alterations of RBSP3, LIMD1 and CDC25A in normal oral epithelium during oral tumorigenesis.

The Accepted Manuscript version of this article (published on 1 June 2016) was withdrawn on 24 November 2017 at the request of the authors.

MicroRNA profiling of cisplatin-resistant oral squamous cell carcinoma cell lines enriched with cancer-stem-cell-like and epithelial-mesenchymal transition-type features.

Oral cancer is of major public health problem in India. Current investigation was aimed to identify the specific deregulated miRNAs which are responsible for development of resistance phenotype through regulating their resistance related target gene expression in oral squamous cell carcinoma (OSCC). Cisplatin-resistant OSCC cell lines were developed from their parental human OSCC cell lines and subsequently characterised. The resistant cells exhibited enhanced proliferative, clonogenic capacity with significant up-regulation of P-glycoprotein (ABCB1), c-Myc, survivin, ß-catenin and a putative cancer-stem-like signature with increased expression of CD44, whereas the loss of E-cadherin signifies induced EMT phenotype. A comparative analysis of miRNA expression profiling in parental and cisplatin-resistant OSCC cell lines for a selected sets (deregulated miRNAs in head and neck cancer) revealed resistance specific signature. Moreover, we observed similar expression pattern for these resistance specific signature miRNAs in neoadjuvant chemotherapy treated and recurrent tumours compared to those with newly diagnosed primary tumours in patients with OSCC. All these results revealed that these miRNAs play an important role in the development of cisplatin-resistance mainly through modulating cancer stem-cell-like and EMT-type properties in OSCC.

Frequent inactivation of SLIT2 and ROBO1 signaling in head and neck lesions: clinical and prognostic implications.

OBJECTIVE: The protein SLIT2 and its receptor ROBO1 regulate different cellular processes, such as proliferation, apoptosis, and migration. In this study our aim is to understand the alterations of these genes during development of head and neck squamous cell carcinoma (HNSCC). MATERIALS AND METHODS: First, molecular alterations of the genes were analyzed in 30 dysplastic lesions, 128 primary HNSCC samples, and 1 HNSCC cell line. Then alterations were correlated with mRNA expression (n = 22) and protein expression (n = 29). Finally, the alterations were correlated with different clinicopathologic parameters and clinical outcomes of the patients. RESULTS: ROBO1 had a comparatively high frequency of deletion (28.5%-54.2%) from dysplastic lesions and subsequent clinical stages than did SLIT2 (16.6-27%). On the contrary, SLIT2 had a high frequency (56.6%-81.2%) of promoter methylation from dysplastic lesions onward compared with ROBO1 (20%-32.8%). Interestingly, alterations of SLIT2 and ROBO1 were high in dysplastic lesions (80%), followed by comparable frequencies (92.5%-95.3%) in subsequent stages of tumor. Alterations of these genes showed concordance with their mRNA/protein expression and significant association with poor patient outcome. CONCLUSIONS: Our data suggest that inactivation of SLIT2 and/or ROBO1 is one of the early events in development of dysplastic lesions of head and neck and has prognostic importance.

Synthesis and in situ observation of 3D superlattices of gold nanoparticles using oil-in-water emulsion.

In this work oil-in-water emulsion has been successfully used as a confined environment to grow 3D superlattices of gold nanoparticles. The superlattices were grown from 5 nm uniform gold nanoparticles using slow destabilization method. The confined environment was created by forming a stable emulsion where the gold colloid suspended in toluene was used as oil phase. Superlattices were also formed in bulk solution using the same slow destabilization method. A comparative study reveals that compact superlattices form more readily inside the emulsion drops as compared to bulk precipitation. The unstable colloid (in bulk or as emulsion) was aged for various periods at 5 °C to form more compact superlattices. The best superlattices with sharp corners are observed when the superlattices are formed inside the emulsion and aged for a month. Two key parameters, the incubation temperature and anti-solvent concentration, are optimized to obtain larger superlattices with sharp features. A new method is also demonstrated for in situ observation of superlattice formation using an optical microscope.

Anaesthetic management of a child with large occipital meningomyelocele.

A meningomyelocele is herniation of meninges along with neural elements through a congenital defect in the cranium or vertebral column. Reported incidence of occipital meningomyelocele is 1 in 5000 live births. Children with meningomyelocele may have varying degrees of sensory and motor deficits and other important associated congenital defects with clinical features of brain stem compression. The major anaesthetic challenges associated with myelomeningocoele repair includes difficulty in securing airway, intra-operative prone positioning, accurate assessment of blood loss and prevention of hypothermia.

The effect of dexmedetomidine infusion on propofol requirement for maintenance of optimum depth of anaesthesia during elective spine surgery.

BACKGROUND: Maintenance of adequate depth of anaesthesia in spine surgery is vital to prevent awareness, to reduce stress response and possible autonomic instability frequently associated with spine surgery. Dexmedetomidine, a α2-adrenoceptor agonist with analgesic and sedative adjuvant property has been found to reduce dose requirement of multiple anaesthetic agents both for induction and during the maintenance of anaesthesia. AIM: The aim of this study is to observe the effect of dexmedetomidine, on the requirement of propofol for induction and maintenance of adequate depth of anaesthesia during spine surgery. METHODS: It was a prospective, randomised, double-blinded, parallel group, placebo controlled and open-lebel study in tertiary care hospital. A total of 70 patients aged 20-60 years, American Society of Anaesthesiologists GradeI and II, scheduled for elective spine surgery were randomly allocated into two groups. Each patient of Group D (n=35) received an initial loading dose of dexmedetomidine at 1 µg/kg over 10 min, started 15 min before induction of anaesthesia followed by an infusion at a rate of 0.2 µg/kg/h. Patients of Group P (n=35) received the same volume of 0.9% normal saline solution as placebo. Requirement of propofol at induction and during maintenance was calculated maintaining bispectral index between 40 and 60. P<0.05 was considered to be statistically significant. RESULTS: Mean requirement of propofol was found to be lessened by 48.08% and 61.87% for induction and maintenance of anaesthesia respectively while using dexmedetomidine. CONCLUSION: Administration of dexmedetomidine significantly reduces the requirement of propofol while maintaining desired depth of anaesthesia without any significant complication.

Inactivation of 9q22.3 tumor suppressor genes predict outcome for patients with head and neck squamous cell carcinoma.

AIM: This study examined the prognostic significance of candidate tumor suppressor genes (TSGs) PHD finger protein-2 (PHF2), Fanconi anaemia complementation group C (FANCC) and human homologue of Drosophila patched gene (PTCH1), in head and neck squamous cell carcinoma (HNSCC) treated primarily with surgery, or surgery followed by adjuvant radiotherapy. PATIENTS AND METHODS: Eighty-four patients with HNSCC were followed-up for recurrence/death for up to five years after diagnosis. Molecular alterations (deletion/methylation) of TSGs and human papilloma virus (HPV) status were determined in previous studies of our group. Statistical analyses of correlation of genetic alterations with treatment response and survival were carried out. RESULTS: Alterations of FANCC and PTCH1 were significantly associated with locoregional recurrence/death. In the surgery with adjuvant radiotherapy-group (n=56), patients showing alterations in FANCC and in PTCH1 were seven- and six-times, respectively, more likely to have locoregional recurrence compared to those with no alterations. In addition, the presence of alterations of both FANCC and PTCH1 had remarkable prognostic significance. CONCLUSION: FANCC and PTCH1 alterations might be used as molecular markers for prognosis and to develop strategies for effective treatment planning.

Association of FANCC and PTCH1 with the development of early dysplastic lesions of the head and neck.

BACKGROUND: Alteration of chromosome 9q22.3 region is an early and frequent event in head and neck squamous cell carcinoma (HNSCC). The aim of this study was to understand the association of candidate tumor suppressor genes PHF2, FANCC, PTCH1, and XPA located in this region in the development of HNSCC. METHODS: The alterations (deletion, promoter methylation, mutation, expression) of these genes were analyzed in 65 dysplastic head and neck lesions and 84 primary HNSCC samples. Clinicopathologic correlations were made with alterations of the genes. RESULTS: Overall alterations (deletion, promoter methylation) of FANCC and PTCH1 were high in mild dysplasia and comparable in subsequent stages of tumor progression. However, PHF2 alteration was low in mild dysplasia, but increased in moderate and severe dysplasias. Alterations (deletion, promoter methylation) of FANCC and PTCH1 showed association with each other. Two novel mutations in GLI binding sites of PTCH1 promoter and a novel microsatellite marker hmPTCH1 with four alleles at immediate upstream of the gene were identified. In a case-control study, the (CGG)7 allele of hmPTCH1 was found to be susceptible for HNSCC development. Concordance was seen in the expression (RNA, protein) of these genes with their molecular alterations. CONCLUSIONS: Alterations of FANCC and PTCH1 could be used as molecular marker for early diagnosis and prognosis of HNSCC.

Anaesthetic management of a patient with motor neuron disease posted for microlaryngeal surgery.

Motor neuron disease (MND) typically affects upper and lower motor neurons without any sensory involvement. Regional anaesthesia is preferred in motor neuron disease patients to avoid aspiration and possibility of prolonged postoperative ventilation following general anaesthesia. As microlaryngeal surgery in a motor neuron disease patient needed airway protection, it was decided to perform the surgery under general anaesthesia with awake intubation following airway block and avoiding muscle relaxant agents completely to minimise unnecessary complications. The patient was allowed spontaneous breathing while maintaining anaesthesia and stable haemodynamic condition with combination of anaesthetic and analgesic agents accordingly throughout the operation.

CHARGE Association.

We present here a case of 17-year-old boy from Kolkata presenting with obesity, bilateral gynecomastia, mental retardation, and hypogonadotrophic hypogonadism. The patient weighed 70 kg and was of 153 cm height. Facial asymmetry (unilateral facial palsy), gynecomastia, decreased pubic and axillary hair, small penis, decreased right testicular volume, non-palpable left testis, and right-sided congenital inguinal hernia was present. The patient also had disc coloboma, convergent squint, microcornea, microphthalmia, pseudohypertelorism, low set ears, short neck, and choanalatresia. He had h/o VSD repaired with patch. Laboratory examination revealed haemoglobin 9.9 mg/dl, urea 24 mg/dl, creatinine 0.68 mg/dl. IGF1 77.80 ng/ml (decreased for age), GH <0.05 ng/ml, testosterone 0.25 ng/ml, FSH-0.95 µIU/ml, LH 0.60 ΅IU/ml. ACTH, 8:00 A.M cortisol, FT3, FT4, TSH, estradiol, DHEA-S, lipid profile, and LFT was within normal limits. Prolactin was elevated at 38.50 ng/ml. The patient's karyotype was 46XY. Echocardiography revealed ventricularseptal defect closed with patch, grade 1 aortic regurgitation, and ejection fraction 67%. Ultrasound testis showed small right testis within scrotal sac and undescended left testis within left inguinal canal. CT scan paranasal sinuses revealed choanalatresia and deviation of nasal septum to the right. Sonomammography revealed bilateral proliferation of fibroglandular elements predominantly in subareoalar region of breasts. MRI of brain and pituitary region revealed markedly atrophic pituitary gland parenchyma with preserved infundibulum and hypothalamus and widened suprasellar cistern. The CHARGE association is an increasingly recognized non-random pattern of congenital anomalies comprising of coloboma, heart defect, choanal atresia, retarded growth and development, genital hypoplasia, ear abnormalities, and/or deafness.[1] These anomalies have a higher probability of occurring together. In this report, we have described a boy with CHARGE association.

A rare occurrence of isolated neutropenia with valproic acid: a case report.

Valproic acid, a conventional antiepileptic drug, is also used in the treatment of bipolar disorder, to provide rapid stabilization of the patient. Although thrombocytopenia is a potential threat, occurrence of suppression of the other cell lines in the bone marrow is also found rarely. Here in this study we report a case of a 28 year old female treated with valproic acid and risperidone for bipolar disorder, who developed isolated neutropenia within 6 months of commencement of therapy that got reversed soon after termination of valproic acid therapy. This finding lays further emphasis on regular monitoring of all cell lines in blood for better management and to prevent any serious bone marrow toxicity in the patients.

A comparative study of duration of postoperative analgesia between epidural bupivacaine and epidural clonidine plus bupivacaine in lumbar laminectomy surgery under general anaesthesia.

A randomised prospective double-blind placebo controlled study was undertaken in 60 patients of ASA-I and II, scheduled for lumbar laminectomy under general anaesthesia using thiopentone, fentanyl, artracurium, N2O/O2 and isoflurane. After obtaining written Informed consent from all patients, they were randomly allocated to two equal groups ie, group A (n = 30) and group B (n = 30). Group A patients received clonidine 0.5 ml (75 microg) plus 5 ml 0.25% bupivacaine and group B patients received 5 ml 0.25% bupivacaine plus 0.5 ml of normal saline (NS) as a control through epidural route placed by the surgeon at the closure of the surgery. All the vital parameters were recorded at the time of epidural catheter placement and administering drugs and at 10 minutes interval thereafter till the patients were reversed with neostigmine and glycopyrrolate and then in the postanaesthetic care unit (PACU) at 15 minutes interval till the end of the study. All patients received injection diclofenac 75 mg IM as soon as they complained of pain or when the VAS score became > or = 4cm. The study ended when patients received injection diclofenac 75 mg IM as rescue analgesic. Any feature of motor block, sedation score and event of urinary retention were recorded in the PACU. Duration of postoperative analgesia was taken as the primary outcome and measured from the time of regaining full consciousness following surgery till receiving first injection of rescue analgesic in the PACU. Duration of postoperative analgesia was (546.36 +/- 11.55 minutes) in group A compared to (240.15 +/- 07.32 minutes) in group B. No clinically significant difference was found in heart rate, blood pressure, respiratory rate, oxygen saturation and motor blockade. Sedation score was slightly higher in group A, which was not significant. So small dose of clonidine (75 microg) as an adjuvant to 5 ml 0.25% bupivacaine in epidural route following lumbar laminectomy significantly prolonged postoperative analgesia and improved patient satisfaction without any clinically significant adverse reaction.

A randomized controlled trial of oral zinc in acute pneumonia in children aged between 2 months to 5 years.

OBJECTIVE: To evaluate the effectiveness and safety of zinc supplementation as adjuvant in treatment of pneumonia. METHODS: Ninety-eight children with acute bacterial pneumonia, aged between 2 months to 5 years, were studied in a randomized controlled single blind design. They received either zinc supplementation, as zinc acetate syrup, or placebo, as vitamin B-complex syrup, for 14 days, concomitantly with antimicrobial treatment (49 per group). Chest radiograph and blood tests were done for confirmation of diagnosis and severity of pneumonia was assessed by breathing rate, chest in-drawing and body temperature. Potentially immunosuppressed children or those with serious comorbidity were excluded. Follow-up was done daily while subjects were admitted (generally 7 days) and the final assessment made on the 14th day on out-patient basis. RESULTS: Children enrolled in zinc and placebo groups were of comparable age [17 ± 10 and 10 ± 30 months (median ± interquartile range) respectively] and sex distribution [34 (69.4%) vs 31 (63.3%) males respectively]. Duration of illness at diagnosis was also comparable. Patients supplemented with zinc showed no difference in clinical cure rate at 14 days when compared with placebo. Fast breathing was present after 1 wk of treatment in 49% subjects in zinc supplemented vs 43% on placebo (p = 0.685). There was also no difference in breathing rate at study end. Regarding fever, the mean temperature was <99°F in both groups at study end. Hemoglobin, total leukocyte count, standard liver function tests and creatinine showed no difference between groups either at baseline or at study end. There were no treatment emergent adverse events attributable to zinc. CONCLUSIONS: Though well tolerated; the addition of zinc does not improve symptom duration or cure rate in acute bacterial pneumonia in under-five children.

Peri-operative flexometallic endotracheal tube malfunction--an unusual complication.

Increased difficulty was encountered during manual ventilation following change of position from supine to prone in a patient intubated with a fresh flexometallic cuffed endotracheal tube, posted for elective lumbar discectomy. Prompt Identification of the problem was made and managed with replacement of the damaged tube with a new endotracheal tube in supine position. Following proper securing of the airway and repositioning of the patient, rest of the surgery and anaesthetic maintenance were uneventful. The cause of this unusual complication was ultimately found to be an obstruction along the length of lumen of the culprit endotracheal tube which was maximum at its proximal end by herniation of the deformed channel of the cuff with filled air generated due to backpressure from the tracheal cuff that might have been produced due to manufacturing defect.

Extreme levels of underweight and stunting among pre-adolescent children of low socioeconomic class from Madhyamgram and Barasat, West Bengal, India.

A cross-sectional study on 1206 children (788 boys, 618 girls) aged 1-12 years, belonging to low socioeconomic status, of Barasat and Madhyamgram, West Bengal, India, was undertaken to investigate age and sex variations in height and weight. It also evaluated the levels of underweight and stunting among them. Anthropometric measurements included weight and height. Weight-for-age (WA) and height-for-age (HA) <-2 z-scores were used to evaluate underweight (UW) and stunting (ST), respectively, following the National Center for Health Statistics (NCHS) Guidelines. Results showed that boys aged 4 and 11 years were significantly heavier than girls of corresponding ages. Boys aged 7 years were significantly taller than girls of the same age. Significant (p < 0.001) age differences existed in mean weight and height in boys (weight: F = 336.762; height: F = 565.160) as well as girls (weight: F = 275.320; height = 498.715). Results also revealed that the mean z-scores of WA (WAZ) and HA (HAZ) were less than (negative values) those of NCHS for both sexes at all ages. The overall (age combined) rate of UW was 60.4% and 51.3%; while that of ST was 51.7% and 48.4%, in boys and girls, respectively. Based on World Health Organization classification of severity of malnutrition, the prevalence of UW and ST were very high (> or = 30%) in both sexes. In conclusion, this study demonstrated that the prevalence of UW and ST were very high among the subjects. Since the nutritional status of the subjects is not satisfactory, there is need for immediate supplementary nutrition.