Expression of short hairpin RNA (shRNA) targeting AC2 gene of Mungbean yellow mosaic India virus (MYMIV) reduces the viral titre in soybean.

Mungbean yellow mosaic India virus (MYMIV) belonging to the family Geminiviridae and the genus Begomovirus is a severe pathogen of tropical legumes including soybean. The absence of genetically mapped loci conferring resistance together with the genetic diversity of begomoviruses infecting soybean warrants the utilization of RNA interference (RNAi) technology to develop virus resistance. However, viral suppressors of RNAi (VSRs) reduce the effectiveness of RNA silencing. Here, we report the effectiveness of Agrobacterium-mediated transient expression of shRNA, targeting a conserved region of AC2 ORF (a VSR) of MYMIV, in conferring virus resistance in soybean. Transient expression of shRNA showed progressive reduction of the viral titre estimated by the MYMIV-derived AC2 gene copy numbers from the initial inoculum by approximately 80-fold 20 days post-application. In addition, the newly emerging leaves exhibited symptom recovery. Thus, this study proves that AC2 of MYMIV is a potent target gene for obtaining RNAi-mediated virus resistance in soybean. Agro-infiltration-based delivery of shRNA was an efficient means of gene silencing and could pave way for the development of transgenic virus-resistant soybean genotype.

A Quest for Accuracy: Evaluation of The Paris System in Diagnosis of Urothelial Carcinomas.

INTRODUCTION: Urine cytology is an important screening tool of patients for urothelial carcinoma (UC) and follow-up of patients with treated disease. Ease of procurement, cost-effectiveness, and lower turnaround time are the major advantages. OBJECTIVE: To compare current system of reporting (CSR) at our institute with The Paris System (TPS) and analyze utility of urine cytology based on TPS reporting in correlation with urine culture and histopathology. MATERIALS AND METHODS: One-year retrospective study of 90 cases was undertaken wherein cases presenting with painless hematuria and clinically suspicious of UC were included. Urine cytology slides were reviewed and reported with TPS guidelines. These findings were correlated with histopathological diagnosis and urine culture as indicated. Statistical analysis was done using SPSS 17 software. RESULTS: With TPS guidelines, 11.1% and 5.6% cases were reported as high-grade UC (HGUC) and low-grade urothelial neoplasm (LGUN), respectively. Suspicious for HGUC category included 17.8% of cases. The rate of reporting "atypical urothelial cells (AUC)" was significantly lower (11.1%) with TPS on comparison with CSR (16.7%). Histopathological correlation of positive predictive value for HGUC was better (100%) on using TPS when compared with CSR (64.3%). Among 11 cases with microbial growth on urine culture, 9.1% were reported as atypical. Sensitivity and accuracy of TPS in detecting UC were 83.3% and 86.52%, respectively. Both were higher when compared with CSR. CONCLUSION: In comparison to CSR, criteria of TPS limit the AUC category and enhance the sensitivity and accuracy of detecting HGUC. Adopting TPS for urinary cytology will ensure uniformity and accuracy of HGUC diagnosis.

Quantification of a legume begomovirus to evaluate soybean genotypes for resistance to yellow mosaic disease.

Mungbean yellow mosaic India virus (MYMIV) infecting soybean and other legumes causes yellow mosaic disease (YMD). Evaluation of soybean genotypes for YMD resistance involves field screening at disease hot spots or in a protected environment using infectious clones or viruliferous whiteflies as sources of virus inocula. Development of efficient virus inoculation and quantification protocols to screen soybean genetic stocks against YMD is imperative for breeding resistant varieties. Binary plasmids harbouring complete, tandem dimeric genomic components DNA A and DNA B of MYMIV-soybean isolate were engineered. The infectivity of the clones was demonstrated in soybean genotypes JS335 and UPSM534 that display contrasting YMD resistance. As a follow-up, soybean germplasm lines, breeding lines, and representative cultivars that were initially screened at an YMD hot-spot were then subjected to Agrobacterium-based infection with MYMIV. Quantitative real time polymerase chain reaction (qRT-PCR) based copy number analysis of MYMIV genomic components allowed soybean genotypes to be classified into three discrete categories; resistant, moderately resistant and susceptible to the viral infection. Thus, a soybean germplasm disease screening system based on agro-infection and qRT-PCR based quantification of MYMIV was developed to facilitate breeding YMD resistant soybean. The implications of this study for obtaining YMD resistant soybean cultivars are discussed.

Invasive meningococcal disease without meningitis: a forgotten diagnosis.

Neisseria meningitidis, a Gram-negative diplococcus, is an uncommon cause of pneumonia. There have been only about 344 cases reported worldwide from 1906 to 2015. To our knowledge, there have been only 3 cases reported in the USA in the past 2 decades. We present a case of a 72-year-old male with a past medical history of severe COPD, obstructive sleep apnea, and stage I lung cancer status post-stereotactic body radiation therapy 1 year ago, who was admitted with a 6-day history of productive cough with yellowish sputum, shortness of breath, extreme myalgias, and fatigue. Chest X-ray revealed an infiltrative process in the left lower lung field and left-sided pleural effusion. Blood cultures grew beta-lactamase-negative N. meningitidis after 24 hours. Our patient was initially treated with broad-spectrum antibiotics, which were later switched to amoxicillin to complete a total of 14 days of antibiotics. Diagnosing meningococcal pneumonia requires a high level of suspicion, as sputum cultures may be falsely positive due to asymptomatic carriage of the organism in the upper respiratory tract in up to 10% of outpatient population. We highlight this case as early recognition and treatment is critical. The case fatality rate for N. meningitidis pneumonia has been reported to be higher compared with meningococcal meningitis.

Fipronil-induced genotoxicity and DNA damage in vivo: Protective effect of vitamin E.

Fipronil, an insecticide of the phenylpyrazole class has been classified as a carcinogen by United States Environmental Protection Agency, yet very limited information is available about its genotoxic effects. Adult male and female animals were gavaged with various doses of fipronil (2.5, 12.5, and 25 mg/kg body weight (bw)) to evaluate micronucleus test (mice), chromosome aberration (CA), and comet assay (rats), respectively. Cyclophosphamide (40 mg/kg bw; intraperitoneal) was used as positive control. Another group of animals were pretreated with vitamin E orally (400 mg/kg bw) for 5 days prior to administration of fipronil (12.5 mg/kg). Fipronil exposure in both male and female mice caused significant increase in the frequency of micronuclei (MN) in polychromatic erythrocytes. Similarly, structural CAs in bone marrow cells and DNA damage in the lymphocytes was found to be significantly higher in the male and female rats exposed to fipronil as compared to their respective controls. The average degree of protection (male and female animals combined together) shown by pretreatment of vitamin E against fipronil-induced genotoxicity was 63.28%: CAs; 47.91%: MN formation; and 74.70%: DNA damage. Findings of this study demonstrate genotoxic nature of fipronil regardless of gender effect and documents protective role of vitamin E.

Erasure of fear memories is prevented by Nogo Receptor 1 in adulthood.

Critical periods are temporary windows of heightened neural plasticity early in development. For example, fear memories in juvenile rodents are subject to erasure following extinction training, while after closure of this critical period, extinction training only temporarily and weakly suppresses fear memories. Persistence of fear memories is important for survival, but the inability to effectively adapt to the trauma is a characteristic of post-traumatic stress disorder (PTSD). We examined whether Nogo Receptor 1 (NgR1) regulates the plasticity associated with fear extinction. The loss of NgR1 function in adulthood eliminates spontaneous fear recovery and fear renewal, with a restoration of fear reacquisition rate equal to that of naive mice; thus, mimicking the phenotype observed in juvenile rodents. Regional gene disruption demonstrates that NgR1 expression is required in both the basolateral amygdala (BLA) and infralimbic (IL) cortex to prevent fear erasure. NgR1 expression by parvalbumin expressing interneurons is essential for limiting extinction-dependent plasticity. NgR1 gene deletion enhances anatomical changes of inhibitory synapse markers after extinction training. Thus, NgR1 robustly inhibits elimination of fear expression in the adult brain and could serve as a therapeutic target for anxiety disorders, such as PTSD.

Persistence of Corynebacterium diphtheriae in Delhi & National Capital Region (NCR).

Despite the introduction of mass immunization, diphtheria continues to play a major role as a potentially lethal infectious disease in many countries. Delay in the specific therapy of diphtheria may result in death and, therefore, accurate diagnosis of diphtheria is imperative. This study was carried out at National Centre for Disease Control (NCDC), Delhi, India, on samples of suspected diphtheria cases referred from various government hospitals of Delhi and neighbouring areas during 2012-2014. Primary identification of Corynebacterium diphtheriae was done by standard culture, staining and biochemical tests followed by toxigenicity testing by Elek's test on samples positive for C. diphtheriae. The results showed persistence of toxigenic C. diphtheriae in our community indicating the possibility of inadequate immunization coverage.

A rare case of isolated hydatid cyst of breast.

INTRODUCTION: Hydatid cyst of the breast is very rare. It is challenging to differentiate it from other tumoral lesions of the breast. Only few reports of breast hydatid cyst are published and majority of the reported cases have been diagnosed postoperatively as it is not possible to reach definitive diagnosis with clinical examination and radiological investigations only. PRESENTATION OF CASE: A 31-year old woman presented with a painless lump in the right breast since one year duration. On clinical examination, a non-mobile, firm lump was detected in the right breast associated with nipple retraction, but there was no axillary lymphadenopathy. This case was diagnosed as hydatid cyst incidentally during surgery from its gross appearance which mimics that of a liver hydatid cyst, normally common in this endemic area. DISCUSSION: Hydatid disease is a parasitic infection caused by the larval form of Echinococcus granulosus and seen endemically among sheep-raising communities. The breast can be a primary site or part of a disseminated hydatidosis. It might mimic fibroadenoma, phyllodes tumors, chronic abscesses, or even carcinoma. Preoperative diagnosis can be made by fine needle aspiration cytology. It also can be diagnosed by radiological or serologic means but neither of them is definitive. Surgery is the treatment of choice. CONCLUSION: Hydatid cyst of the breast is very uncommon but it should be included in differential diagnosis of breast lumps for patients living in endemic areas.

An evaluation of multimodal spinal cord monitoring in scoliosis surgery: a single centre experience of 354 operations.

PURPOSE: To evaluate the effectiveness of multimodal intraoperative neuromonitoring in the early detection of impending spinal cord injury during surgery for spinal deformity. METHOD: A retrospective review of prospectively collected data in 354 consecutive spinal deformity operations from June 2003 to October 2013. Patients were sub-grouped according to demographics, diagnosis and operative features. Post-operative neurological deficit was defined as either spinal cord, nerve root or transient deficit. RESULTS: Combined monitoring with SSEPs and MEPs was possible in 315 cases. The overall incidence of significant alerts was 7.1 % and overall permanent neurological deficit was 1.6 %. When results were collated, the overall combined sensitivity of multimodal monitoring was 100 % with a specificity of 99.3 %. CONCLUSIONS: Multimodal monitoring allows early detection of impending neurological deficit that is superior to a single monitoring modality. To achieve optimal use of monitoring, continuous communication between surgical, anaesthetic and neurophysiology teams are required. As a result of our experience we have incorporated in our consent procedure the discussion of monitoring and the possibility of needing to abandon the procedure, and completing in a staged fashion at a later date. We believe multimodal monitoring is the current gold standard for complex spinal deformity surgery.

Novel Na(+) doped Alq3 hybrid materials for organic light-emitting diode (OLED) devices and flat panel displays.

Pure and Na(+) -doped Alq3 complexes were synthesized by a simple precipitation method at room temperature, maintaining a stoichiometric ratio. These complexes were characterized by X-ray diffraction, Fourier transform infrared (FTIR), UV/Vis absorption and photoluminescence (PL) spectra. The X-ray diffractogram exhibits well-resolved peaks, revealing the crystalline nature of the synthesized complexes, FTIR confirms the molecular structure and the completion of quinoline ring formation in the metal complex. UV/Vis absorption and PL spectra of sodium-doped Alq3 complexes exhibit high emission intensity in comparison with Alq3 phosphor, proving that when doped in Alq3 , Na(+) enhances PL emission intensity. The excitation spectra of the synthesized complexes lie in the range 242-457 nm when weak shoulders are also considered. Because the sharp excitation peak falls in the blue region of visible radiation, the complexes can be employed for blue chip excitation. The emission wavelength of all the synthesized complexes lies in the bluish green/green region ranging between 485 and 531 nm. The intensity of the emission wavelength was found to be elevated when Na(+) is doped into Alq3 . Because both the excitation and emission wavelengths fall in the visible region of electromagnetic radiation, these phosphors can also be employed to improve the power conversion efficiency of photovoltaic cells by using the solar spectral conversion principle. Thus, the synthesized phosphors can be used as bluish green/green light-emitting phosphors for organic light-emitting diodes, flat panel displays, solid-state lighting technology - a step towards the desire to reduce energy consumption and generate pollution free light.

Synthesis and characterization of pure and Li⁺ activated Alq3 complexes for green and blue organic light emitting diodes and display devices.

Pure and Li(+)-doped Alq3 complexes were synthesized by simple precipitation method at room temperature, maintaining the stoichiometric ratio. These complexes were characterized by X-ray diffraction, ultraviolet-visible absorption and Fourier transform infrared and photoluminescence (PL) spectra. X-ray diffraction analysis reveals the crystalline nature of the synthesized complexes, while Fourier transform infrared spectroscopy confirm the molecular structure, the completion of quinoline ring formation and presence of quinoline structure in the metal complex. Ultraviolet-visible and PL spectra revealed that Li(+) activated Alq3 complexes exhibit the highest intensity in comparison to pure Alq3 phosphor. Thus, Li(+) enhances PL emission intensity when doped into Alq3 phosphor. The excitation spectra lie in the range of 383-456 nm. All the synthesized complexes other than Liq give green emission, while Liq gives blue emission with enhanced intensity. Thus, he synthesized phosphors are the best suitable candidates for green- and blue-emitting organic light emitting diode, PL liquid-crystal display and solid-state lighting applications.

A Unusual Case of Complicated Zenker's Diverticulum.

We report the surgical management of a case of Zenker's diverticulum in a 64 year old man, complicated by metallic nail penetration and surgical scarring due to previous surgery for cervical vertebral trauma.

Study of photophysical properties of different metal complexes of Alq3.

Different metal complexes of Alq3 (K(+) , Sr(2+) , Ca(2+) and Mg(2+) ) were synthesized by the precipitation method. The characterization of photoluminescence showed that presence of Mg(2+) ion enhances photoluminescence of Alq3 phosphor. The emission spectra are observed at 560 nm when excited at a wavelength of 440 nm. The phosphor is excited at a longer wavelength in the blue region of small energy, so that it could be used as lamp phosphor. It is observed that the prepared phosphor AlMgq5 is more suitable than Alq3. The ionic radii of K(+) , Sr(2+) , Ca(2+) and Mg(2+) ions are in decreasing order. Therefore, the remarkable properties of AlMgq5 could be considered as promising materials as opto- or optoelectronic materials.

Chronic, noninvasive glucocorticoid administration suppresses limbic endocannabinoid signaling in mice.

Limbic endocannabinoid signaling is known to be sensitive to chronic stress; however, studies investigating the impact of prolonged exposure to glucocorticoid hormones have been limited by the concurrent exposure to the stress of daily injections. The present study was designed to examine the effects of a noninvasive approach to alter plasma corticosterone (CORT) on the endocannabinoid system. More precisely, we explored the effects of a 4-week exposure to CORT dissolved in the drinking water of mice (100 µg/ml) and measured cannabinoid CB(1) receptor binding, endocannabinoid content, activity of the endocannabinoid degrading enzyme fatty acid amide hydrolase (FAAH), and mRNA expression of both the CB(1) receptor and FAAH in both the hippocampus and amygdala. Our data demonstrate that CORT decreases CB(1) receptor binding site density in both the hippocampus and amygdala and also reduced anandamide (AEA) content and increased FAAH activity within both structures. These changes in both CB(1) receptor binding and FAAH activity were not accompanied by changes in mRNA expression of either the CB(1) receptor or FAAH in either brain region. Interestingly, our CORT delivery regimen significantly increased 2-AG concentrations within the hippocampus, but not the amygdala. Collectively, these data demonstrate that the confounder of injection stress is sufficient to conceal the ability of protracted exposure to glucocorticoids to reduce CB(1) receptor density and augment AEA metabolism within limbic structures.

Isolation and Characterization of Trichoderma spp. for Antagonistic Activity Against Root Rot and Foliar Pathogens.

Trichoderma, soil-borne filamentous fungi, are capable of parasitising several plant pathogenic fungi. Twelve isolates of Trichoderma spp. isolated from different locations of South Andaman were characterized for their cultural, morphological and antagonistic activity against soil borne and foliar borne pathogens. The sequencing of these isolates showed seven different species. The isolates revealed differential reaction patterns against the test pathogens viz., Sclerotium rolfsii, Colletotrichum gloeosporioides and C. capsici. However, the isolates, TND1, TWN1, TWC1, TGD1 and TSD1 were most effective in percentage inhibition of mycelial growth of test pathogens. Significant chitinase and ß-1,3-glucanase activities of all Trichoderma isolates has been recorded in growth medium. T. viride was found with highest chitinase whereas T. harzianum was recorded with highest ß-1,3-glucanase activities.

Ossicular chain status in chronic suppurative otitis media in adults.

This study was conducted to find out the status of the ossicles in cases of chronic suppurative otitis media (CSOM). One hundred and fifty cases of CSOM, who underwent surgery, were included and their intra-operative ossicular chain findings noted. Ossicular erosion was found to be much more common in unsafe CSOM than in safe CSOM. Malleus was found to be the most resistant ossicle to erosion whereas incus was found to be the most susceptible.

Combined Cervical and Video-assisted Thoracoscopic Thyroidectomy (CAVATT): A Simplified and Innovative Approach for Goiter with Posterior Mediastinal Extension.

We report a case of benign multinodular goiter with right sided posterior mediastinal extension managed by a simplified anaesthetic and surgical protocol. Single lumen endotracheal tube was used for intubation. Access to mediastinum was obtained by 2 working ports in 2nd and 4th intercostal spaces. Mediastinal extension is dissected thoracoscopically and delivered cervically. Post operative course was uneventful with no pulmonary and surgical morbidity.