Characterisation of novel microbial strains Proteus mirabilis and Bordetella avium for heavy metal bioremediation and dye degradation.

The entitled study focuses on exploring the microbial diversity and its applicability in the remediation of metal contaminated soil using microbes, which is a reliable and cost effective technique. Tungsten enriched soil of Kuhi-Agargaon-Khobna region (Nagpur, India) were analysed by XRF method to detect heavy metals. The traditional microbiological techniques were used to isolate tungsten tolerant microbes. Applicability of these microbes in bioremediation and Azo dye degradation was mainly studied. The two novel bacterial strains, Proteus mirabilis (RS2K) and Bordetella avium (RS3K), were isolated and identified to show the tolerance to tungsten, using 16S rDNA and phylogenetic analysis. These novel strains have also shown the tolerance to other metallic salts viz., (sodium) tungsten, tungstic acid, ammonium metaparatungstate, mercuric chloride, cobalt chloride and azo dye. These microbes were found to accumulate tungsten intracellularly as confirmed through ICP-MS and SEM-EDS analyses. Microbes exhibited well-equipped cellular mechanisms for metal tolerance to survive in heavy metal-laden ecology. Current study contains substantial potential in bioleaching of heavy metals and green mining along with Nano bioremediation for heavy metal pollution.

Recent advances on mycotic keratitis caused by dematiaceous hyphomycetes.

Dematiaceous hyphomycetes (DH) are darkly pigmented fungi ubiquitously found all over the world as plant pathogens and saprophytes, and many of the members of this group have emerged as opportunistic pathogens. These fungi are responsible for a wide variety of infections including mycotic keratitis, which is considered as one of the major causes of corneal blindness, particularly in tropical and subtropical countries with an annual global burden of about 1 000 000 patients. The infection is more common in workers working in an outdoor environment. Moreover, trauma is found to be the most important predisposing cause of mycotic keratitis. Considerable delay in diagnosis and scarcity of effective pharmacological drugs are the major factors responsible for increased morbidity and visual impairment. Considering the crucial role of DH in mycotic keratitis, in the present review, we have focused on major DH with special emphasis on their pathogenicity, diagnosis and treatment strategies.

Heteroplasmy due to coexistence of mtCOI haplotypes from different lineages of the Thrips tabaci cryptic species group.

Heteroplasmy is the existence of multiple mitochondrial DNA haplotypes within the cell. Although the number of reports of heteroplasmy is increasing for arthropods, the occurrence, number of variants, and origins are not well studied. In this research, the occurrence of heteroplasmy was investigated in Thrips tabaci, a putative species complex whose lineages can be distinguished by their mitochondrial DNA haplotypes. The results from this study showed that heteroplasmy was due to the occurrence of mitochondrial cytochrome oxydase I (mtCOI) haplotypes from two different T. tabaci lineages. An assay using flow cytometry and quantitative real-time PCR was then used to quantify the per cell copy number of the two mtCOI haplotypes present in individuals exhibiting heteroplasmy from nine geographically distant populations in India. All of the T. tabaci individuals in this study were found to exhibit heteroplasmy, and in every individual the per cell copy number of mtCOI from lineage 3 comprised 75-98% of the haplotypes detected and was variable among individuals tested. There was no evidence to suggest that the presense of lineage-specific haplotypes was due to nuclear introgression; however, further studies are needed to investigate nuclear introgression and paternal leakage during rare interbreeding between individuals from lineages 2 and 3.

Drug-sensitive FGFR3 mutations in lung adenocarcinoma.

Background: Lung cancer is the leading cause of cancer-related deaths across the world. In this study, we present therapeutically relevant genetic alterations in lung adenocarcinoma of Indian origin. Materials and methods: Forty-five primary lung adenocarcinoma tumors were sequenced for 676 amplicons using RainDance cancer panel at an average coverage of 1500 × (reads per million mapped reads). To validate the findings, 49 mutations across 23 genes were genotyped in an additional set of 363 primary lung adenocarcinoma tumors using mass spectrometry. NIH/3T3 cells over expressing mutant and wild-type FGFR3 constructs were characterized for anchorage independent growth, constitutive activation, tumor formation and sensitivity to FGFR inhibitors using in vitro and xenograft mouse models. Results: We present the first spectrum of actionable alterations in lung adenocarcinoma tumors of Indian origin, and shows that mutations of FGFR3 are present in 20 of 363 (5.5%) patients. These FGFR3 mutations are constitutively active and oncogenic when ectopically expressed in NIH/3T3 cells and using a xenograft model in NOD/SCID mice. Inhibition of FGFR3 kinase activity inhibits transformation of NIH/3T3 overexpressing FGFR3 constructs and growth of tumors driven by FGFR3 in the xenograft models. The reduction in tumor size in the mouse is paralleled by a reduction in the amounts of phospho-ERK, validating the in vitro findings. Interestingly, the FGFR3 mutations are significantly higher in a proportion of younger patients and show a trend toward better overall survival, compared with patients lacking actionable alterations or those harboring KRAS mutations. Conclusion: We present the first actionable mutation spectrum in Indian lung cancer genome. These findings implicate FGFR3 as a novel therapeutic in lung adenocarcinoma.

Nanotechnology based anti-infectives to fight microbial intrusions.

With the rise in human population across the globe especially in developing countries, the incidence of microbial infections are increasing with greater pace. On the other hand, available medication and therapies are found to be insufficient for the complete cure of such microbial infections due to the development of resistance against various antibiotics. Therefore, to cope up the menace of microbial infections and drug resistance, there is demand for new and compelling technology, which has the ability to impede these problems. Many research groups worldwide are finding a ray of hope in nanomaterials owing to their unique properties. In the present review we have discussed the reasons behind the development of new materials based on nanotechnology. It is mainly focused on pioneering studies on application of nanomaterials like carbon nanotube, fullerene, dendrimers, nanocomposite and metal nanoparticles in combating dreadful pathogens. Moreover, the concerns about their toxicity have also been discussed.

Sequence analysis of Indian iris yellow spot virus ambisense genome segments: evidence of interspecies RNA recombination.

The nucleotide sequence of M- and S-RNA segments of an Indian iris yellow spot virus (IYSV) were determined. Sequence comparisons showed that both of these sequences shared less than 95 % identity with those other known IYSV isolates. Phylogenetic analysis revealed that the S- and M-RNA sequences of known IYSV isolates clustered with those of the tospoviruses, tomato yellow ring virus, polygonum ringspot virus and hippeastrum chlorotic ringspot virus. Further, multiple recombination detection methods detected inter- and intra-species recombination events that clustered primarily within the intergenic regions of S- and M-RNA, suggesting that these are possibly recombination hotspots in IYSV and closely related tospoviruses.

A preliminary Raman spectroscopic study of urine: diagnosis of breast cancer in animal models.

Prognosis of breast cancer, the most common cancer in females worldwide, has been shown to improve with early detection. Owing to disadvantages like low sensitivity, specificity, tedious sample preparation, long output times and inter-observer variance of currently available screening/diagnostic tools, rapid and objective alternatives such as Raman spectroscopy (RS) are being extensively explored. Body fluid (serum and saliva) based RS assays have shown promising results in diagnosis of oral, lung and nasopharyngeal cancers. The current study aims to explore the feasibility of breast cancer diagnosis using urine based RS. In this study, spectra were acquired from unprocessed as well as concentrated urine of controls (C) and breast tumor bearing (T) rats and analyzed using Principal Component Analysis (PCA) and Principal Component-Linear Discriminant Analysis (PC-LDA). Classification efficiencies of 80% and 72% using unprocessed urine and 78% and 91% using concentrated urine for C and T rats were achieved. Thus, results suggest the possibility of breast cancer diagnosis using urine based RS. Further, spectra were also acquired from concentrated urine samples collected prior to breast tumor development (TT) in rats and from rats that did not develop tumors despite carcinogen treatment (NTT). Concentrated urine of NTT rats could be classified as 'normal' (C or NTT) with ∼83% efficiency whereas concentrated urine from visibly and palpably normal rats that eventually developed tumor (TT rats) could be classified as 'abnormal' (TT or T) with ∼72.5% efficiency using PC-LDA. These results suggest the possibility of detecting biochemical changes occurring prior to tumor development using urine based RS.

Swiss bare mice: a suitable model for transcutaneous in vivo Raman spectroscopic studies of breast cancer.

Breast cancer is the most common cancer affecting females worldwide. As early detection results in better prognosis, screening tools for breast cancer are being explored. Raman spectroscopy, a rapid, objective, and noninvasive tool, has shown promising results in the diagnosis of several cancers including breast cancer. For development as a screening tool, a study of spectral signatures associated with breast cancer progression is imperative. However, such studies are not possible in human subjects. Hence, there is a need for a suitable animal model, which is conducive to transcutaneous in vivo Raman spectroscopic measurements of breast with minimal interference from skin and hair and has contribution from functional mammary epithelium of breast. In this study, rodent models like C57, Swiss albino, Swiss bare, agouti mice, and Sprague-Dawley rats were evaluated. Among these models, transcutaneous breast spectra of hairless Swiss bare mice have the best signal-to-noise ratio and were closest to reported ex vivo as well as intraoperative in vivo human breast spectra. Principal component-linear discriminant analysis of several anatomical sites confirms minimal skin interference and suggests contribution from functional mammary epithelium of breast. Moreover, transcutaneous spectra from normal breast and breast tumors of Swiss bare mice could be classified with 99% efficiency, which is better than the previous reports. Thus, Swiss bare mice model may be better suited for transcutaneous in vivo Raman spectroscopic studies of breast physiology and pathology, especially breast cancer. Prospectively, in addition to cancer progression, breast-to-bone metastasis can also be studied, since these anatomical sites can be uniquely classified.

Piecewise Linear Slope Estimation.

This paper presents a method for directly estimating slope values in a noisy piecewise linear function. By imposing a Markov structure on the sequence of slopes, piecewise linear fitting is posed as a maximum a posteriori estimation problem. A dynamic program efficiently solves this by traversing a linearly growing trellis. The alternating maximization algorithm (a kind of pseudo-EM method) is used to estimate the model parameters from data and its convergence behavior is analyzed. Ultrasound shear wave imaging is presented as a primary application. The algorithm is general enough for applicability in other fields, as suggested by an application to the estimation of shifts in financial interest rate data.

First Report of Iris yellow spot virus Infecting Allium tuberosum in India.

Allium tuberosum L., commonly known as garlic chives, is an important spice in northeastern India as well as in many other parts of the world. Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) is an important pathogen of onion (4) and other related Alliums such as garlic (3) and leek (2). During April 2013, symptoms potentially induced by IYSV such as chlorotic and straw-colored spindle-like lesions were observed on leaves of A. tuberosum accession Hanzong Winter (CGN 20779) plants in the wild species garden at the Directorate of Onion and Garlic Research (DOGR), Rajgurunagar, Pune, Maharashtra, India. Ten plant samples of A. tuberosum were randomly collected from the wild species garden and the upper, middle, and lower portions of the leaves were pooled and tested by double-antibody sandwich (DAS)-ELISA using a commercially available kit (Agdia Inc., Elkhart, IN) for IYSV. All of them showed positive results for IYSV incidence. Total RNA from the ELISA positive leaf samples of A. tuberosum was extracted using the RNeasy Plant Mini kit (Qiagen GmbH, Hilden, Germany). The primer pair IYSV-F (5'-TCAGAAATCGAGAAACTT-3') and IYSV-R (5'-TAATTATATCTATCTTTCTTGG-3') (1) was used for RT-PCR. The primer pair was specific to amplify 797 bp of the nucleocapsid (N) gene of IYSV. The amplified product derived from A. tuberosum isolate was purified by QIAquick PCR Purification Kit (Qiagen) and cloned using the vector pDrive (Qiagen). The recombinant clone was sequenced (Accession No. KF624624). Sequence analysis performed on CLC Main Workbench Version 6.8.4 confirmed that the fragment was of IYSV. Nucleotide sequence comparison of our virus with other IYSV isolates revealed that the highest nucleotide identity (99%) was with the IYSV garlic isolate (HM173691) from India. Further, maximum 96% protein identity was with IYSV onion isolate (ACA09432) and garlic isolate (ADK56108) from India. To our knowledge, this is the first report of IYSV naturally occurring on A. tuberosum in India. It is evident from previous studies that IYSV causes significant losses in onions (1) and from this study, that its symptoms have direct impact on quality of garlic chives. Further detailed studies are required to assess the magnitude of the impact of IYSV infection on yield and quality of A. tuberosum. References: (1) A. Bulajic et al. Plant Dis. 93:976, 2009. (2) M. C. Córdoba-Sellés et al. Plant Dis. 91:1365, 2007. (3) S. J. Gawande et al. Plant Dis. 94:1066, 2010. (4) B. Mandal et al. Plant Dis. 96:468, 2012.

First Report of Leek yellow stripe virus in Allium sativum in Western India.

Garlic (Allium sativum L.) is an important bulbous spice crop in India as well as other parts of world. Garlic is well known for its medicinal properties. Degeneration due to viral infections is one of the important constraints in exploiting its yield potential. Leek yellow stripe virus (LYSV), genus Potyvirus, family Potyviridae, is a prominent virus known to infect garlic worldwide (4). During July 2013, potyvirus-like symptoms such as mosaic, streaking, stunting, mottling of leaves were observed on garlic cv. G-41 and landrace Ranibennur local, collected from Karnataka, India, and maintained at the Directorate of Onion and Garlic Research, Rajgurunagar, Pune, India. The incidence of symptomatic plants was estimated at 70% for Ranibennur local and 68% for cv. G-41. The symptomatic leaves were sampled diagonally from the field. Twenty symptomatic plants per cultivar with each sample was composited from young, middle, and lower (basal) leaves of the plant. These samples were tested by double-antibody sandwich (DAS)-ELISA for LYSV using commercially available kit (Agdia Inc., Elkhart, IN). ELISA-positive plants were further subjected to molecular studies. Total RNA from the infected leaf samples were extracted by RNeasy Plant Mini kit (Qiagen GmbH, Hilden, Germany) and assayed by reverse transcription (RT)-PCR using primer pair LYSV-F 2 (5'-GCACCATACAGTGAATTGAG-3') (1), LYSV-R (5'-GCCTCGCGCGCTCTAA-3') (3) to amplify 874 bases of partial Nib and partial coat protein gene. The amplified product of 874 bp derived from A. sativum isolate was purified (QIAquick PCR Purification Kit, Qiagen) and cloned using vector pDrive (Qiagen). The recombinant clones were sequenced and submitted in NCBI database (GenBank Accession No. KF850539). The sequence analysis performed on CLC Main Workbench Version 6.8.4 gave confirmation of LYSV. Further, phylogenetic analysis of the 874-nt sequence revealed the highest nucleotide identity (80 to 82%) with LYSV isolates (DQ925453, JN127339, AB005611, and JX429965). To the best of our knowledge, this is the first report of natural infection of garlic by LYSV in western India. LYSV is known to cause direct losses in garlic and other related Allium spp. Up to 54% reduction in bulb weight was observed due to single infection of this virus (2). Hence, our first report about this virus has significant impact on garlic production scenario, if this virus found to be widespread in the country. For this, additional surveys and genotype screenings are needed to obtain a better understanding of the potential impact of LYSV on garlic production in India. References: (1) H. Fidan and S. Baloglu. Plant Dis. 93:672, 2009. (2) H. Lot et al. Plant Dis. 82:1381, 1998. (3) P. Lunello et al. J. Virol. Methods. 118:15, 2004. (4) H. R. Pappu et al. Plant Dis. 89:205, 2005.

The least-squares invertible constant-Q spectrogram and its application to phase vocoding.

This paper discusses the development of a constant-Q spectrogram representation that is invertible in a least-squares sense. A good quality inverse is possible because this modified transform method, unlike the usual sliding window constant-Q spectrogram, does not discard data samples when performing the variable length discrete Fourier transforms on the signal. The development of a phase vocoder application using this modified technique is also discussed. It is shown that a phase vocoder constructed using the least-squares invertible constant-Q spectrogram (LSICQS) is not a trivial extension of the regular FFT-based phase vocoder algorithm and some of the mathematical subtleties related to phase reassignment are addressed.

Silver nanoparticles: the powerful nanoweapon against multidrug-resistant bacteria.

In the present scenario, pharmaceutical and biomedical sectors are facing the challenges of continuous increase in the multidrug-resistant (MDR) human pathogenic microbes. Re-emergence of MDR microbes is facilitated by drug and/or antibiotic resistance, which is acquired way of microbes for their survival and multiplication in uncomfortable environments. MDR bacterial infections lead to significant increase in mortality, morbidity and cost of prolonged treatments. Therefore, development, modification or searching the antimicrobial compounds having bactericidal potential against MDR bacteria is a priority area of research. Silver in the form of various compounds and bhasmas have been used in Ayurveda to treat several bacterial infections since time immemorial. As several pathogenic bacteria are developing antibiotic resistance, silver nanoparticles are the new hope to treat them. This review discusses the bactericidal potential of silver nanoparticles against the MDR bacteria. This multiactional nanoweapon can be used for the treatment and prevention of drug-resistant microbes.

Novel laboratory mouse papillomavirus (MusPV) infection.

Most papillomaviruses (PVs) are oncogenic. There are at least 100 different human PVs and 65 nonhuman vertebrate hosts, including wild rodents, which have species-specific PV infections. Florid papillomatosis arose in a colony of NMRI-Foxn1(nu)/Foxn1(nu) (nude) mice at the Advanced Centre for Treatment Research and Education in Cancer in India. Lesions appeared at the mucocutaneous junctions of the nose and mouth. Histologically, lesions were classical papillomas with epidermal hyperplasia on thin fibrovascular stalks in a verrucous pattern. Koilocytotic cells were observed in the stratum granulosum of the papillomatous lesions. Immunohistochemically, these abnormal cells were positive for PV group-specific antigens. With transmission electron microscopy, virus particles were observed in crystalline intranuclear inclusions within keratinocytes. The presence of a mouse PV, designated MusPV, was confirmed by amplification of PV DNA with degenerative primers specific for PVs. This report is the first of a PV and its related disease in laboratory mice.

Phase I study of decitabine with doxorubicin and cyclophosphamide in children with neuroblastoma and other solid tumors: a Children's Oncology Group study.

BACKGROUND: Demethylating agents may alter the expression of genes involved in chemotherapy resistance. We conducted a phase I trial to determine the toxicity and molecular effects of the demethylating agent, decitabine, followed by doxorubicin and cyclophosphamide in children with refractory solid tumors. PROCEDURE: Stratum A included children with any solid tumor; Stratum B included neuroblastoma patients only. Patients received a 1-hr decitabine infusion for 7 days, followed by doxorubicin (45 mg/m(2)) and cyclophosphamide (1 g/m(2)) on day 7. Pharmacokinetic studies were performed after the first dose of decitabine. Biological studies included methylation and gene expression analyses of caspase-8, MAGE-1 and fetal hemoglobin (HbF), and expression profiling of pre- and post-treatment peripheral blood and bone marrow cells. RESULTS: The maximum-tolerated dose of decitabine was 5 mg/m(2)/day for 7 days. Dose-limiting toxicities at 10 mg/m(2)/day were neutropenia and thrombocytopenia. Decitabine exhibited rapid clearance from plasma. Three of 9 patients in Stratum A and 4/12 patients in Stratum B had stable disease for > or = 4 months. Sustained MAGE-1 demethylation and increased HbF expression were observed in the majority of patients post-treatment (12/20 and 14/16, respectively). Caspase-8 promoter demethylation and gene expression were seen in 2/7 bone marrow samples. Differentially expressed genes were identified by microarray analysis. CONCLUSION: Low-dose decitabine when combined with doxorubicin/cyclophosphamide has tolerable toxicity in children. However, doses of decitabine capable of producing clinically relevant biologic effects were not well tolerated with this combination. Alternative strategies of combining demethylating agents with non-cytotoxic, biologically targeted agents such as histone deacetylase inhibitors should be explored.

Fabrication of silver nanoparticles by Phoma glomerata and its combined effect against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus.

AIMS: We report extracellular synthesis of silver nanoparticles (Ag-NPs) from Phoma glomerata and its efficacy against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The bacteria exhibiting resistance to various antibiotics showed remarkable sensitivity, when used in combination of antibiotics and Ag-NPs. METHODS AND RESULTS: Biosynthesis of Ag-NPs was carried out by challenging the fungal cell filtrate with 1 mmol l(-1) silver nitrate. The Ag-NPs were characterized with the help of UV-Visible spectrophotometer and Fourier transform infrared spectroscopy. Scanning electron microscopy was carried out to detect the size of Ag-NPs. Evaluation of the combined effect(s) was studied by disc diffusion method against E. coli, Staph. aureus and Ps. aeruginosa. CONCLUSIONS: The biosynthesis route seems to be eco-friendly and easy to scale up the process. Thus, these Ag-NPs may prove as a better candidate for drugs and can potentially eliminate the problem of chemical agents because of their biogenic nature. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacterial resistance against antibiotics has been increasing with alarming rate. To overcome this problem, there is a pressing need to develop bactericidal agents. Ag-NPs may prove to be an answer to drug-resistant bacteria.

A simple and effective approach for the treatment of chronic wound infections caused by multiple antibiotic resistant Escherichia coli.

Antimicrobial resistance is a major problem in present-day therapy. Despite the advent of newer antimicrobial agents with a broad spectrum of activity, multiple antibiotic resistant pathogens are difficult to eliminate from infected sites. The present study was carried out to develop an approach, using citric acid as a sole antimicrobial agent, for the treatment of chronic wound infections caused by multiresistant Escherichia coli (MAREC). A total of 34 cases of chronic wound infections yielding MAREC isolates on culture were studied. The antibacterial effect of citric acid against MAREC was evaluated in vitro by broth dilution method. Three percent citric acid gel was applied to each wound once daily until it healed completely. All 34 isolates were inhibited by citric acid with minimum inhibitory concentrations in the range of 1500-2000 microg/ml. Topical application of 3% citric acid to wounds 7-42 times resulted in elimination of MAREC from infected sites and successful healing of wounds in all 34 patients. This treatment modality was simple, reliable, non-toxic and effective. Hence, the use of citric acid for the cost-effective treatment of wound infections caused by MAREC is recommended.

Mouse skin tumor promotion by sodium arsenate is associated with enhanced PCNA expression.

Drinking water contamination by arsenicals remains a major public health problem in many parts of the world more particularly in India and Bangladesh. Despite arsenic being a health hazard and implicated in human carcinogenesis, the experimental evidence available is much limited even now and the mechanisms involved during carcinogenesis and tumor promotions are not clear. Accordingly, in this study, we have studied the tumor promoter effects of sodium arsenate on mouse skin tumor promoter model system using 9,10-dimethyl-1,2-benzanthracene (DMBA) as a initiating carcinogen. Our studies showed development of papillomas on mice skin treated with only DMBA. However, mice treated with DMBA on skin and administered arsenate (As) in drinking water showed development of well differentiated squamous cell carcinomas. Further, both by immunohistochemistry and western blotting analysis studies higher levels of proliferating cell nuclear antigen (PCNA) was observed in mice treated with DMBA plus arsenate compared to only DMBA treated group. PCNA is known to be associated with S phase and DNA replication of the cell cycle. The plain controls and arsenate controls did not show significant difference either in tumor development or in PCNA levels. The present study demonstrates mouse skin tumor promoting effect of arsenate which seems to be associated with abnormal cell proliferation as indicated by higher levels of PCNA expression.

Bioevaluation of radioactive bandages in a murine model of melanoma.

PURPOSE: To study the effectiveness of a radioactive bandage incorporating a beta(-) emitter for the treatment of superficial tumours like melanoma. MATERIALS AND METHODS: (188)Re tin particles were immobilized on a bandage patch ((188)Re bandage). The effectiveness of the (188)Re bandage for controlling tumour growth was tested in C57BL/6 mice bearing BL6/FIO melanoma. The effect of the single dose delivered, two-dose treatment and time of contact of bandages on the skin was studied by following tumour size. RESULTS: Tumour growth was delayed significantly in treated animals compared with controls. Complete tumour regression was observed with some doses of radiation. Histology studies and dose-rate calculations were also carried out to evaluate the effectiveness of (188)Re bandages. CONCLUSIONS: Radioactive bandages could be a promising modality for the treatment of skin cancers.