The first record of Heterakis gallinarum as a cause of fatal nodular typhlitis in golden pheasants (Chrysolophus pictus) in India.

Background: Heterakidosis is one of the most prevalent parasitic diseases in birds, the caecae of a variety of wild and domestic birds are infected with these nematodes. In pheasants, nodular typhlitis is a lethal disease caused mainly by infection with Heterakis isolonche alone or in conjunction with Heterakis gallinarum. H. gallinarum has long been recognized to infect birds with low pathogenicity, with only a few fatal cases previously reported. Case description: This paper describes a case of fatal nodular typhlitis due to H. gallinarum in a male and female pair of adult golden pheasants (Chrysolophus pictus) from a zoological garden in Uttar Pradesh, India. Findings/treatment and outcome: The caecum had multiple serosal and mucosal nodules, the majority of which were found to contain various stages of parasites embedded in the center along with the free forms in the caecal contents. Histopathologically, these nodules were generally represented by granulomas centered on necrotic parasite debris, with the occasional reactive fibrous hyperplastic tissue reaction. Based on the morphology and nematode-specific internal transcribed spacer (ITS) ITS1-5.8 rRNA-ITS2 region-based PCR, the nematode was identified as H. gallinarum. The presence of H. gallinarum was further confirmed by sequencing the ITS region followed by phylogenetic analysis. According to the author's best knowledge, this is the first instance of H. gallinarum being linked to nodular typhlitis in pheasants in India. Conclusion: Our findings confirm that H. gallinarum, other than H. isolonche, can induce severe nodular typhlitis with a fatal outcome in pheasants.

Earth-abundant and environmentally benign Ni-Zn iron oxide intercalated in a polyaniline based nanohybrid as an ultrafast photodetector.

Nickel-zinc iron oxide (NZF) was introduced into a polyaniline (PANI) matrix by an in situ chemical oxidation polymerization approach. The surface composition and chemical states were investigated by X-ray photoelectron spectroscopy (XPS), which revealed an Fe 2p spectrum with the two peak positions of Fe 2p3/2 and Fe 2p1/2 at 711.00 and 724.48 eV, respectively. Deconvolution of the Fe 2p3/2 peak revealed two components with binding energies of 713.98 and 718.16 eV, corresponding to the presence of Fe cations in the octahedral and tetrahedral sites. Additionally, the Rietveld refinement of NZF showed a cubic system with the Fd3m space group. High-resolution transmission electron microscopy (HRTEM) analysis showed that the NZF material strongly interacts with polyaniline, while the selected area electron diffraction (SAED) pattern perfectly matched with the XRD data. Lognormal distribution was used to determine the particle size, which was found to be in the range of 1-100 nm. A flexible photodetector device utilizing the NZF-PANI nanohybrid was fabricated on an environmentally friendly, biodegradable cellulose paper substrate and the device exhibited excellent performance, i.e., a responsivity of 0.069 A W-1 and detectivity of 7.258 × 1010 Jones at a very low voltage of 0.1 V. The non-stretched device showed a responsivity of 24.980 A W-1 at 5 V, whereas at 2 cm-1 bending curvature, the device showed a responsivity of 20.175 A W-1, which was much higher than the responsivity of a commercial photodetector (<0.5 A W-1).

Risk factors and predictors of mortality in acute invasive fungal sinusitis - a single-institution experience.

OBJECTIVE: To determine risk factors affecting mortality in acute invasive fungal sinusitis. METHOD: This observational cohort study was conducted over a five-year period. RESULTS: Of 109 recruited patients, 90 (82.6 per cent) had diabetes mellitus. Predominant fungi were zygomycetes (72.6 per cent) with Rhizopus arrhizus being most common. Of the patients, 12.8 per cent showed a positive biopsy report from radiologically normal sinuses. Factors affecting mortality on multivariate analysis were: female sex (p = 0.022), less than two weeks between symptoms and first intervention (p = 0.01), and intracranial involvement (p = 0.034). Other factors significant on univariate analysis were: peri-orbital swelling (p = 0.016), restricted ocular movements (p = 0.053), intracranial symptoms (p = 0.008), posterior disease (p = 0.058), imaging showing ocular involvement (p = 0.041), fungus being zygomycetes (p = 0.050) and post-operative cavity infection (p = 0.032). Bilateral, palatal and retromaxillary involvement were not associated with poor prognosis. CONCLUSION: Diagnosis of acute invasive fungal sinusitis requires a high index of clinical suspicion. Recognition of factors associated with poor prognosis can help when counselling patients, and can help initiate urgent intervention by debridement and antifungal therapy. Post-operative nasal and sinus cavity care is important to reduce mortality.

Effect of prophylactic granulocyte-colony stimulating factor (G-CSF) on acute hematological toxicity in medulloblastoma patients during craniospinal irradiation (CSI).

OBJECTIVES: Haematological toxicity and treatment breaks are common during cranio-spinal irradiation (CSI) due to irradiation of large volume of bone marrow. We conducted this study to see the effect of prophylactic granulocyte colony stimulating factor (GCSF) in reducing treatment breaks. PATIENTS AND METHODS: The study was conducted over a period of 15 months from August 2017 to November 2018. Histopathologically proven Medulloblastoma patients received prophylactic GCSF during CSI. Acute hematological toxicities and treatment breaks were noted and effect of age and pretreatment blood counts were analyzed by SPSS (Statistical Package for Social Sciences) version 23. RESULTS: A total of 28 patients were included in the study. During CSI, hematological toxicity leading to treatment breaks was observed in 11 (39.3 %) patients, of which grade 3 and 2 toxicities were seen in ten and one patients respectively. Younger age (<10 years) at diagnosis was significantly associated with the development of hematological toxicity (p = 0.028, Chi-Square). No correlation was found with pre-treatment blood counts. CONCLUSION: Prophylactic use of GCSF may be effective in preventing radiation induced hematological toxicity and treatment breaks.

Nanostructured cobalt antimonate: a fast responsive and highly stable sensing material for liquefied petroleum gas detection at room temperature.

Herein, cobalt antimonate (CoSb2O6) nanospheres were fabricated via the sol-gel spin-coating process and employed as a functional liquefied petroleum gas (LPG) sensor at room temperature (25 °C). The microstructure of the fabricated CoSb2O6 thin films (thickness ∼ 250 nm) was analyzed via scanning electron microscopy, which revealed the growth of nanospheres having an average diameter of ∼45 nm. The XRD analysis demonstrated the crystalline nature of CoSb2O6 with a crystallite size of ∼27 nm. Finally, the fabricated thin films were investigated as sensors for LPG and carbon dioxide (CO2) at room temperature (25 °C) and 55% R.H. (relative humidity) with different concentrations in the range of 1000-5000 ppm. The sensing results demonstrated greater variations in the electrical properties of films for the incoming LPG than that of the CO2 gas adsorption. Furthermore, to ensure the long-term stability of fabricated sensors, they were tested periodically at 10 days interval, spanning a total duration of 60 days. In summary, our fabricated LPG sensor displayed high sensitivity (1.96), repeatability, quick response time (21 s) and high long-term stability (99%). Therefore, CoSb2O6 nanospheres can be functionalized as a potential LPG-sensitive material characterized by high sensitivity, reliability and stability at room temperature.

Synthesis and biological function of Nickel and Copper nanoparticles.

Nickel and Copper nanoparticles were synthesized by simple chemical method and studied for antimicrobial activities. The size of synthesized Nickel and Copper nanoparticles was found to be 24.00 nm and 13.13 nm respectively. The XRD analysis reveals the crystal system of Nickel and Copper nanoparticles. Copper nanoparticles were found orthorhombic whereas the nickel nanoparticles were monoclinic. The antimicrobial activities of Nickel and Copper nanoparticles dispersed in DMSO was investigated. Comparative sensitivity test of these synthesized nanoparticles was carried out against three pathogenic micro-organisms (Gram negative bacteria), viz. Escherichia coli, Klebsiella pneumoniae and Pneumonia Typhus, using agar diffusion cup plate method. Copper and Nickel nanoparticles have shown appreciable sensitivity at 100 µg/ml against all test micro-organisms. Comparatively, Copper nanoparticles were found to exhibit higher zone of inhibition (ZOI) than Nickel nanoparticles.

Protein functionalised self assembled monolayer based biosensor for colon cancer detection.

We report results of the studies relating to the fabrication of a surface plasmon resonance (SPR) based label-free immunosensor for real-time monitoring of endothelin-1 (ET-1), a colon cancer biomarker. A gold disk modified with a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid (11-MUA) was functionalised via covalent immobilization of monoclonal anti-ET-1 antibodies using EDC-NHS (1-(3-(dimethylamine)-propyl)-3-ethylcarbodiimide hydrochloride, N-hydroxy succinimide) chemistry. This immunosensing platform (ethanolamine/anti-ET-1/11-MUA/Au) was characterized via atomic force microscopy (AFM), contact angle (CA) and Fourier transform infrared (FT-IR) spectroscopic techniques. The fabricated SPR electrode was further used to detect ET-1 in the broad concentration range 2-100 pg mL-1, with a detection limit of 0.30 pg mL-1 and remarkable sensitivity of 2.18 mo pg-1mL. The adsorption mechanism was studied using monophasic model and the values of association (ka) and dissociation (kd) constants for anti-ET-1 and ET-1 binding were calculated to be 4.4 ±â€¯0.4 × 105 M-1 s-1 and 2.04 ±â€¯0.0003 × 10-3 s-1, respectively. The results obtained via analysis of serum samples of colorectal cancer patients were found to be in good agreement with those obtained from enzyme-linked immunosorbent assay (ELISA) technique. Further, electrochemical studies were performed to prove the efficacy of the fabricated platform as a point of care device for the detection of ET-1.

Vitamin B12 immunoassay interference in a patient with multiple myeloma - troubleshooting in a two step reagent kit based on enhanced chemiluminescence testing.

Immunoassays are widely used for quantification of serum analytes however they are subjected to interference by endogenous antibodies. The laboratory procedures used to identify these endogenous antibodies is the demonstration of response to dilution or use of nonimmunoglobulin protein to block the interfering antibodies or the use of an alternate immunoassay. We report a clinical-diagnostic situation where serum vitamin B12 determination was interfered in an immunoassay due to excess of endogenous antibodies from monoclonal gammopathy that resulted in excess of analyte concentration. Reporting of such cases may be beneficial when assaying sera of multiple myeloma to avoid false results and in addition to avoid costs due to unnecessary repeat testing and further delay reporting of results.

Actionable perturbations of damage responses by TCL1/ATM and epigenetic lesions form the basis of T-PLL.

T-cell prolymphocytic leukemia (T-PLL) is a rare and poor-prognostic mature T-cell malignancy. Here we integrated large-scale profiling data of alterations in gene expression, allelic copy number (CN), and nucleotide sequences in 111 well-characterized patients. Besides prominent signatures of T-cell activation and prevalent clonal variants, we also identify novel hot-spots for CN variability, fusion molecules, alternative transcripts, and progression-associated dynamics. The overall lesional spectrum of T-PLL is mainly annotated to axes of DNA damage responses, T-cell receptor/cytokine signaling, and histone modulation. We formulate a multi-dimensional model of T-PLL pathogenesis centered around a unique combination of TCL1 overexpression with damaging ATM aberrations as initiating core lesions. The effects imposed by TCL1 cooperate with compromised ATM toward a leukemogenic phenotype of impaired DNA damage processing. Dysfunctional ATM appears inefficient in alleviating elevated redox burdens and telomere attrition and in evoking a p53-dependent apoptotic response to genotoxic insults. As non-genotoxic strategies, synergistic combinations of p53 reactivators and deacetylase inhibitors reinstate such cell death execution.

Discovery of novel drug sensitivities in T-PLL by high-throughput ex vivo drug testing and mutation profiling.

T-cell prolymphocytic leukemia (T-PLL) is a rare and aggressive neoplasm of mature T-cells with an urgent need for rationally designed therapies to address its notoriously chemo-refractory behavior. The median survival of T-PLL patients is <2 years and clinical trials are difficult to execute. Here we systematically explored the diversity of drug responses in T-PLL patient samples using an ex vivo drug sensitivity and resistance testing platform and correlated the findings with somatic mutations and gene expression profiles. Intriguingly, all T-PLL samples were sensitive to the cyclin-dependent kinase inhibitor SNS-032, which overcame stromal-cell-mediated protection and elicited robust p53-activation and apoptosis. Across all patients, the most effective classes of compounds were histone deacetylase, phosphoinositide-3 kinase/AKT/mammalian target of rapamycin, heat-shock protein 90 and BH3-family protein inhibitors as well as p53 activators, indicating previously unexplored, novel targeted approaches for treating T-PLL. Although Janus-activated kinase-signal transducer and activator of transcription factor (JAK-STAT) pathway mutations were common in T-PLL (71% of patients), JAK-STAT inhibitor responses were not directly linked to those or other T-PLL-specific lesions. Overall, we found that genetic markers do not readily translate into novel effective therapeutic vulnerabilities. In conclusion, novel classes of compounds with high efficacy in T-PLL were discovered with the comprehensive ex vivo drug screening platform warranting further studies of synergisms and clinical testing.

Reporting of Medical Errors in Autopsied Cases.

This report describes the cases autopsied at B. P. Koirala Institute of Health Sciences- Nepal, during the period of two years in which the treatment provided falls below the accepted standard practice causing the death of the patient. Postmortem examination provides an opportunity to find out the actual cause of death in which the cause of death is suspicious. Sometimes the cause of death found through postmortem examination is different from the clinical cause of death. The autopsy finding on the cause of death will help doctors even to think about unusual conditions that lead to death and hence apply preventive measures for those unusual conditions. The cause of medical error is not always due to the doctor's mistake, but sometimes also due to the obsolete hospital protocol. The autopsy reporting of medical errors can be used to identify lacunae in hospital protocols, problem-prone clinical processes and suggest interventions that may reduce negligence.

Enhanced sensitivity to glucocorticoids in cytarabine-resistant AML.

We sought to identify drugs that could counteract cytarabine resistance in acute myeloid leukemia (AML) by generating eight resistant variants from MOLM-13 and SHI-1 AML cell lines by long-term drug treatment. These cells were compared with 66 ex vivo chemorefractory samples from cytarabine-treated AML patients. The models and patient cells were subjected to genomic and transcriptomic profiling and high-throughput testing with 250 emerging and clinical oncology compounds. Genomic profiling uncovered deletion of the deoxycytidine kinase (DCK) gene in both MOLM-13- and SHI-1-derived cytarabine-resistant variants and in an AML patient sample. Cytarabine-resistant SHI-1 variants and a subset of chemorefractory AML patient samples showed increased sensitivity to glucocorticoids that are often used in treatment of lymphoid leukemia but not AML. Paired samples taken from AML patients before treatment and at relapse also showed acquisition of glucocorticoid sensitivity. Enhanced glucocorticoid sensitivity was only seen in AML patient samples that were negative for the FLT3 mutation (P=0.0006). Our study shows that development of cytarabine resistance is associated with increased sensitivity to glucocorticoids in a subset of AML, suggesting a new therapeutic strategy that should be explored in a clinical trial of chemorefractory AML patients carrying wild-type FLT3.

HOX gene expression predicts response to BCL-2 inhibition in acute myeloid leukemia.

Inhibitors of B-cell lymphoma-2 (BCL-2) such as venetoclax (ABT-199) and navitoclax (ABT-263) are clinically explored in several cancer types, including acute myeloid leukemia (AML), to selectively induce apoptosis in cancer cells. To identify robust biomarkers for BCL-2 inhibitor sensitivity, we evaluated the ex vivo sensitivity of fresh leukemic cells from 73 diagnosed and relapsed/refractory AML patients, and then comprehensively assessed whether the responses correlated to specific mutations or gene expression signatures. Compared with samples from healthy donor controls (nonsensitive) and chronic lymphocytic leukemia (CLL) patients (highly sensitive), AML samples exhibited variable responses to BCL-2 inhibition. Strongest CLL-like responses were observed in 15% of the AML patient samples, whereas 32% were resistant, and the remaining exhibited intermediate responses to venetoclax. BCL-2 inhibitor sensitivity was associated with genetic aberrations in chromatin modifiers, WT1 and IDH1/IDH2. A striking selective overexpression of specific HOXA and HOXB gene transcripts were detected in highly BCL-2 inhibitor sensitive samples. Ex vivo responses to venetoclax showed significant inverse correlation to ß2-microglobulin expression and to a lesser degree to BCL-XL and BAX expression. As new therapy options for AML are urgently needed, the specific HOX gene expression pattern can potentially be used as a biomarker to identify venetoclax-sensitive AML patients for clinical trials.

Mandibular Reconstruction: Overview.

INTRODUCTION: Mandibular reconstruction has changed significantly over the years and continues to evolve with the introduction of newer technologies and techniques. PURPOSE: This article reviews the history of oromandibular reconstruction, biomechanics of mandible, summarizes the reconstruction options available for mandible with defect classification, goals in reconstruction, the various donor sites, current reconstructive options, dental rehabilitation and persistent associated problems. SUMMARY: Oromandibular reconstruction, although a challenge for the head and neck reconstructive surgeon, is now reliable and highly successful with excellent long-term functional and aesthetic outcomes with the use of autogenous bone grafts and current reconstructive options. The ideal reconstruction would provide a solid arch to articulate with the upper jaw, restoring swallowing speech, mastication, and esthetics. Autogenous vascularized bone grafts in combination with microsurgical techniques have revolutionized mandibular reconstruction in oral cancer surgery. Current trends in mandibular reconstruction aim to achieve reestablishment of a viable mandible of proper form and maxillary mandibular relationship while decreasing the need for invasive autogenous graft procurement. However the optimal reconstruction of mandibular defects is still controversial in regards to reconstructive options which include the donor site selection, timing of surgery and method of reconstruction.

Versatility of Pleuripotent Undifferentiated Stem Cells Aspirated from Bone Marrow and its Applications in Oral and Maxillofacial Surgery.

INTRODUCTION: Bone marrow (BM) derived pleuripotent undifferentiated stem cells represent a promising population for supporting new concepts in cellular therapy. AIM: The aim of this study is to evaluate the versatility of pleuripotent undifferentiated stem cells derived from BM aspiration and its applications in oral and maxillofacial surgical procedures. MATERIALS AND METHODS: A total of 30 patients out of which 15 were with hard tissue defects (cystic lesions n = 6, post surgical alveolar defects n = 4, peri implant defects n = 3, alveolar clefts n = 2) and 15 soft tissue lesions (leukoplakia and lichen planus n = 6, oral submucous fibrosis n = 7, post traumatic soft tissue loss n = 2) were included in the study on randomized clinical basis. The patients received autologous BM derived mononuclear cells which were being locally delivered into the lesion and followed up. The parameters used were (1) To compare and evaluate the bone regeneration by radiographic assessment at the end of 3rd and 6th month postoperatively. (2) Duration of the procedure. (3) Clinical improvement in the management of soft tissue lesions. (4) Assessment of wound healing by Vancouver burn scar assessment of wound. (5) Safety, postoperative infections and complications. RESULTS: For hard tissue lesions CT scans and OPG revealed adequate regenerated bone, bridging the defect after 3 months. Hounsfield units of regenerated bone after 6 months were more or less similar to native bone which was statistically significant (unpaired t test = p < 0.05). For soft tissue lesions (1) 7 cases of OSMF showed adequate clinical mouth opening (one way anova test = p < 0.05), reduction in burning sensation and blanching of mucosa, (2) 6 cases of leukoplakia and lichen planus and 2 cases of post traumatic soft tissue defects showed good clinical improvement by Vancouver burn scar assessment of wound index. CONCLUSION: The study shows that there is a definite beneficial effect in bone regeneration and soft tissue wound healing with the use of BM-derived mononuclear cells.

Novel drug candidates for blast phase chronic myeloid leukemia from high-throughput drug sensitivity and resistance testing.

Chronic myeloid leukemia in blast crisis (CML BC) remains a challenging disease to treat despite the introduction and advances in tyrosine kinase inhibitor (TKI) therapy. In this study we set out to identify novel candidate drugs for CML BC by using an unbiased high-throughput drug testing platform. We used three CML cell lines representing different types of CML blast phases (K562, EM-2 and MOLM-1) and primary leukemic cells from three CML BC patients. Profiling of drug responses was performed with a drug sensitivity and resistance testing platform comprising 295 anticancer agents. Overall, drug sensitivity scores and the drug response profiles of cell line and primary cell samples correlated well and were distinct from other types of leukemia samples. The cell lines were highly sensitive to TKIs and the clinically TKI-resistant patient samples were also resistant ex vivo. Comparison of cell line and patient sample data identified new candidate drugs for CML BC, such as vascular endothelial growth factor receptor and nicotinamide phosphoribosyltransferase inhibitors. Our results indicate that these drugs in particular warrant further evaluation by analyzing a larger set of primary patient samples. The results also pave way for designing rational combination therapies.

The effects of social defeat on behavior and dopaminergic markers in mice.

The present study investigated the effects of chronic social defeat stress on several behavioral parameters, and the expression of dopaminergic markers, i.e., dopamine D1 receptors (D1Rs), dopamine D2 receptors (D2Rs), and dopamine and cyclic adenosine 3',5'-monophosphate-regulated phosphoprotein-32 (DARPP-32), in the prefrontal cortex (PFC), amygdala (AMY), and hippocampus (HIP) of mouse brains. After 10days of social defeat stress, the defeated mice were divided into two groups: one group underwent a series of behavioral tests. The other group was sacrificed on the 11th day and tissue samples were collected for Western blotting. The behavioral tests comprised tests of locomotion, light/dark preference, social interaction, as well as the novel object recognition test (NORT), Morris water maze, and forced swimming test (FST). We measured the expression of D1Rs, D2Rs, total DARPP-32, phospho-Thr34 or Thr75-DARPP-32 using Western blotting. The defeated mice showed increased anxiety- and depression-like behaviors, and impaired cognition. No significant differences in D1Rs and D2Rs expression were shown between defeated and control mice in any area studied. A significantly increased expression in total DARPP-32, and phospho-DARPP-32 was observed in the PFC or AMY of defeated mice. These data suggest that alterations in dopaminergic markers may be involved in anxiety- and depression-like behaviors, and cognitive impairment induced by social defeat stress.

Suppression of BRCA1 sensitizes cells to proteasome inhibitors.

BRCA1 is a multifunctional protein best known for its role in DNA repair and association with breast and ovarian cancers. To uncover novel biologically significant molecular functions of BRCA1, we tested a panel of 198 approved and experimental drugs to inhibit growth of MDA-MB-231 breast cancer cells depleted for BRCA1 by siRNA. 26S proteasome inhibitors bortezomib and carfilzomib emerged as a new class of selective BRCA1-targeting agents. The effect was confirmed in HeLa and U2OS cancer cell lines using two independent siRNAs, and in mouse embryonic stem (ES) cells with inducible deletion of Brca1. Bortezomib treatment did not cause any increase in nuclear foci containing phosphorylated histone H2AX, and knockdown of BRCA2 did not entail sensitivity to bortezomib, suggesting that the DNA repair function of BRCA1 may not be directly involved. We found that a toxic effect of bortezomib on BRCA1-depleted cells is mostly due to deregulated cell cycle checkpoints mediated by RB1-E2F pathway and 53BP1. Similar to BRCA1, depletion of RB1 also conferred sensitivity to bortezomib, whereas suppression of E2F1 or 53BP1 together with BRCA1 reduced induction of apoptosis after bortezomib treatment. A gene expression microarray study identified additional genes activated by bortezomib treatment only in the context of inactivation of BRCA1 including a critical involvement of the ERN1-mediated unfolded protein response. Our data indicate that BRCA1 has a novel molecular function affecting cell cycle checkpoints in a manner dependent on the 26S proteasome activity.

A pre-clinical model of resistance to induction therapy in pediatric acute lymphoblastic leukemia.

Relapse and acquired drug resistance in T-cell acute lymphoblastic leukemia (T-ALL) remains a significant clinical problem. This study was designed to establish a preclinical model of resistance to induction therapy in childhood T-ALL to examine the emergence of drug resistance and identify novel therapies. Patient-derived T-ALL xenografts in immune-deficient (non-obese diabetic/severe combined immunodeficient) mice were exposed to a four-drug combination of vincristine, dexamethasone (DEX), L-asparaginase and daunorubicin (VXLD). 'Relapse' xenografts were characterized by responses to drugs, changes in gene expression profiles and Connectivity Map (CMap) prediction of strategies to reverse drug resistance. Two of four xenografts developed ex vivo and in vivo drug resistance. Both resistant lines showed altered lipid and cholesterol metabolism, yet they had a distinct drug resistance pattern. CMap analyses reinforced these features, identifying the cholesterol pathway inhibitor simvastatin (SVT) as a potential therapy to overcome resistance. Combined ex vivo with DEX, SVT was significantly synergistic, yet when administered in vivo with VXLD it did not delay leukemia progression. Synergy of SVT with established chemotherapy may depend on higher drug doses than are tolerable in this model. Taken together, we have developed a clinically relevant in vivo model of T-ALL suitable to examine the emergence of drug resistance and to identify novel therapies.

A New Begomovirus Species in Association with Betasatellite Causing Tomato Leaf Curl Disease in Gandhinagar, India.

In December 2012, tomato leaf curl disease (ToLCD) (2) was observed in tomato-growing areas of Gandhinagar District of Gujarat, a state in northwestern India. Incidence of ToLCD was estimated to be between 40 and 70% depending on the cultivars used. Infected plants exhibited symptoms consisting of leaf rolling, leaf curling, and yellowing typical of begomoviruses. Total DNA was isolated from a single affected tomato plant (2). Begomovirus infection in this sample was established by amplification of the expected-size 550-bp DNA fragment from this extract by PCR with degenerate DNA-A primers (3). Rolling circle amplification (RCA) using ϕ29 DNA polymerase was carried out on the total DNA, followed by digestion with Bam HI. An amplicon of ~2.8 kb was gel-eluted and cloned into Bam HI linearized pBluescript II KS(+). Restriction enzyme digestion of plasmid DNA from the resulting clones indicated the presence of one type of molecule. Using PCR and universal betasatellite primers, the expected 1.3-kb fragment was amplified from the DNA extract (1). An amplicon of ~1.3 kb was gel-eluted and cloned into pTZ57RT vector. Sequence analysis revealed that DNA-A (GenBank Accession No. KC952005) is composed of 2,753 nt and showed the highest identity (87.8%) with Tomato leaf curl Kerala virus[India:Kerala:2008] (GenBank Accession No. EU910141). An analysis for recombination showed this begomovirus DNA likely to have originated by recombination between Tomato leaf curl Kerala virus and Tomato leaf curl Karnataka virus. The satellite DNA-ß (GenBank Accession No. KC952006) is composed of 1,365 nt and showed the highest identity (75.6%) with Tomato leaf curl betasatellite[India:Ludhiana:2004] (ToLCB-[IN:Lud:04]) (GenBank Accession No. AY765255). On the basis of DNA-A sequence analysis, the ICTV species demarcation criteria of 89% DNA-A sequence identity, and genome organization, the present isolate was considered as a new begomovirus species and named Tomato leaf curl Gandhinagar virus (ToLCGNV). The betasatellite shares less than 78% identity with (ToLCB-[IN:Lud:04]), it is considered a new species of betasatellite and the name, Tomato leaf curl Gandhinagar betasatellite (ToLCGNB) is proposed. Multimeric clones of the begomovirus and betasatellite DNAs were generated in a binary vector and these plasmids transformed into Agrobacterium tumefaciens. Nicotiana benthamiana and tomato plants agroinoculated with the cloned begomovirus DNA developed leaf curl symptoms, whereas plants co-agroinoculated with the cloned begomovirus and betasatellites developed more severe symptoms, including leaf rolling, leaf curling, and yellowing. The symptoms induced by the begomovirus and betasatellite DNAs were indistinguishable from those observed in the field. Thus, ToLCGNV is a new monopartite begomovirus which, in association with a new species of betasatellite, causes ToLCD in Gandhinagar, India. The presence of ToLCGNV needs to be considered, along with the already reported begomoviruses infecting tomatoes in this state, e.g., Tomato leaf curl Gujarat virus (2), in studies aimed to developing tomato cultivars with stable resistance to these tomato-infecting begomoviruses in India. References: (1) R. W. Briddon et al. Mol. Biotechnol. 20:315, 2002. (2) C. Reddy et al. Arch Virol. 150:845, 2005. (3) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.