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The present review deals with the serological and molecular techniques available for detection of fungal pathogens in plants, followed by management practices using bioinoculants which can induce resistance in the hosts. Specific detection of pathogens in the plant tissues even before the appearance of symptoms are done either through protein based immunodiagnostic techniques or DNA/RNA based molecular techniques. Immunotechniques depend on specific interaction between antigen of the pathogen or infected tissues and antibodies prepared against the specific antigen. Positive reactions are detected through either qualitative or quantitative techniques. Among the immunodiagnostic techniques, the most widely and commonly used is the enzyme linked immunosorbent assay (ELISA), Dot blot, Western blot and Immunofluorescence. The molecular techniques are predominantly PCR based and include different variations with increasing sensitivity such as nested PCR, multiplex PCR, reverse transcriptase PCR, real time PCR, loop-mediated isothermal amplification (LAMP) and nano based PCR. Thus, by use of one or more sophisticated techniques it is possible to detect pathogens rapidly and specifically. Pathogen detection is generally followed by management techniques, and in the current scenario ecologically safe bioinoculants are used for induction of immunity in the plants. There are several bioinoculants based on beneficial microorganisms such as plant growth promoting bacteria, biocontrol agents and arbuscular mycorrhizal fungi. Application of these microorganisms, singly or as a consortium, has been reported to induce immunity in plants. Induced systemic resistance (ISR) operates through signal molecules and pathways involving jasmonate and ethylene, and in very few cases, salicylate. Upregulation of these pathways leading to enhanced accumulation of antimicrobial compounds (phytoalexin) help in warding off the pathogen during subsequent attack.

The COVID 19 global pandemic has engulfed humanity with a huge impact on health systems across the world. Many patients develop myocardial injury which can lead to significant cardiovascular complications including HF. This will require aggressive management strategies which are evolving. Guideline directed drug therapy including ACEI/ARB/ARNI is to be continued in patients with pre-existing HF. Long-term cardiovascular effects of COVID-19 are yet to be ascertained. Protection of health care personnel from contracting the disease should be given high priority.

The unprecedented and rapidly spreading Coronavirus Disease-19 (COVID-19) pandemic has challenged public health care systems globally. Based on worldwide experience, India has initiated a nationwide lockdown to prevent the exponential surge of cases. During COVID-19, management of cardiovascular emergencies like acute Myocardial Infarction (MI) may be compromised. Cardiological Society of India (CSI) has ventured in this moment of crisis to evolve a consensus document for care of acute MI. However, this care should be individualized, based on local expertise and governmental advisories.

Background Utilization of cardiac Magnetic Resonance Imaging (cMRI) as an imaging modality in clinical practice is rapidly increasing. More evidence from randomized studies establishing clinical safety and performance of pacing systems in patients undergoing a cMRI scan is needed. Objectives The purpose of this prospective, multi-center, randomized study was to demonstrate safety and efficacy of the Accent MRI conditional pacing systems in patients undergoing cMRI scan. Methods Patients (n = 283) indicated for dual-chamber pacemaker implant were randomized to either the MRI Group (MG) (n = 140) or the Control Group (CG) (n = 143) after successful device implantation of the Accent MRI system. Clinical evaluation and device interrogation were performed at pre- and post-MRI scan, and one month post-MRI for all patients. At 9–12 weeks post-implant, patients in MG underwent a non-diagnostic cMRI scan at 1.5 Tesla (T) while patients in CG underwent device interrogation and clinical evaluation twice with a 45-minute waiting period in between. The safety endpoint was freedom from MRI scan-related complications and that for efficacy was significant changes in right atrial/ventricular (RA/RV) capture threshold and sensing amplitude between right before MRI, immediately after MRI and one month post-MRI. Results Results showed 100% freedom from MRI scan related complications. There were no significant changes in device performance between pre-MRI scan and one month post MRI scan time points in both study groups. Conclusions cMRI scanning with 1.5 T scanners is safe in patients implanted with the Accent MRI conditional pacing system and has no significant effect on the electrical parameters of the device and leads. This article is protected by copyright. All rights reserved

Key Points Question Can a technology-enabled, dynamically adaptive protocol be efficiently used to provide early and accurate detection of toxic effects to immune checkpoint inhibitors? Findings In this cohort study including 47 individuals with genitourinary cancers, a median patient adherence rate of 74% and a care team–automated alert review rate of 73% within 3 days without incurring the cost of increasing care team staffing was found. Dizziness, nausea and vomiting, and shortness of breath were the symptoms with the highest positive predictive value for adverse events requiring acute interventions. Meaning The findings of this study suggest that technology-enabled monitoring of patient-reported outcomes may provide a useful model for delivering complex care remotely in patients receiving immune checkpoint inhibitors., Importance Immune checkpoint inhibitors can produce distinct toxic effects that require prompt recognition and timely management. Objective To develop a technology-enabled, dynamically adaptive protocol that can provide the accurate information needed to inform specific remedies for immune toxic effects in patients treated with immune checkpoint inhibitors. Design, Setting, and Participants An open-label cohort study was conducted at a single tertiary referral center from September 6, 2019, to September 3, 2020. The median follow-up duration was 63 (interquartile range, 35.5-122) days. Fifty patients with genitourinary cancers treated with immune checkpoint inhibitors were enrolled. Interventions A fit-for-purpose electronic platform was developed to enable active patient and care team participation. A smartphone application downloaded onto patients’ personal mobile devices prompted them to report their symptoms at least 3 times per week. The set of symptoms and associated queries were paired with alert thresholds for symptoms requiring clinical action. Main Outcomes and Measures The primary end point of this interim analysis was feasibility, as measured by patient and care team adherence, and lack of increase in care team staffing. Operating characteristics were estimated for each symptom alert and used to dynamically adapt the alert thresholds to ensure sensitivity while reducing unnecessary alerts. Results Of the 50 patients enrolled, 47 had at least 1 follow-up visit and were included in the analysis. Median age was 65 years (range, 37-86), 39 patients (83%) were men, and 39 patients (83%) had metastatic cancer, with the most common being urothelial cell carcinoma and renal cell carcinoma (22 [47%] patients each). After initial onboarding, no further care team training or additional care team staffing was required. Patients had a median study adherence rate of 74% (interquartile range, 60%-86%) and 73% of automated alerts were reviewed within 3 days by the clinic team. Symptoms with the highest positive predictive value for adverse events requiring acute intervention included dizziness (21%), nausea/vomiting (26%), and shortness of breath (14%). The symptoms most likely to result in unnecessary alerts were arthralgia and myalgia, fatigue, and cough. Conclusions and Relevance The findings of this cohort study suggest an acceptable and fiscally sound method can be developed to create a dynamic learning system to detect and manage immune-related toxic effects., This cohort study conducts an interim analysis of an electronic platform developed for patients to report symptoms possibly related to the toxic effects of immune checkpoint inhibitors.

Recent studies suggest that JAK2 serves as a novel therapeutic target in Bcr-Abl+ chronic myelogenous leukemia (CML). We have reported the existence of an HSP90- associated high molecular weight network complex (HMWNC) that is composed of HSP90 client proteins BCR-ABL, JAK2, and STAT3 in wild type Bcr-Abl+ leukemic cells. Here we showed that the HSP90-HMWNC is present in leukemia cells from CML patients in blast stage, and in Imatinib (IM)-resistant 32Dp210 (T315I) leukemia cells. We found that the HSP90-HMWNC could be disassembled by depleting JAK2 with either Jak2-specific shRNA or treatment with JAK2 inhibitors (TG101209 or Ruxolitinib) and HSP90 inhibitor (AUY922). Combinational treatment with JAK2 and HSP90 inhibitors diminished the activation of BCR-ABL, JAK2 and its downstream targets. As a result, the IM-resistant 32Dp210 T315I cells underwent apoptosis. When administered in mice bearing 32Dp210 T315I leukemia, combinational therapy using Ruxolitinib and AUY922 prolonged the survival significantly. Thus, a combination of JAK2 and HSP90 inhibitors could be a powerful strategy for the treatment of CML, especially in IM-resistant patients.

Community-based rehabilitation is the strategy endorsed by the World Health Organization and other international bodies to promote the inclusion of people with disabilities, particularly in low- and middle-income countries. In this article we trace the journey of Edawu, a mental health rehabilitation unit in a rural area of Benue State, Nigeria, from an in-patient rehabilitation unit to a community-focused service. The partnership of organisations from the UK with Edawu along the journey is also described. The authors set out learning points from the project and the principles behind sustainable overseas organisational partnerships.

Despite the success of imatinib mesylate (IM) in the early chronic phase of chronic myeloid leukemia (CML), patients are resistant to IM and other kinase inhibitors in the later stages of CML. Our findings indicate that inhibition of Janus kinase 2 (Jak2) in Bcr–Abl+ cells overcomes IM resistance although the precise mechanism of Jak2 action is unknown. Knocking down Jak2 in Bcr–Abl+ cells reduced levels of the Bcr–Abl protein and also the phosphorylation of Tyr177 of Bcr–Abl, and Jak2 overexpression rescued these knockdown effects. Treatment of Bcr–Abl+ cells with Jak2 inhibitors for 4–6 h but not with IM also reduced Bcr–Abl protein and pTyr177 levels. In vitro kinase experiments performed with recombinant Jak2 showed that Jak2 readily phosphorylated Tyr177 of Bcr–Abl (a Jak2 consensus site, YvnV) whereas c-Abl did not. Importantly, Jak2 inhibition decreased pTyr177 Bcr–Abl in immune complexes but did not reduce levels of Bcr–Abl, suggesting that the reduction of Bcr–Abl by Jak2 inhibition is a separate event from phosphorylation of Tyr177. Jak2 inhibition by chemical inhibitors (TG101209/WP1193) and Jak2 knockdown diminished the activation of Ras, PI-3 kinase pathways and reduced levels of pTyrSTAT5. These findings suggest that Bcr–Abl stability and oncogenic signaling in CML cells are under the control of Jak2.

Chronic myelogenous leukemia (CML) patients treated with imatinib mesylate (IM) become drug resistant by mutations within the kinase domain of Bcr–Abl, and by other changes that cause progression to advanced stage (blast crisis) and increased expression of the Lyn tyrosine kinase, the regulation of which is not understood yet. In Bcr–Abl+ cells inhibition of Jak2, a downstream target of Bcr–Abl, by either Jak2 inhibitors or Jak2-specific short interfering RNA (siRNA) reduced the level of the SET protein, and increased PP2A Ser/Thr phosphatase and Shp1 tyrosine phosphatase activities, which led to decreased levels of activated Lyn. Activation of PP2A combined with Jak2 inhibition enhanced the reduction of activated Lyn kinase compared with Jak2 inhibition alone. In contrast, inhibition of either PP2A or Shp1 combined with Jak2 inhibition interfered with the loss of Lyn kinase activation more so than Jak2 inhibition alone, indicating the involvement of PP2A and Shp1 in the inactivation of the Lyn kinase caused by Jak2 inhibition. Inhibition of Jak2 induced apoptosis and reduced colony formation in IM-sensitive and -resistant Bcr–Abl mutant cell lines. Jak2 inhibition also induced apoptosis in CML cells from blast crisis patients but not in normal hematopoietic cells. These results indicate that Lyn is downstream of Jak2, and Jak2 maintains activated Lyn kinase in CML through the SET–PP2A–Shp1 pathway.

National guidelines exist to help schools in supporting pupils with medical needs. However, school staff are not contractually bound to help with the administration of medication and policies differ across schools, depending on local needs and views. A postal survey was conducted in Sheffield to elicit schools’ views regarding medication administration. Over 80 percent of the responding schools were willing to administer long-term medication and 67 percent would help with short-term needs as well. Written information from professionals is valued by most schools and many would prefer that dosages of medication are adjusted to avoid school hours. Of the schools, 17 percent did not want to take any responsibility for medication administration. Reluctance seems to arise from perceived lack of training and fear of litigation. These issues need to be addressed in order to improve school staff’s confidence with the administration of medication.

Attack of tea plant (Camellia sinensis (L.) O. Kuntze) leaves by the tea mosquito bugHelopeltis theivora (Hemiptera: Miridae) was positively correlated to temperature and rainfall, and partially to humidity, as determined in 12 varieties during the period 2000–2002. The insect attack was severe during the months of May–September, which had high temperature and rainfall, and led to severe loss of biomass due to curling and drying up of the leaves. The biochemical response of these 12 varieties of tea to attack by the insect was determined with special reference to oxidative enzymes and flavonoid flavor components. Insect attack led to an increase in the activities of the oxidative enzymes peroxidase, ascorbate peroxidase and polyphenol oxidase. Activities of phenyl alanine ammonia lyase generally decreased as a result of insect attack, which was significant in the United Planters Association of South India (UPASI) varieties. A significant decrease in polyphenols was also obtained in UPASI varieties. HPLC analysis of catechins revealed a decrease in some of the catechins in the infected leaves. Analysis of theaflavins from infusion of healthy andHelopeltis-infested tea leaves revealed no changes.

Biomethanation is one of the desirable options for obtaining clean fuel from abundant renewable biomass resources. Improvement of biomethane production may be achieved by using improved strains of microbes, particularly the terminal microbes – the methanogens. Attempts have been made to improve the efficiency of the methanogens isolated from local sources by subjecting the methanogens to mutagenic changes by physical (by γ irradiation, neutron bombardment) or chemical (by addition of chemicals like acridine orange, colchicine) means. The effects of the treated methanogens on biomethanation were studied. γ Irradiation or neutron bombardment mutagenesis was dose-dependent and time-dependent. High doses proved to be lethal but methanogens were found to be to some extent radiation resistant when subjected to irradiation at small doses for short duration (∼5–10 s). No or marginal improvement of methane production occurred for the two strains TDM and TRM. Improvement of methane production occurred from successive transfers of radiation treated strain SSM. Chemical mutagens invariably inhibited biomethanation and the inhibition was dose dependent.

The effect of supplemental alpha tocopherol and its mode of action were evaluated during carcinogenesis in uterine cervix of mice by chronic exposure to 20-methylcholanthrene (MC). Oral administration of alpha tocopherol restricted the progression of dysplastic changes in cervix uteri which lead to carcinoma in situ during chronic exposure to MC. Enhanced mitogenic stimulation of peripheral blood lymphocytes (PBL) indicating improved immune response was also noted following treatment. A decrease in micronucleus frequency in the vaginal exfoliated cells suggested reduced genotoxic effect of the carcinogen after alpha tocopherol administration. A decrease in lipid peroxides following ingestion of alpha tocopherol imply its protective role on DNA damage by oxidative free radicals. Similarly, elevated glutathione level and increased activities of glutathione-S-transferase, glutathione peroxidase, catalase and superoxide dismutase indicated involvement of alpha tocopherol in host's detoxification system.

The chemopreventive action of quercetin was examined during 20-methyl cholanthrene induced cervical neoplasia in virgin Swiss albino mice. The effects were evaluated on the basis of histopathological observation of the cervical epithelium, micronucleus frequency in vaginal exfoliated cells and some biochemical parameters in the host liver. Quercetin was found to arrest or reverse the progression of cervical neoplasia. The micronucleus frequency was reduced following its administration. The potential anti-carcinogenic effect of quercetin noted in this study is attributed to its antioxidant property which was reflected in the lipid peroxides and their role in the host detoxification system, as expressed in liver glutathione level, glutathione-S-transferase, glutathione peroxidase, catalase and superoxide dismutase activity. As an integral part of the diet quercetin may offer protection to the epithelium from the damaging effects of carcinogenic chemicals.

Findings from this 1989 Bangladesh nationally representative Contraceptive Prevalence Survey reveal that the survival status of the index child was a strong factor in explaining differences in birth spacing. The effect of mortality of the index child at the infant stage was shortening of the birth interval. The effect was weaker for child mortality. The age of the mother at child birth had the next strongest effect on birth spacing. Increases in maternal age at birth were associated with increases in birth interval. Explanatory variables that had little influence on birth spacing were maternal educational status sex of the index child residence and birth order. The data pertain to open and closed birth intervals during 1984-89. There were 11034 birth intervals of which 36% were closed birth intervals. The analysis included the distribution of closed subsequent birth intervals among different subgroups and the study of the differences. The product-limit approach for estimating the survival function was used in the analysis of both mean closed and open birth intervals. About 75% of birth intervals among women aged under 35 years ranged 13-24 and 25-36 months. About 66% of birth intervals among women aged over 35 years lasted 13-35 months; 19-24% lasted over 3 years. About 25% of births occurred within 12 months after women experienced the death of an infant. Only 5% of births occurred within 12 months among women without a prior index child death. About 50% of women had a subsequent birth within 13-24 months after the death of the index child. The product-limit estimates confirmed that the death of an index child during infancy was a strong determinant of a short subsequent birth interval.

The aim of this study is to understand the effect of elevated temperature on wheat seedlings of four cultivars, Gayetri (GY), Gandhari (GN), Kedar (KD), PBW343 in terms of tolerance/susceptibility , along with antioxidative and biochemical responses. For this, seedlings of four cultivars were exposed to different temperatures ranging from 25-400C for six hours. According to heat susceptibility index calculated GN was found to be heat tolerant, PBW343 heat sensitive and the other two, GY and KD moderately tolerant. Exposure to high temperature led to gradual increase in membrane lipid peroxidation and hydrogen peroxide accumulation. H2O2 and malonaldehyde accumulated to much higher amount in leaf tissues of PBW343 and GY than GN and KD. Among antioxidative enzymes, catalase, glutathione reductase and ascorbate peroxidase activity initially increased at 350C, followed by reduced activity under heat stress in all cultivars. However, peroxidase activity continued to increase under heat stress. Total chlorophyll and non enzymatic antioxidant carotenoids, initially increased upto 300C, following which there was gradual decrease in these components. Compatible osmolytes - proline and total sugar level within cellular compartments were enhanced upto 350C. Results suggested that wheat can endure heat induced oxidative stress up to certain period, manifested by elevation of non enzymatic antioxidants and osmo protectants, as well as upregulation of antioxidative enzymes.

The government of Bangladesh since independence has tried to strengthen the national family planning program. Using data from the 1989 Bangladesh Fertility Survey on a nationally representative sample of 7984 rural and urban households the authors consider fifteen socioeconomic and demographic variables and assess their relative importance in relation to current contraceptive use. The data indicate that 31.1% of younger and lower parity women use contraceptives. Womens education followed by womens participation in family planning decision making were the most important of selected factors which positively affect current use of contraception. Administrative division desire for additional children urban residence and family planning worker visits were also important factors significantly and positively related to the current use of contraception. Husbands occupation was strongly associated with contraceptive use. Child loss however was negatively associated with contraceptive use followed by sex composition of living children. The authors suggest that women should be encouraged to participate in family decisions and become informed about their rights privileges and family law. Finally it was found that education makes less difference to contraceptive use where family planning programs are strong.

Bcr-Abl is the predominant therapeutic target in chronic myeloid leukemia (CML), and tyrosine kinase inhibitors (TKIs) that inhibit Bcr-Abl have been successful in treating CML. With progression of CML disease especially in blast crisis stage, cells from CML patients become resistant to imatinib mesylate (IM) and other TKIs, resulting in relapse. Because Bcr-Abl is known to drive multiple signaling pathways, the study of the regulation of stability of Bcr-Abl in IM-resistant CML cells is a critical issue as a possible therapeutic strategy. Here, we report that a new dual-kinase chemical inhibitor, ON044580, induced apoptosis of Bcr-Abl+ IM-sensitive, IM-resistant cells, including the gatekeeper Bcr-Abl mutant, T315I, and also cells from blast crisis patients. In addition, IM-resistant K562-R cells, cells from blast crisis CML patients, and all IM-resistant cell lines tested had reduced ability to form colonies in soft agar in the presence of 0.5 µM ON044580. In in vitro kinase assays, ON044580 inhibited the recombinant Jak2 and Abl kinase activities when the respective Jak2 and Abl peptides were used as substrates. Incubation of the Bcr-Abl+ cells with ON044580 rapidly reduced the levels of the Bcr-Abl protein and also reduced the expression of HSP90 and its client protein levels. Lysates of Bcr-Abl+ cell lines were found to contain a large signaling network complex composed of Bcr-Abl, Jak2, HSP90, and its client proteins as detected by a gel filtration column chromatography, which was rapidly disrupted by ON044580. Therefore, targeting Jak2 and Bcr-Abl kinases is an effective way to destabilize Bcr-Abl and its network complex, which leads to the onset of apoptosis in IM-sensitive and IM-resistant Bcr-Abl+ cells. This inhibitory strategy has potential to manage all types of drug-resistant CML cells, especially at the terminal blast crisis stage of CML, where TKIs are not clinically useful.

The role of a corrosive environment on polymeric targets during ion bombardment, has been observed. Presence of elastomeric seals or PVC etc. inside the vacuum chamber creates a partial pressure of HCl vapours. Observation of the decrease in the partial pressure of the HCl content as H+ ions impinge on a PVDF sample, indicates the possibility of attachments of HCl or Cl ions with the irradiated polymeric surface. This has been confirmed by residual gas analysis and by EDAX measurements of irradiated and unirradiated surfaces of PVDF samples.

A 63 year old woman presented in March 1997 with a three month history of shortness of breath while lying flat and in the left lateral position. She had a history of ischaemic heart disease and had coronary artery bypass surgery five years ago. In August 1995 she had an inferior wall myocardial infarction with postinfarction angina, which responded to medical treatment. Echocardiographic window was poor because of obesity; however, there appeared to be a large echo free space postero-inferior to the left ventricle leading to suspicion of left ventricular pseudoaneurysm. Magnetic resonance images of the heart performed in a …

The numbers of argyrophilic nucleolar organizer regions (AgNORs) were quantified in oral carcinomas (n = 39) with or without human papillomavirus (HPV) infection. The AgNOR counts of the HPV-positive samples (7.15 +/- 2.13) were not significantly (P = 0.09) higher than those of the HPV-negative ones (6.16 +/- 1.89). Furthermore, the lesions infected with multiple HPV types had greater counts than those with HPV type 16/18 infection alone. Significant differences were observed between the mean counts of the poorly (10.50 +/- 0.54), moderately (7.31 +/- 1.07) and well- (5.12 +/- 0.85) differentiated carcinomas. The mean AgNOR numbers in the oral carcinomas at TNM stages III/IV were found to be significantly (P < 0.01) higher than the numbers in corresponding stage II lesions. Cytokinetics of the lesions assessed by the bromodeoxyuridine (Brdu) labelling index (LI%) showed a linear correlation (r = 0.91; P < 0.0001) with their respective mean AgNOR counts.

Cytokinetics of 27 untreated oral squamous cell carcinomas were evaluated by in vitro bromodeox- yuridine (BrdU) labelling. A mononuclear cell suspension was prepared for the labelling using collagenase and DNase treatment. The labelled cells were visualised by immunofluorescence staining with an anti-BrdU monoclonal antibody. The labelling index (LI%) was calculated by determining the percentage of BrdU labelled cells. The LIs of the carcinomas ranged from 3.8 to 19.2%. The LIs of poorly-differentiated (HG3) tumours were 2–3-fold higher compared with those of well-differentiated (HG1) and moderately-differentiated (HG2) carcinomas. Results also showed statistically significant (P < 0.0005) increases in LIs from nuclear grade 1 (NG1) to nuclear grade 3 (NG3) carcinomas. Higher LIs were observed in stage III/IV (10.82 ± 4.62; 10.36 ± 4.90) than those in stage I (6.87 ± 2.09) and II (7.14 ± 1.87) carcinomas. A significant (P < 0.0005) difference in LI values was found between the patients with positive and negative lymph nodes. Good correlation (r = 0.77) was exhibited between the LI values and mitotic counts (MC) of the specimens. These results on oral cancer cell proliferation seem to have prognostic implications.

p53 protein expression in oral squamous cell carcinomas (SCCs) was studied in relation to presence of HPV capsid antigens and tumour cell kinetics. The protein and viral antigens were determined from 64 SCCs employing immunoperoxidase and immunofluorescence techniques, respectively. Cell kinetics were assessed by in vitro bromodeoxyuridine labelling. Different levels of p53 were detected in 61% cases. Well-differentiated SCCs (88%) showed increased p53 expression compared with the less differentiated ones. In SCCs with higher cell proliferation rate (LI%, 13.25 ± 4.64) the expression was almost absent. However, it was higher in stage I than in stage II/III carcinomas and occurred in 54.5% and 75% cases of HPV antigen-positive and -negative (or very little) SCCs, respectively.

To determine the association of human papillomavirus (HPV) with oral cancer lesions containing abnormal mitoses, we analysed 54 of them by immunohistochemical techniques for the presence of HPV structural antigens and correlated the results with their histology. Sixteen out of 19 lesions with abnormal mitotic figures (MFs) and 15 of the 25 lesions with normal MFs contained the HPV antigen. However, 9 of the 11 koilocytotic lesions with abnormal MFs and 13 of the 17 lesions with normal MFs were HPV antigen-positive. The nature of mitosis was not influenced by the grades of koilocytosis. No correlation of HPV antigen positivity and severity of the oral carcinomas was observed.

Abstract 1220 Despite the success of imatinib mesylate (IM), complete molecular remission is difficult to achieve in IM-treated CML patients. After continuing treatment with IM, a population of leukemic stem/progenitor cells remains, which are insensitive to IM. Our previous studies have shown that Jak2 regulates Bcr-Abl signaling and maintains levels of Bcr-Abl in CML cell lines, fresh cells from blast crisis patients, and mouse hematopoietic cells transformed by wild-type and imatinib-resistant mutants of BCR-ABL (Samanta et al. ASH 2009 meeting, submitted). We hypothesize that Jak2 plays similar role in Bcr-Abl+ CD34+ stem/progenitor cells, and that inhibition of Jak2 will induce apoptosis by inhibiting critical signaling pathways. In our current studies we have examined Bcr-Abl CD34+ progenitor cells from CML blast crisis patients. These cells are resistant to IM but quite sensitive to cell death induction by the selective Jak2 inhibitor TG101209 at 5–10 μM concentrations for 48 h. Similar results were obtained with cells from IM-resistant accelerated phase and IM-resistant chronic phase patients. Western blotting of CD34+ cells from an IM-resistant blast crisis CML patient established that a four h treatment with TG101209 at 10 μM severely reduced levels of Bcr-Abl protein and levels of phosphotyrosine 177 within Bcr-Abl. To begin to assess whether Jak2 effects on Bcr-Abl signaling also occur in early stage Bcr-Abl induced disease, we assayed CD34+ cord blood cells transformed by Bcr-Abl in short-term culture (Chu S et al. Cancer Res 2007; 67: 7045–7053). The results showed that a six h treatment with TG101209 (10 μM) reduced levels of: Jak2 protein; activated Jak2 (lower levels of pTyr 1007/8); Tyr 177 within Bcr-Abl; and the Bcr-Abl protein itself. These results support our earlier findings that Jak2 regulates Bcr-Abl signaling in CML cell lines (Samanta AK, et al. Cancer Res 66 (13):6468-72, 7/2006; Samanta AK, et al., Oncogene 28 (14):1669-81, 4/2009; Tao W., et al., Oncogene 27(22):3194-200, 5/2008). Furthermore, these results suggest that Jak2 plays a critical role in progenitor CML cells from early stage and late stage patients with CML disease. Disclosures: Bhatia: Novartis: Consultancy.

Abstract 39 We have shown that Jak2 is activated in Bcr-Abl+ hematopoietic cells. Our findings indicate that Jak2 kinase inhibition overcomes drug-resistance in CML cells (Samanta et al., 2006). Recently, we showed that Jak2 inhibition deactivates the Lyn kinase in Bcr-Abl+ cells by inhibiting SET expression; SET inhibits the PP2A-SHP1 (Samanta et al., 2009). Because of the critical effects of Jak2 in Bcr-Abl+ cells, we explored the mechanism of Jak2 action in CML cells. Bcr-Abl is known to be phosphorylated on a number of tyrosine residues including tyrosine 177. Phosphorylation of Tyr 177 leads to binding to Grb2, activation of the SOS-Ras pathway, and activation of the PI-3 kinase through the Grb2-Gab2 pathway. Here, we show that Jak2 and not Bcr-Abl phosphorylates Tyr 177 of Bcr-Abl. Tyr 177 of Bcr-Abl (YVNV) has the Jak2 consensus target sequence (YxxV/L/I) (Argetsinger et al., 2004) as do a number of other Tyr residues in the Bcr portion of Bcr-Abl (b3a2). Inhibition of Jak2 by a specific Jak2 kinase inhibitor TG101209 (TargeGen Inc., San Diego, CA) and a new Jak2 kinase inhibitor (WP1193, synthesized by Dr. Priebe) in Bcr-Abl+ cells, but not inhibition of Bcr-Abl by imatinib mesylate (IM), rapidly reduced phosphorylation of Tyr 177 of Bcr-Abl in IM-sensitive and –resistant cell lines including K562, K562-R, Bcr-Abl+ 32D, Bcr-Abl+ BaF3, Bcr-Abl+T315I BaF3 cells and cells from blast crisis CML patients. Jak2 inhibitors but not IM also strongly inhibited the phosphorylation of a synthetic Bcr peptide containing Tyr 177 (and the surrounding Bcr sequences) catalyzed by either recombinant purified Jak2 or Jak2 immune complexes. Jak2 inhibition of Bcr-Abl+ 32D cells rapidly reduced Grb2 binding to Bcr-Abl, and diminished activation of Ras and PI-3 kinase pathways within two h. Importantly, knockdown of Jak2 with Jak2-specific siRNA also reduced levels of pTyr 177 Bcr-Abl and total Bcr-Abl protein. Rescue experiments reversed Jak2 knockdown and stimulated pTyr 177 levels. In whole cells the total levels of the Bcr-Abl protein also decreased upon Jak2 inhibition with the above chemical inhibitors but at a rate slower than the rate observed in the inhibition of the Tyr 177 phosphorylation of Bcr-Abl. In kinase assays with Bcr-Abl network complexes from Bcr-Abl+ cells, phosphorylation of Tyr 177 of Bcr-Abl was drastically reduced by Jak2 inhibition but Bcr-Abl protein levels remained stable. Reduction of Bcr-Abl protein levels in Bcr-Abl+ cells by Jak2 inhibition also caused a severe reduction of tyrosine phosphorylation of STAT5 and STAT3. We have shown by gel filtration column chromatography of lysates isolated from Bcr-Abl+ cells that Bcr-Abl is physically associated with the IL-3 receptor beta chain, Jak2, Akt and HSP90 in high molecular weight complexes. GSK3β, STAT5 and STAT3 also emerge from the column in the high molecular weight fractions and suggested that several high molecular weight complexes may exist. Nevertheless, incubation of Bcr-Abl+ cells with WP1193 caused disruption of these large molecular network complexes within three h. Nude mice, injected subcutaneously with IM-resistant K562-R cells developed solid tumors, which were reduced in weight by i.p. injection of WP1193 every 48 h at 30 mg/Kg over 10 days. WP1193 at a similar dosage strongly reduced leukemic and tumor effects of Bcr-Abl+ 32D cells expressing T315I Bcr-Abl injected i.v. into nude mice. WP1193 injections into nude mice did not cause significant toxicity over a 10 day period. In summary, Jak2 inhibitors TG101209/WP1193 rapidly reduced levels of pTyr 177 Bcr-Abl, reduced levels of Bcr-Abl protein and induced high levels of apoptosis in IM-sensitive and –resistant Bcr-Abl+ cell lines including T315I cells, CML cell lines, cells from CML patients in blast crisis. In addition Jak2 inhibition reduced tumor formation in mice injected with IM-resistant K562-R and T315I Bcr-Abl+ mouse cells. These findings establish: 1) Jak2 as a major driver of Bcr-Abl signaling pathways in Bcr-Abl+ CML cells, which were previously thought to be activated directly by Bcr-Abl. 2) Although Jak2 is activated by Bcr-Abl, Jak2 is the dominant target molecule in CML and Jak2 becomes even more dominant in CML cells that become resistant to IM and other tyrosine kinase inhibitors. These findings indicate that CML cells, because of the dominant role of Jak2, resemble leukemia cells from Bcr-Abl negative myeloproliferative diseases. Disclosures: No relevant conflicts of interest to declare.

In chronic myelogenous leukemia (CML), treatment of early stage disease has been significantly improved with the discovery of imatinib mesylate (IM, Gleevec), but with the progression of the disease, drug resistance develops which is due to mutation of the ATP binding site domain within the Bcr-Abl tyrosine kinase, the activation of Lyn kinase (a Src-related tyrosine kinase) and other mutations that produce a more aggressive disease. Thus, patients relapse and the disease progresses to blast crisis (BC) stage where survival is limited to only several months. No suitable drug treatment for BC is available, and the mechanism of drug resistance for BC is unclear. A new synthesized small molecule, WP1193, a caffeic acid derivative, induced apoptosis in all types of drug-resistant CML cells including IM-resistant T315I BCR-ABL+ cells, IM-resistant CML cells with up-regulated Lyn tyrosine kinase (K562-R) and cells from blast crisis CML patients. Treatment of BCR-ABL+ cells with WP1193 inhibited Janus kinase activities, leading to strong reduction of Bcr-Abl; this treatment also reduced STAT3 and pTyr 705 STAT3 protein levels. CML cell lines contain a large signaling network complex composed of Bcr-Abl, Jak2, HSP90 and its client proteins. HSP90 is a molecular chaperone that plays an important role in conformational maturation and stabilization of client proteins (Bcr- Abl, Jak2, STAT3, etc.), which are physically associated in the signaling network complex. This network complex (estimated to be between 2–5 million daltons) has been characterized by gel filtration column chromatography. This complex is disassembled by treatment of Bcr-Abl+ cells with WP1193 (10 μM for 3–6 h). Treatment of the cells with WP1193 also inhibited binding of STAT3 to its consensus DNA sequence and reduced levels of HSP90α transcripts, HSP90α protein and its client proteins, which include Bcr-Abl, Jak2 and Akt. WP1193 also reduced solid tumors in mice inoculated with IM-resistant K562-R cells, and inhibited leukemic effects caused by T315I BCR-ABL+ cells. These results indicate that disruption of the Bcr-Abl/Jak2/HSP90 network complex by WP1193 overcomes drug resistance in advanced stage CML cells. Since disruption of this network complex causes apoptosis induction in all IM-resistant and late stage CML patient cells tested so far, and since no significant toxicity was observed in short-term mouse experiments, WP1193 may have medical utility for the treatment of IM-resistant forms of CML and late stage CML where other drugs fail. One novel feature of WP1193 is its ability to reduce the activity of several important components of the network, namely Bcr-Abl, Jak2 and STAT3 activities and reduction of HSP90α, which then causes a high level of apoptosis in IM-resistant cells and late-stage CML cells.

Background and Aims: Dysarthria affects linguistic domains such as respiration, phonation, articulation, resonance and prosody due to upper motor neuron, lower motor neuron, cerebellar or extrapyramidal tract lesions. Although Bengali is one of the major languages globally, dysarthric Bengali speech has not been subjected to neurolinguistic analysis. We attempted such an analysis with the goal of identifying the speech defects in native Bengali speakers in various types of dysarthria encountered in neurological disorders. Settings and Design: A cross-sectional observational study was conducted with 66 dysarthric subjects, predominantly middle-aged males, attending the Neuromedicine OPD of a tertiary care teaching hospital in Kolkata. Materials and Methods: After neurological examination, an instrument comprising commonly used Bengali words and a text block covering all Bengali vowels and consonants were used to carry out perceptual analysis of dysarthric speech. From recorded speech, 24 parameters pertaining to five linguistic domains were assessed. The Kruskal-Wallis analysis of variance, Chi-square test and Fisher′s exact test were used for analysis. Results: The dysarthria types were spastic (15 subjects), flaccid (10), mixed (12), hypokinetic (12), hyperkinetic (9) and ataxic (8). Of the 24 parameters assessed, 15 were found to occur in one or more types with a prevalence of at least 25%. Imprecise consonant was the most frequently occurring defect in most dysarthrias. The spectrum of defects in each type was identified. Some parameters were capable of distinguishing between types. Conclusions: This perceptual analysis has defined linguistic defects likely to be encountered in dysarthric Bengali speech in neurological disorders. The speech distortion can be described and distinguished by a limited number of parameters. This may be of importance to the speech therapist and neurologist in planning rehabilitation and further management.

A 64 year old patient was admitted for elective coronary angiography for chronic stable angina. We proceeded through the right radial approach. After cannulating the radial artery, a 5 French sheath was inserted using a 0.038 inch guidewire and TIG 5 French (Terumo) catheter. While advancing the guidewire, it was observed …

Pathogenicity of Fusarium solani f.sp. pisi (F. R. Jones) Snyd. & Hans was tested on five cultivars of pea (Pisum sativum L.) under identical conditions; the fungus was more virulent on cv. Arkel and less virulent on cv. Sweet Stringless. Bacterization of seeds with Rhizobium leguminosarum biovar vicieae was highly effective in reducing the severity of root rot of pea. In vitro tests with F. solani f.sp. pisi and R. leguminosarum biovar vicieae showed no antagonistic effects on solid medium. However, growth of both microorganisms was retarded in dual culture. Phytoalexins (pisatin and 4-hydroxy-2,3,9-trimethoxypterocarpan) were isolated from both Fusarium- and Rhizobium–Fusarium-infected pea epicotyls; 4-hydroxy-2,3,9-trimethoxypterocarpan was present in a greater amount in the latter than in the former, while pisatin concentration was similar in samples from both treatments.

Rhizobium leguminosarum IARI 102 produced 2,3-dihydroxy benzoic acid, a type of phenolate siderophore, under iron-starved conditions. Hydroxamic acids were not detected. Maximum production of siderophore was found at 26 h of growth in a chemically defined medium at 28°C with shaking. Threonine was detected as the amino acid conjugate of the siderophore. Addition of Fe3+ to the culture medium increased the growth yield significantly, but depressed the production of the iron chelating compound.