Oxidative inhibition of Hsp90 disrupts the super‐chaperone complex and attenuates pancreatic adenocarcinoma in vitro and in vivo

Pancreatic cancer is almost always fatal, in part because of its delayed diagnosis, poor prognosis, rapid progression and chemoresistance. Oncogenic proteins are stabilized by the Hsp90, making it a potential therapeutic target. We investigated the oxidative stress‐mediated dysfunction of Hsp90 and the hindrance of its chaperonic activity by a carbazole alkaloid, mahanine, as a strategic therapeutic in pancreatic cancer. Mahanine exhibited antiproliferative activity against several pancreatic cancer cell lines through apoptosis. It induced early accumulation of reactive oxygen species (ROS) leading to thiol oxidation, aggregation and dysfunction of Hsp90 in MIAPaCa‐2. N‐acetyl‐L ‐cysteine prevented mahanine‐induced ROS accumulation, aggregation of Hsp90, degradation of client proteins and cell death. Mahanine disrupted Hsp90‐Cdc37 complex in MIAPaCa‐2 as a consequence of ROS generation. Client proteins were restored by MG132, suggesting a possible role of ubiquitinylated protein degradation pathway. Surface plasmon resonance study demonstrated that the rate of interaction of mahanine with recombinant Hsp90 is in the range of seconds. Molecular dynamics simulation showed its weak interactions with Hsp90. However, no disruption of the Hsp90‐Cdc37 complex was observed at an early time point, thus ruling out that mahanine directly disrupts the complex. It did not impede the ATP binding pocket of Hsp90. Mahanine also reduced in vitro migration and tube formation in cancer cells. Further, it inhibited orthotopic pancreatic tumor growth in nude mice. Taken together, these results provide evidence for mahanine‐induced ROS‐mediated destabilization of Hsp90 chaperone activity resulting in Hsp90‐Cdc37 disruption leading to apoptosis, suggesting its potential as a specific target in pancreatic cancer.     

Outcomes of phacoemulsification in patients with dry eye

PURPOSE: To evaluate the outcomes of phacoemulsification in patients with dry eye. SETTING: Department of Ophthalmology, Post Graduate Institute of Medical Education and Research, Chandigarh, India. METHODS: This study included 25 eyes of 23 patients with dry eye having phacoemulsification. Dry eye was defined as Schirmer I with lidocaine hydrochloride (Xylocaine) score of 5.0 mm after 5 minutes, a tear-film breakup time (TFBUT) of less than 5 seconds, or both. Data were retrospectively analyzed for preoperative and postoperative tear function, postoperative complications, and final visual outcomes. RESULTS: Of the 23 patients, 18 had age-related dry eye and 5 had secondary Sj?gren's syndrome. Twenty-two eyes had predominant aqueous deficiency (Schirmer I with Xylocaine score of 5.0 mm or less), and 3 had a Schirmer score between 6.0 mm and 9.0 mm. The TFBUT was 5 seconds or less in 17 eyes and between 6 seconds and 9 seconds in 8 eyes. The mean preoperative Schirmer score was 4.80 mm +/- 2.01 (SD) (range 2.0 to 9.0 mm) and the mean postoperative score, 3.80 +/- 2.40 mm (range 0 to 7.0 mm). The mean preoperative TFBUT was 4.00 +/- 1.87 seconds (range 0 to 9 seconds) and the mean score at the last follow-up, 3.40 +/- 1.60 seconds (range 0 to 8 seconds). Postoperatively, 8 eyes had superficial punctate keratopathy and 8 had an epithelial defect. The final visual acuity was 6/6 in 13 eyes, 6/9 to 6/12 in 8 eyes, and 6/18 to 6/60 in 4 eyes. CONCLUSION: Phacoemulsification was safe and led to minimal complications in patients with age-related dry eye with or without associated systemic disorders.     

In vitro antimalarial activity of medicinal plant extracts against Plasmodium falciparum

Malaria is a major global public health problem, and the alarming spread of drug resistance and limited number of effective drugs now available underline how important it is to discover new antimalarial compounds. In the present study, ten plants were extracted with ethyl acetate and methanol and tested for their antimalarial activity against chloroquine (CQ)-sensitive (3D7) and CQ-resistant (Dd2 and INDO) strains of Plasmodium falciparum in culture using the fluorescence-based SYBR Green assay. Plant extracts showed moderate to good antiparasitic effects. Promising antiplasmodial activity was found in the extracts from two plants, Phyllanthus emblica leaf 50% inhibitory concentration (IC₅₀) 3D7: 7.25 μg/mL (ethyl acetate extract), 3.125 μg/mL (methanol extract), and Syzygium aromaticum flower bud, IC₅₀ 3D7:13 μg/mL, (ethyl acetate extract) and 6.25 μg/mL (methanol extract). Moderate activity (30–75 μg/mL) was found in the ethyl acetate and methanol extracts of Abrus precatorius (seed) and Gloriosa superba (leaf); leaf ethyl acetate extracts of Annona squamosa and flower of Musa paradisiaca. The above mentioned plant extracts were also found to be active against CQ-resistant strains (Dd2 and INDO). Cytotoxicity study with P. emblica leaf and S. aromaticum flower bud, extracts showed good therapeutic indices. These results demonstrate that leaf ethyl acetate and methanol extracts of P. emblica and flower bud extract of S. aromaticum may serve as antimalarial agents even in their crude form. The isolation of compounds from P. emblica and S. aromaticum seems to be of special interest for further antimalarial studies.     

Development of mechanical hypersensitivity in rats during heroin and ethanol dependence: alleviation by CRF₁ receptor antagonism

Animal models of drug dependence have described both reductions in brain reward processes and potentiation of stress-like (or anti-reward) mechanisms, including a recruitment of corticotropin-releasing factor (CRF) signaling. Accordingly, chronic exposure to opiates often leads to the development of mechanical hypersensitivity. We measured paw withdrawal thresholds (PWTs) in male Wistar rats allowed limited (short access group: ShA) or extended (long access group: LgA) access to heroin or cocaine self-administration, or in rats made dependent on ethanol via ethanol vapor exposure (ethanol-dependent group). In heroin self-administering animals, after transition to LgA conditions, thresholds were reduced to around 50% of levels observed at baseline, and were also significantly lower than thresholds measured in animals remaining on the ShA schedule. In contrast, thresholds in animals self-administering cocaine under either ShA (1 h) or LgA (6 h) conditions were unaltered. Similar to heroin LgA rats, ethanol-dependent rats also developed mechanical hypersensitivity after eight weeks of ethanol vapor exposure compared to non-dependent animals. Systemic administration of the CRF1R antagonist MPZP significantly alleviated the hypersensitivity observed in rats dependent on heroin or ethanol. The emergence of mechanical hypersensitivity with heroin and ethanol dependence may thus represent one critical drug-associated negative emotional state driving dependence on these substances. These results also suggest a recruitment of CRF-regulated nociceptive pathways associated with escalation of intake and dependence. A greater understanding of relationships between chronic drug exposure and pain-related states may provide insight into mechanisms underlying the transition to drug addiction, as well as reveal new treatment opportunities. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.     

Multifunctional silica nanotubes for dual-modality gene delivery and MR imaging

This work demonstrated the development of multifunctional silica nanotubes (SNT) by functionalization of their inner void and outer surface with magnetic-fluorescence nanocomposites and cationic polymers, respectively. The successful construction of BPEI-SNT was established by electron energy loss spectroscopy in conjugation with standard analytical tools. The mean fluorescence intensity in a FACS assay, a luciferase gene expression assay and a confocal fluorescence study demonstrated the efficacy of BPEI-SNT as a gene delivery vector. Endocytotic uptake was also demonstrated by the colocalization of LysoTracker Red(?) and green fluorescent quantum dots. Moreover, enhanced magnetic resonance imaging revealed the potential of the BPEI-SNT nanocomposite to act as a dual-modality nano-device.     

Pandemic and seasonal influenza viruses among patients with acute respiratory illness in Kashmir (India)

Please cite this paper as: Koul PA., et al. (2011) Pandemic and seasonal influenza viruses among patients with acute respiratory illness in Kashmir (India). Influenza and Other Respiratory Viruses 5(6), e521–e527. Background With the emergence of pandemic influenza A (2009A/H1N1) virus in India, we sought to determine the prevalence and clinical presentations of seasonal and pandemic influenza viruses among acute respiratory illness (ARI) patients from Srinagar, a temperate climate area in northern India, during the peak winter season. Methods Combined throat and nasal swabs, obtained from 194 (108 male) presenting with ARI from January to March 2010 (Week 53‐week 10), were tested by RT‐PCR for influenza A and B, including 2009A/H1N1 viruses. HA1 gene of selected 2009A/H1N1‐positive samples was sequenced, and phylogenetic analysis was carried out. Results Twenty‐one (10·8%, age 15–80 years, median age 40 years) patients tested positive for influenza viruses: 13 (62%) for 2009A/H1N1 virus, 6 (28·5%) for seasonal influenza A (H3N2), and 2 (9·5%) for influenza B. Twelve of the 13 patients with 2009A/H1N1 presented with febrile ARI, and eight had associated comorbidities. All of the patients recovered. Phylogenetic analysis of HA gene (n = 8) revealed that all strains from Srinagar clustered in 2009A/H1N1 clade seven along with the other 2009A/H1N1 strains from India. Amino acid substitutions in the HA protein defining clade seven (P83S, S203T, and I321V) were found in almost all isolates from Srinagar. Conclusions Both seasonal and 2009A/H1N1 viruses appear to be associated with ARI in Srinagar. The 2009A/H1N1 in Srinagar is genetically similar to globally circulating clade 7 strains, with unique signature sequences in the HA gene. Further investigations into ascertain the role of these mutations in possible alteration of the virulence and transmissibility of the virus are needed.     

Emergence of 2009A/H1N1 cases in a tertiary care hospital in New Delhi, India

Please cite this paper as: Broor et al. (2011) Emergence of 2009A/H1N1 cases in a tertiary care hospital in New Delhi, India. Influenza and Other Respiratory Viruses 5(6), e552–e557. Objective To determine virologic and epidemiologic characteristics of pandemic (H1N1) 2009 at All India Institute of Medical Sciences (AIIMS) a tertiary care hospital in New Delhi, India. Methods Nasal and throat swabs from patients with febrile acute respiratory illness (FARI) from August to December 2009 (n = 1401) were tested for 2009A/H1N1 and seasonal influenza A viruses by real‐time RT‐PCR. Results Of 1401 samples tested, 475 (33·9%) were positive for influenza A, of these majority (412; 87%) were 2009A/H1N1, whereas the remaining 63 (13%) were seasonal influenza A (49 were A/H3 and 14 were A/H1). While co‐circulation of 2009A/H1N1 and A/H3 was observed in August–September, subsequent months had exclusive pandemic influenza activity (October–December 2009). Pandemic 2009A/H1N1 emergence did not follow typical seasonal influenza seasonality in New Delhi, which normally peaks in July–August, but instead showed bimodal peaks in weeks 39 and 48 in 2009. The percent of specimens testing positive for 2009A/H1N1 influenza virus was found to be highest in >5‐ to 18‐year age group (41·2%; OR = 2·3; CI = 1·6–3·2; P = 0·00). Conclusions Taken together, our data provide high prevalence of pandemic 2009A/H1N1 in urban New Delhi with bimodal peaks in weeks 39 and 48 and highest risk group being the children of school‐going age (aged >5–18).