DNA and protein binding, double-strand DNA cleavage and cytotoxicity of mixed ligand copper(II) complexes of the antibacterial drug nalidixic acid.

The water soluble mixed ligand complexes [Cu(nal)(diimine)(H2O)](ClO4) 1-4, where H(nal) is nalidixic acid and diimine is 2,2'-bipyridine (1), 1,10-phenanthroline (2), 5,6-dimethyl-1,10-phenanthroline (3), and 3,4,7,8-tetramethyl-1,10-phenanthroline (4), have been isolated. The coordination geometry around Cu(II) in 1 and that in the Density Functional Theory optimized structures of 1-4 has been assessed as square pyramidal. The trend in DNA binding constants (Kb) determined using absorption spectral titration (Kb: 1, 0.79±0.1<2, 1.06±0.1<3, 1.79±0.2<4, 1.84±0.2×105M-1) is in line with that (Kapp) determined by competitive ethidium bromide binding studies. The large red-shift (10nm) observed for 2 suggests that the phen co-ligand is stacked with a frayed DNA base pair. In contrast, 3 and 4 are involved in intimate hydrophobic interaction with DNA through the methyl substituents on phen ring, which is supported by viscosity and protein binding studies. DNA docking studies imply that 4 is involved preferentially in DNA major groove binding while 1-3 in minor groove binding and that all the complexes, upon removing the axially coordinated water molecule, bind in the major groove. Interestingly, 3 and 4 display prominent double-strand DNA cleavage while 1 and 2 effect only single-strand DNA cleavage in the absence of an activator. The complexes 3 and 4 show cytotoxicity higher than 1 and 2 against human breast cancer cell lines (MCF-7). The complex 4 induces apoptotic mode of cell death in cancer cells.

Cutaneous adenocarcinoma of sebaceous gland in a captive male jaguar Panthera onca: a case report.

High incidence of neoplasia in captive jaguar (Panthera onca) has been recorded but there have been no reports of cutaneous adenocarcinoma of the sebaceous gland. A high incidence of neoplasia has been detected in captive jaguars, possibly associated with longevity and husbandry practices in captivity. Neoplasm is a major cause of mortality in jaguar. Tumours of sebaceous gland are common in older domestic felids. A case of cutaneous adenocarcinoma of the sebaceous gland was diagnosed in a male captive jaguar in the Zoological Garden, Alipore, Kolkata, India and was managed successfully. The tumour was observed as a superficial, ulcerated, multilobulated intradermal mass. After preoperative haematological evaluation the tumour was excised through routine surgical procedure under chemical immobilisation. Post-operative management was uneventful. Local tumour recurrence was not noticed till one year after post-operation.

Speciation of platinum-benzoylthiourea in the gas phase using electrospray ionization mass spectrometry and density functional theory.

RATIONALE: Determining the speciation of platinum-benzoylthiourea (Pt-BTU) in the gas phase is a challenging task due to various reaction pathways and the conformational flexibility of the BTU ligand. METHODS: Electrospray ionization mass spectrometry (ESI-MS) experiments and density functional theory (DFT) based calculations were carried out to shed light on this complex reaction in the gas phase using K2 PtCl4 salt and BTU. Various Pt complexes were studied in both positive and negative ion modes of ESI-MS using a quadrupole-time-of-flight mass spectrometer. The effects of the ESI-MS experimental parameters such as capillary voltage, pH and electrolyte on the peak intensity of the Pt-BTU complex were investigated. DFT calculations employing B3LYP functional with the 6-311++G** basis set were used to characterize the geometric parameters and fragmentation patterns of various Pt complexes in the gas phase. RESULTS: In the positive ion mode, complexes with differing numbers of BTU ligands coordinated to the metal ion were observed, whereas, in the negative ion mode, no species associated with BTU or with the solvent (acetonitrile) molecules were found. It was also found that Pt forms complexes with the BTU ligand in different stoichiometric ratios. For both Pt(BTU)2 and Pt(BTU)3 complexes, the BTU ligand undergoes deprotonation followed by bi-dentate coordination. DFT calculations suggest that BTU can coordinate to Pt in both cis and trans isomeric forms, which are nearly iso-energetic with a slight preference towards the trans-isomer. The preference of trans-BTU binding is attributed to the exclusive retention of intra-molecular hydrogen bonding which is absent in the cis-form. CONCLUSIONS: Experimental and theoretical calculations have shown that the gas-phase interaction of BTU to Pt is very complex. The BTU ligand can coordinate to Pt in both mono-dentate and bi-dentate modes, the latter mode being favorable upon deprotonation of the BTU ligand. Furthermore, many close lying species with different geometric isomeric forms are found to be possible due to the presence of intra- and inter-molecular hydrogen bonding.

Effects of small intestinal submucosa (SIS) on the murine innate immune microenvironment induced by heat-killed Staphylococcus aureus.

The use of biological scaffold materials for wound healing and tissue remodeling has profoundly impacted regenerative medicine and tissue engineering. The porcine-derived small intestinal submucosa (SIS) is a licensed bioscaffold material regularly used in wound and tissue repair, often in contaminated surgical fields. Complications and failures due to infection of this biomaterial have therefore been a major concern and challenge. SIS can be colonized and infected by wound-associated bacteria, particularly Staphylococcus aureus. In order to address this concern and develop novel intervention strategies, the immune microenvironment orchestrated by the combined action of S. aureus and SIS should be critically evaluated. Since the outcome of tissue remodeling is largely controlled by the local immune microenvironment, we assessed the innate immune profile in terms of cytokine/chemokine microenvironment and inflammasome-responsive genes. BALB/c mice were injected intra-peritoneally with heat-killed S. aureus in the presence or absence of SIS. Analyses of cytokines, chemokines and microarray profiling of inflammasome-related genes were done using peritoneal lavages collected 24 hours after injection. Results showed that unlike SIS, the S. aureus-SIS interactome was characterized by a Th1-biased immune profile with increased expressions of IFN-γ, IL-12 and decreased expressions of IL-4, IL-13, IL-33 and IL-6. Such modulation of the Th1/Th2 axis can greatly facilitate graft rejections. The S. aureus-SIS exposure also augmented the expressions of pro-inflammatory cytokines like IL-1ß, Tnf-α, CD30L, Eotaxin and Fractalkine. This heightened inflammatory response caused by S. aureus contamination could enormously affect the biocompatibility of SIS. However, the mRNA expressions of many inflammasome-related genes like Nlrp3, Aim2, Card6 and Pycard were down-regulated by heat-killed S. aureus with or without SIS. In summary, our study explored the innate immune microenvironment induced by the combined exposure of SIS and S. aureus. These results have practical implications in developing strategies to contain infection and promote successful tissue repair.

Rhinoplasty in atrophic rhinitis.

Rhinoplasty in atrophic rhinitis is a difficult surgery because the dorsal skin is adherent to the underlying structures. There is also more chance of postoperative infection. The experience of various types of rhinoplasty in 15 atrophic rhinitis patients is presented here. The patients were from the age group 17 to 35 years. Most of the operations were done under local anaesthesia. Commonest graft used was conchal-cartilage. Bone graft was avoided for augmentation because of its high rate of absorption in atrophic rhinitis. Young's operation was done in 5 patients in one side along with rhinoplasty. Young's operation was done only with skin layer.

Extracellular matrix from porcine small intestinal submucosa (SIS) as immune adjuvants.

Porcine small intestinal submucosa (SIS) of Cook Biotech is licensed and widely used for tissue remodeling in humans. SIS was shown to be highly effective as an adjuvant in model studies with prostate and ovarian cancer vaccines. However, SIS adjuvanticity relative to alum, another important human-licensed adjuvant, has not yet been delineated in terms of activation of innate immunity via inflammasomes and boosting of antibody responses to soluble proteins and hapten-protein conjugates. We used ovalbumin, and a hapten-protein conjugate, phthalate-keyhole limpet hemocyanin. The evaluation of SIS was conducted in BALB/c and C57BL/6 mice using both intraperitoneal and subcutaneous routes. Inflammatory responses were studied by microarray profiling of chemokines and cytokines and by qPCR of inflammasomes-related genes. Results showed that SIS affected cytokine and chemokines microenvironments such as up-regulation of IL-4 and CD30-ligand and activation of chemotactic factors LIX and KC (neutrophil chemotactic factors), MCP-1 (monocytes chemotactic factors), MIP 1-α (macrophage chemotactic factor) and lymphotactin, as well as, growth factors like M-CSF. SIS also promoted gene expression of Nod-like receptors (NLR) and associated downstream effectors. However, in contrast to alum, SIS had no effects on pro-inflammatory cytokines (IL-6, IL-1ß, TNF-α) or NLRP3, but it appeared to promote both Th1 and Th2 responses under different conditions. Lastly, it was as effective as alum in engendering a lasting and specific antibody response, primarily of IgG1 type.

Synthetic adjuvants for vaccine formulations: evaluation of new phytol derivatives in induction and persistence of specific immune response.

Terpenoids are ubiquitous natural compounds that have been shown to improve vaccine efficacy as adjuvants. To gain an understanding of the structural features important for adjuvanticity, we studied compounds derived from a diterpene phytol and assessed their efficacy. In a previous report, we showed that phytol and one of its derivatives, PHIS-01 (a phytol-derived immunostimulant, phytanol), are excellent adjuvants. To determine the effects of varying the polar terminus of PHIS-01, we designed amine and mannose-terminated phytol derivatives (PHIS-02 and PHIS-03, respectively). We studied their relative efficacy as emulsions with soluble proteins, ovalbumin, and a hapten-protein conjugate phthalate-KLH. Immunological parameters evaluated consisted of specific antibody responses in terms of titers, specificities and isotype profiles, T cell involvement and cytokine production. Our results indicate that these new isoprenoids were safe adjuvants with the ability to significantly augment immunogen-specific IgG1 and IgG2a antibody responses. Moreover, there was no adverse phthalate cross-reactive anti-DNA response. Interestingly, PHIS-01 and PHIS-03 influenced differentially T-helper polarization. We also observed that these compounds modulated the immune response through apoptotic/necrotic effects on target tumor cells using murine lymphomas. Finally, unlike squalene and several other terpenoids reported to date, these phytol derivatives did not appear arthritogenic in murine models.

The study of the ideal curvature of metallic tracheostomy tube in ca larynx.

Twenty patients in the age group of 45-55 years with carcinoma of larynx underwent tracheostomy for acute upper airway obstruction. Tracheostomy tubes (no. 36) commonly available in the market have varying angle of curvatures ranging from 120° to 130°. In respect of different curvatures of the tracheostomy tubes, the various complaints of the patients were noted. The mean ideal angle of curvature of the tracheostomy tube should be 112° that was calculated from CT scan of neck with tube in situ.

Ab initio studies on the electronic structure and properties of aluminum hydrides that are analogues of boron hydrides.

Although the boron hydrides are well-known in the literature, the aluminum hydride chemistry is limited to very few systems such as AlH(3), its dimer, and its polymeric form. In view of the recent experimental studies on the possible existence of the aluminum hydrides, herein, we have undertaken a systematic study on the electronic structure and properties of these aluminum hydrides. Under this, we have studied different classes of hydrides, viz., closo (Al(n)H(n+2)), nido (Al(n)H(n+4)), and arachno (Al(n)H(n+6)), similar to the boranes. All the aluminum hydrides are found to have exceptionally large highest-occupied molecular orbital-lowest-unoccupied molecular orbital gaps, low electron affinities, large ionization potentials and also large enthalpy and free energy of atomization. In addition, most of the structures are also found to have high symmetries. These exceptional properties can be indicative of the pronounced stability, and hence, it is expected that other aluminum hydride complexes can indeed be observed experimentally.

A novel cell-surface protein CSP82 on bone marrow stem cells and a cytosolic phosphoprotein DP58 (ankyrinRD 34B) are involved in promyeloid progenitor induction.

The molecular events associated with the development of common myeloid progenitor (CMP) remain largely unknown. This study reports that a novel glycosylphosphatidylinositol (GPI)-anchored lactoferrin CSP82 on uninitiated mouse bone marrow cells (BMC) may be involved in inducing pro-DC from CMP. By peptide mass fingerprinting, CSP82 has been identified as the mouse lactoferrin precursor, but unlike the latter, it occurs as a GPI-linked cell-surface protein. The GPI-linkage was demonstrated on BMC-derived immunoprecipitates and by other techniques. Furthermore, BMC and hematopoietic stem BM cells following incubation with either CSP82 peptide antibody or purified Reagent A yielded CMP-like progenitors (BM4 cells). These progenitors expressed a previously reported cytosolic phosphoprotein DP58 (AnkRD 34B protein). Continued cultivation of BMC in media containing only anti-CSP82 antibody led to DC-like cells, that bore phenotypic and endocytic resemblance with those obtained using GM-CSF. The results suggest that a receptor lactoferrin on BMC may be an important non-cytokine mechanism for early promyeloid progenitor differentiation.