Immunobiology of Toll-like receptors: emerging trends

Toll-like receptors (TLR), a family of evolutionarily conserved pathogen recognition receptors, play pivotal role as primary sensors of invading pathogens. TLR identify molecular motifs of infectious agents (pathogen associated molecular patterns) and elicit an effective defensive response against them. Mammalian TLR derive their name from the Drosophila Toll protein, with which they share sequence similarity. So far, expression of 10 TLR is known in humans. The adaptor proteins, myeloid differentiation factor 88 and Toll IL-1 receptor (TIR) domain containing adaptor inducing IFN-beta (TRIF) are the key players in the TLR signalling cascade leading to the activation of nuclear factor (NF)-kappaB and interferon regulatory factor-3, respectively. Targeted manipulation of the TLR signalling pathway has immense therapeutic potential and may eventually prove to be a boon in the development of innovative treatments for diverse disease conditions. There is accumulating evidence that TLR agonists have tremendous potential as novel therapeutic targets. In this review, we have discussed the immunobiology of TLR and emphasize significant advances made within the ever-expanding field of TLR that provide intriguing insights efficacious in unravelling the complexities associated with TLR.     

Bio-ethanol from water hyacinth biomass: An evaluation of enzymatic saccharification strategy

Biomass feedstock having less competition with food crops are desirable for bio-ethanol production and such resources may not be localized geographically. A distributed production strategy is therefore more suitable for feedstock like water hyacinth with a decentralized availability. In this study, we have demonstrated the suitability of this feedstock for production of fermentable sugars using cellulases produced on site. Testing of acid and alkali pretreatment methods indicated that alkali pretreatment was more efficient in making the sample susceptible to enzyme hydrolysis. Cellulase and β-glucosidase loading and the effect of surfactants were studied and optimized to improve saccharification. Redesigning of enzyme blends resulted in an improvement of saccharification from 57% to 71%. A crude trial on fermentation of the enzymatic hydrolysate using the common baker’s yeast Saccharomyces cerevisiae yielded an ethanol concentration of 4.4 g/L.     

A multiply charged tetracaine derivative blocks cyclic nucleotide-gated channels at subnanomolar concentrations

Cyclic nucleotide-gated (CNG) ion channels are central participants in sensory transduction, generating the electrical response to light in retinal photoreceptors and to odorants in olfactory receptors. They are expressed in many other tissues where their specific roles in signaling remain unclear. As is true for many other ion channels, there is a paucity of specific blockers needed to dissect the contributions of these channels to cell signaling. CNG channels are members of the superfamily of voltage-gated ion channels, and the local anesthetic tetracaine is known to block CNG channels in a manner that resembles the block of voltage-gated Na(+) channels. The amine in local anesthetics interacts with the charged selectivity filter of Na(+) channels, while the aromatic ring gets stuck in the inner cavity and has hydrophobic interactions with the residues lining that region. Here we have synthesized a derivative of tetracaine, 3-[(aminopropyl)amino]-N,N-dimethyl-N-(2-[[4-(butylamino)benzoyl]oxy]ethyl)propan-1-aminium acetate (APPA-tetracaine), that contains three positively charged amines at physiological pH instead of one. This compound blocked several different CNG channels in the picomolar to nanomolar concentration range at positive membrane potentials, making it several orders of magnitude more potent than tetracaine. In contrast, significant block of Na(+) channels by APPA-tetracaine required concentrations of hundreds of nanomolar. The results suggest that the highly charged moiety of APPA-tetracaine interacts strongly with the negative charge cluster in the selectivity filter of CNG channels. We propose that a variety of potent and specific ion channel blockers could be generated by expanding on traditional blocker structures to target the selectivity filters of other channels.     

Reversal of cerulenin-induced inhibition of phospholipids and sterol synthesis by exogenous fatty acids/sterols in Epidermophyton floccosum

Cerulenin, a specific inhibitor of fatty acids and sterol biosynthesis inhibited the growth of Epidermophyton floccosum, which was reversed when growth medium was supplemented with palmitic acid and sterols. Unsaturated fatty acids partially restored the growth. Cerulenin inhibited both phospholipid and sterol biosynthesis (60-70%) at the minimum inhibitory concentration (0.5 microgram/ml) as demonstrated by [32P]orthophosphoric acid and [14C]acetate incorporation into the respective lipids. Cerulenin-induced inhibition of phospholipid and sterol synthesis was dose dependent up to 0.5 microgram/ml. Exogenously supplied fatty acids and sterols restored the biosynthesis of phospholipids in cerulenin-treated cultures, while that of sterols was enhanced. The biosynthesis of both saturated and unsaturated fatty acids was inhibited by cerulenin.     

Fatty acid composition of the postmortem prefrontal cortex of adolescent male and female suicide victims

Prior epidemiological, prospective intervention, and peripheral and central fatty acid composition studies suggest that omega-3 fatty acid deficiency may be associated with the pathoaetiology of depression and suicide. In the present study, we determined the fatty acid composition of the postmortem prefrontal cortex (PFC) of adolescent male and female suicide victims and age-matched controls. Fatty acid composition (wt% total fatty acids) and concentrations (μmol/g) were determined in the postmortem PFC (Brodmann area 10) of male and female adolescent (aged 13–20 years) suicide victims (n=20) and age-matched controls (n=20) by gas chromatography. None of the major polyunsaturated fatty acids including the principle brain omega-3 fatty acid, docosahexaenoic acid (DHA), monounsaturated fatty acids, or saturated fatty acids differed significantly between adolescent suicide victims and controls before or after segregation by gender. The arachidonic acid (AA, 20:4n-6): DHA ratio and adrenic acid (22:4n-6) composition were negatively correlated with age at death in controls but not in suicides, and males exhibited a greater AA:DHA ratio irrespective of cause-of-death. These results demonstrate that adolescent male and female suicide victims do not exhibit DHA deficits in the postmortem PFC relative to age-matched controls, and suggest that suicide victims do not exhibit the normal age-related decrease in adrenic acid composition and the AA:DHA ratio.     

Photoperiodic regulation of seasonal responses in Indian weaver bird (Ploceus philippinus)

Photoperiod (=day length) is the vital factor for the regulation of behavioral and physiological activities in many avian species. This study investigated the seasonal cycles of testicular growth and secondary sexual characteristics of Indian weaver bird under natural day length (NDL) and the effects of duration and intensity of light on photoperiodic induction. In the first experiment, groups of birds (n = 7 each) were exposed to under NDL in April 2008 and May 2009 for 8 and 12 months, respectively. In second and third experiment, birds (n = 6 each group) were exposed to different photoperiods (11.5L:12.5D, 12L:12D, 13L:11D, and 15L:9D) at the same (500 lux) light intensity, and to 13L:11D at different light intensities (10-, 50-, 500-, and 800-lux). Observations on testis size, molt, and plumage score were recorded 2-week (molt and plumage) or at 4-week intervals (testes). Both the NDL groups showed similar seasonal cycles of testicular growth-regression and secondary sexual characteristics. Second and third experiments suggest that the photoperiodic induction was depending upon duration and intensity of the light. Birds showed testicular growth-regression cycle followed by molt and plumage color change only under 13L:11D and 15L:9D and only 500- and 800-lux under 13L:11D photoperiod but not under 11.5L:12.5D and 12L:12D and 10- and 50-lux light intensities. Pre- and post-nuptial molting on body feathers were progressed with gonadal stimulation–maturation and regression cycle under 13L:11D and 15L:9D. Results under different light–dark cycles suggest that day length of about 12 h or more and above the threshold level of light intensity are essential for the induction of photoperiodic responses.     

Anatomical basis for the clinical application of the arterial supply of musculus pectoralis major.

The pattern of arterial supply to the various parts (clavicular, sternocostal and aponeurotic) of the pectoralis major muscle was studied in 7 cadaver dissections and 10 angiograms by injecting a radio-opaque substance. Three main arteries supplied the muscle, i.e. the pectoral branch of the thoracoacromial trunk (TAT-PB), the lateral thoracic artery and the perorating branch of the internal thoracic artery, supported by other branches of the TAT and the superior thoracic artery. It is observed that the TAT-PB, a chief vascular pedicle, anastomoses freely with other arteries and supplies most parts of the muscle. The present study is mainly focussed on the exclusion of the chief vascular pedicle of muscle to eliminate the confusion of previous studies and prevent the unnecessary hindrance and complications of the muscle flap.     

Ethanol Biosensors and Electrochemical Oxidation of NADH

Comparative studies of the electrochemical oxidation of reduced nicotinamide coenzyme (NADH) at the surfaces of chemically modified graphite paste electrodes (CMEs) are reported. Three different electroactive materials, tetracyanoquinodimethane (TCNQ), tetrathiafulvalene (TTF), and dimethyl ferrocene (dmFc), were used to construct three different chemically modified paste electrodes. The oxidation of NADH was examined on the basis of cyclic voltammetric measurements. The results show that all three mediators (TCNQ, TTF, and dmFc) behave as efficient mediators of the oxidation of NADH. The typical response curves of NADH at the CMEs surfaces are reported. Incorporating alcohol dehydrogenase and electroactive materials (TCNQ, TTF, and dmFc) within the graphite paste electrodes has led to the development of ethanol biosensors. Typical response curves for the ethanol analysis are reported. Comparative studies on the mediated electrochemical responses of the biosensors to ethanol are discussed.     

Interaction of hematopoietic progenitor kinase 1 and c-Abl tyrosine kinase in response to genotoxic stress

The c-Abl protein tyrosine kinase is activated by certain DNA-damaging agents and regulates induction of the stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). The hematopoietic progenitor kinase 1 (HPK1) has also been shown to act upstream to the SAPK/JNK signaling pathway. We report here that exposure of hematopoietic Jurkat T cells to genotoxic agents is associated with activation of HPK1. The results demonstrate that exposure of Jurkat cells to DNA-damaging agents is associated with translocation of active c-Abl from nuclei to cytoplasm and binding of c-Abl to HPK1. Our findings also demonstrate that c-Abl phosphorylates HPK1 in cytoplasm and stimulates HPK1 activity. The functional significance of the c-Abl-HPK1 interaction is supported by the demonstration that this complex regulates SAPK/JNK activation. Overexpression of c-Abl(K-R) inhibits HPK1-induced activation of SAPK/JNK. Conversely, the dominant negative mutant of HPK1 blocks c-Abl-mediated induction of SAPK/JNK. These findings indicate that activation of HPK1 and formation of HPK1/c-Abl complexes are functionally important in the stress response of hematopoietic cells to genotoxic agents.