Sesterterpene glycinyl-lactams: a new class of glycine receptor modulator from Australian marine sponges of the genus Psammocinia.

Bioassay guided fractionation of three southern Australian marine sponges of the genus Psammocinia, selected for their ability to modulate glycine-gated chloride channel receptors (GlyRs), yielded the rare marine sesterterpenes (-)-ircinianin (1) and (-)-ircinianin sulfate (2), along with the new biosynthetically related metabolites (-)-ircinianin lactam A (3), (-)-ircinianin lactam A sulfate (4), (-)-oxoircinianin (5), (-)-oxoircinianin lactam A (6) and (-)-ircinianin lactone A (7). Acetylation of 1 returned (-)-ircinianin acetate (8). Whole cell patch-clamp electrophysiology on 1-8 established 3 as an exceptionally potent and selective α3 GlyR potentiator, and 6 as a selective α1 GlyR potentiator. The discovery and characterization of sesterterpenes 1-8, and in particular the glycinyl-lactams 3 and 6, provide valuable new insights into GlyR pharmacology. These insights have the potential to inform and inspire the development of new molecular tools to probe GlyR distribution and function, and therapeutics to treat a wide array of GlyR mediated diseases and disorders.

Australian marine sponge alkaloids as a new class of glycine-gated chloride channel receptor modulator.

Chemical analysis of a specimen of the sponge Ianthella cf. flabelliformis returned two new sesquiterpene glycinyl lactams, ianthellalactams A (1) and B (2), the known sponge sesquiterpene dictyodendrillin (3) and its ethanolysis artifact ethyl dictyodendrillin (4), and five known sponge indole alkaloids, aplysinopsin (5), 8E-3'-deimino-3'-oxoaplysinopsin (6), 8Z-3'-deimino-3'-oxoaplysinopsin (7), dihydroaplysinopsin (8) and tubastrindole B (9). The equilibrated mixture 6/7 exhibited glycine-gated chloride channel receptor (GlyR) antagonist activity with a bias towards α3 over α1 GlyR, while tubastrindole B (9) exhibited a bias towards α1 over α3 GlyR. At low- to sub-micromolar concentrations, 9 was also a selective potentiator of α1 GlyR, with no effect on α3 GlyR-a pharmacology that could prove useful in the treatment of movement disorders such as spasticity and hyperekplexia. Our investigations into the GlyR modulatory properties of 1-9 were further supported by the synthesis of a number of structurally related indole alkaloids.

Enhancing bovine tuberculosis screening at Dhaka city in Bangladesh: Integrating gamma interferon blood test as ancillary testing with tuberculin skin test.

Tuberculin skin test (TST) is the standard method for screening of bovine tuberculosis (bTB). However, gamma interferon blood test has been introduced in the bTB control program as an ancillary testing with TST in many countries of the world. The objective of this study was to recommend this screening test as an ancillary testing with TST for field application in Bangladesh. In this study 577 cattle of different age, sex and breeds from twenty nine (29) cattle herds were examined to determine skin response against bTB through single intradermal comparative tuberculin test (SICTT) that comprised of positive (n = 81), inconclusive (n = 44) and negative (n = 452) animals. Of which 74 animals that included positive (n = 63), inconclusive (n = 8) and negative (n = 3) animals were taken under this study. Blood samples were collected in heparinized tube and stimulated overnight with bovine and avian purified protein derivatives (PPDs) for the secretion of gamma interferon, and measured via sandwich ELISA. Cohen's kappa statistics was performed for the evaluation of agreement between the two tests. The agreement obtained between two tests was fair (Kappa agreement, K = 24.0%, 95% CI = 16.9-30.5%, P = 0.037). Of positive (n = 63), inconclusive (n = 8) and negative (n = 3) status of animals at SICTT, 82.54% (n = 52), 62.50% (n = 5), and 33.33% (n = 1) were found to be bTB positive respectively through this ancillary test. This test notably corroborates to TST result. A considerable number of inconclusive TB status animals were found to be positive through this gamma interferon assay. Therefore, this test could be used as an ancillary test with TST to maximize the proportion of bTB estimation in the infected cattle herd for early detection of zoonotic tuberculosis in Bangladesh before transmission at the animal-human interface.

Gamma-interferon assay for the ancillary diagnosis of bovine tuberculosis in dairy cattle in urban and adjacent areas of Dhaka city, Bangladesh.

Background and Aim: Bovine tuberculosis (bTB) is an infectious disease of cattle, mainly caused by Mycobacterium bovis. This study aimed to evaluate the efficacy of the interferon-gamma (IFN-γ) assay and single-intradermal comparative tuberculin test (SICTT) in detecting bTB. Materials and Methods: In an earlier study, 150 positive, 83 inconclusive, and 480 negative animals from 24 cattle herds were screened using SICTT. From these groups, 125 positive, 17 inconclusive, and six negative animals were subsequently verified using the IFN-γ assay. Single-intradermal comparative tuberculin test outcomes were interpreted according to standard guidelines, whereas blood samples were collected and stimulated with purified protein derivatives. Sandwich enzyme-linked immunosorbent assay was used to measure secreted IFN-γ. Concordant and Bayesian latent class analyses were performed to evaluate test performance. Results: Results from the IFN-γ assay revealed that 83.2%, 64.7%, and 16.67% of the animals were positive in the SICTT-positive, inconclusive, and negative animal categories, respectively. Sensitivity (SE) and specificity (SP) of SICTT were 83.9% (95% confidence interval [CI]: 77.4-90.1) and 95.7% (95% CI: 86.9-99.7), respectively. Sensitivity and SP for the IFN-γ assay were 78.9% (95% CI: 71.9-85.4) and 83.9% (65.9-95.9), respectively. The use of both tests in parallel increases the SE of bTB detection (~94%), compared with SICTT alone. Conclusion: Use of the IFN-γ assay with SICTT in parallel, predominantly on cattle demonstrating an inconclusive SICTT outcome, boosts bTB detection rate in low resource settings.

Influence of Sn doping on the optoelectronic properties of ZnO nanoparticles.

Zinc Oxide (ZnO) nanoparticles (NPs) obtained a lot of attention from researchers and industries because of their superior properties as an optoelectronic material. Doping, especially tin (Sn), can further fine-tune their optoelectronic properties. In this manuscript, we have reported the optoelectronic properties of Sn-doped ZnO NPs, which were synthesized by a simple chemical solution method. A wide range of dopant (Sn) concentrations were used in the ratios of 0, 1, 3, 5, 7, and 10 weight percent. The effects of dopant (Sn) concentration on the structural, morphological, elemental composition, and optical properties of ZnO NPs were investigated by using an X-ray diffractometer (XRD), Field Emission Scanning Electron Microscope (FESEM), X-ray photoelectron spectrometer (XPS) and UV-Vis-NIR respectively. XRD analysis revealed the shifting of diffraction patterns towards a higher angle along with decreasing intensity. The calculated crystallite size using the XRD varied from 40.12 nm to 28.15 nm with an increasing doping percentage. Sn doping notably influences the size of ZnO NPs, along with crystal quality, strain, and dislocation density. The X-ray photoelectron spectroscopy (XPS) study showed the presence of zinc (Zn), oxygen (O), and tin (Sn) with their preferred oxidation states in the synthesized NPs. UV-Visible spectroscopy (UV-Vis) showed that the bandgap changed from 3.55 to 3.85 eV with the increasing concentration of Sn. FE-SEM revealed that the structures and surfaces were irregular and not homogeneous. The above findings for ZnO nanostructures show their potential application in optoelectronic devices.

Vitamin A Fortified Edible Oils and Rice: The Knowledge Level and Consumption Rate among Different Place of Residences in Tangail District, Bangladesh.

A population based cross-sectional study was conducted in four residential areas of Tangail Sadar Upazila, Bangladesh aiming to explore the knowledge about vitamin A fortified edible oils and rice as well as the their consumption rate. A total of 400 participants were randomly selected. Data were collected through face to face interviews. Only 15.0% and 3.5% participants had knowledge about vitamin A fortified edible oils and rice respectively. Precisely vitamin A fortified edible oils knowledge level was found inferior in rural (11.7%) and slum (0%) than urban (48.3%) and semi-urban (40.0%) participants. Knowledge about vitamin A fortified rice was found higher in urban (78.6%) than rural (14.3%) and slum (0%) participants. Most of the participants received knowledge through advertisements (N=45 and N=12 for fortified oils and rice respectively). Maximum participants (62.3%) did not consume vitamin A fortified edible oils and consumption rate in rural (11.8%) and slum (10.2%) areas were poorer than urban (53.5%) and semi-urban (24.4%) areas. Consumption rate of vitamin A fortified rice was found nil in all the study areas. Lack of knowledge about vitamin A fortified rice and oils accounted solely for poor consumption. Knowledge level about vitamin A fortified edible oils and rice and consumption rate were found statistically significantly (p<0.05) with participant's place of residences, education level and monthly income. Finally, this study indicates that the overall knowledge level and consumption rate of vitamin A fortified edible oils and rice is poor especially in rural and slum populations in Tangail Sadar Upazila.

Zoonotic tuberculosis knowledge and practices among cattle handlers in selected districts of Bangladesh.

We assessed zoonotic tuberculosis (zTB) knowledge and prevention and control practices of 404 cattle handlers via a survey in three dairy-intensive districts of Bangladesh. Most respondents were aged 30-49 (52%) and male (95%). Almost all (99%) recognized the important public health burden of tuberculosis in Bangladesh, however, most (58%) had inadequate knowledge about zTB transmission to humans. Inappropriate practices such as: not using protective equipment (98%); smoking, drinking or eating food whilst working with cattle (69%); and sharing the same premises with animals (83%) were identified. Cattle handlers educated at secondary or higher levels were 2.82- (95% CI: 1.59-5.10) and 5.15 times (95% CI: 1.74-15.20) more likely to have adequate knowledge of control and prevention activities compared to those with no formal education. Those who had reared animals for 1-5 years were 2.67 times (95% CI: 1.44-4.91) more likely to have adequate knowledge, compared to those who reared animals for >15 years. Cattle handlers with a monthly incomes of 10,000-20,000 taka were significantly (Odds Ratio = 0.36, 95% CI: 0.14-0.92) less likely to have adequate knowledge compared to those with monthly incomes <10,000 taka. Cattle handlers with high school or higher education were 6.98 times (95% CI: 2.47-19.71) more likely to use appropriate zTB control and prevention practices compared to those without formal education. Those who had reared animals for 1-5 years, 6-10 years and 11-15 years were 2.72- (95% CI: 1.42-5.24), 2.49- (95% CI: 1.29-4.77) and 2.86 times (95% CI: 1.13-7.23) more likely to apply appropriate practices compared to those who reared animals for >15 years. Overall, education, duration of cattle rearing and monthly income predicted zTB knowledge and practices. There is an urgent need to educate those at high-risk of zTB transmission on issues including the handling of infected animals, and general hygiene. A One Health approach, to support the Sustainable Development Goals and the End TB strategy, appears to be the way forward.

Ircinialactams: subunit-selective glycine receptor modulators from Australian sponges of the family Irciniidae.

Screening an extract library of >2500 southern Australian and Antarctic marine invertebrates and algae for modulators of glycine receptor (GlyR) chloride channels identified three Irciniidae sponges that yielded new examples of a rare class of glycinyl lactam sesterterpene, ircinialactam A, 8-hydroxyircinialactam A, 8-hydroxyircinialactam B, ircinialactam C, ent-ircinialactam C and ircinialactam D. Structure-activity relationship (SAR) investigations revealed a new pharmacophore with potent and subunit selective modulatory properties against alpha1 and alpha3 GlyR isoforms. Such GlyR modulators have potential application as pharmacological tools, and as leads for the development of GlyR targeting therapeutics to treat chronic inflammatory pain, epilepsy, spasticity and hyperekplexia.

Behaviour problems in young children in rural Bangladesh.

OBJECTIVE: To determine the prevalence of child behaviour problems reported by parents in rural Bangladesh. METHODS: A total of 4,003 children aged 2-9 years were identified during a population-based survey of 2,231 households. A predetermined sample of 499 was selected, of which health professionals saw 453 (90.8%) for structured physical and neurological examination, standardized testing of cognition and adaptive behaviour and parent report of developmental history and behaviour problems. RESULTS: The prevalence of behaviour impairments was 14.6% (95% CI 11.4, 17.9). The majority involved somatic complaints, including nocturnal enuresis and pica. Problems such as aggression or restlessness were infrequently reported. Behaviour impairments were significantly associated with malnutrition (prevalence ratio 2.1, 95% CI 1.2, 3.6, p < 0.01) and cognitive, motor or seizure disabilities (prevalence ratio 1.8, 95% CI 1.1, 2.9, p < 0.05). CONCLUSIONS: The prevalence and nature of reported behaviour impairments in rural Bangladesh have implications for public health planning and delivery of health services.

Effect of starch 1500 as a binder and disintegrant in lamivudine tablets prepared by high shear wet granulation.

High shear wet granulation is a preferred manufacturing method of tablets. It allowed for rapid production of compressible granulations. The resultant granulation characteristics depend on a combination of formulation properties and processing parameters. Fully pregelatinized starches are currently being used as binders in wet granulated formulations. But due to the gelatinization, much of the disintegration properties are lost. Partially pregelatinized starches (starch 1,500) have a mixture of properties of both native and fully gelatinized starches; made them useful as both a binder and a disintegrant in wet granulated formulations. Starch 1,500 performed as an excellent binder producing a granulation that was compressible and produced lamivudine tablets of improved hardness and friability compared with those prepared with povidone. The formulation of lamivudine tablets with starch 1,500 exceeded the disintegration and dissolution performance of the povidone formulation that utilized a super disintegrant. High shear wet granulation is also well suited for the use of partially pregelatinized starches.

Simultaneous high-performance liquid chromatographic determination of atenolol and amlodipine in pharmaceutical-dosage form.

A simple, rapid and precise method is developed for the quantitative simultaneous determination of atenolol and amlodipine in a combined pharmaceutical-dosage form. The method is based on High Performance Liquid Chromatography (HPLC) on a reversed-phase column, shim-pack CLC, ODS (C18), 4.6 mmx25 cm & 0.5 mm, using a mobile phase of ammonium acetate buffer (the pH was adjusted to 4.5+/-0.05 with glacial acetic acid), acetonitrile and methanol (35::30:35 v/v). The buffer used in the mobile phase contains ammonium acetate in double-distilled water. The chromatographic conditions are- flow rate of 1.5 ml/min, column temperature at 40 degrees C and detector wavelength of 237 nm. Both the drugs were well resolved on the stationary phase and the retention times were around 1.5 minute for atenolol and 3.4 minute for amlodipine. The method was validated and shown to be linear for atenolol and amlodipine. The correlation coefficients for atenolol and amlodipine are 0.999963 and 0.999979, respectively. The relative standard deviations for six replicate measurements in two sets of each drug in the tablets is always less than 2% and mean % error of active recovery not more than +/-1.5%. The method was validated for precision and accuracy. The proposed method was successfully applied to the pharmaceutical dosage forms containing the above-mentioned drug combination without any interference by the excipients.

A Chemogenetic Receptor That Enhances the Magnitude and Frequency of Glycinergic Inhibitory Postsynaptic Currents without Inducing a Tonic Chloride Flux.

The gene transfer-mediated expression of inhibitory ion channels in nociceptive neurons holds promise for treating intractable pain. Chemogenetics, which involves expressing constructs activated by biologically inert molecules, is of particular interest as it permits tunable neuromodulation. However, current chloride-permeable chemogenetic constructs are problematic as they mediate a tonic chloride influx which over time would deplete the chloride electrochemical gradient and reduce inhibitory efficacy. Inflammatory pain sensitization can be caused by prostaglandin E2-mediated inhibition of glycinergic inhibitory postsynaptic currents in spinal nociceptive neurons. We developed a highly conducting (100 pS) inhibitory chemogenetic construct based on a human glycine receptor (α1Y279F,A288G) with high ivermectin sensitivity. When virally infected into spinal neurons, 10 nM ivermectin increased the magnitude and frequency of glycinergic postsynaptic currents without activating a tonic chloride flux. The construct should thus produce analgesia. Its human origin and the well-established biocompatibility of its ligand suggest it may be suited to human use.

Ivermectin-Activated, Cation-Permeable Glycine Receptors for the Chemogenetic Control of Neuronal Excitation.

The ability to control neuronal activation is rapidly advancing our understanding of brain function and is widely viewed as having eventual therapeutic application. Although several highly effective optogenetic, optochemical genetic, and chemogenetic techniques have been developed for this purpose, new approaches may provide better solutions for addressing particular questions and would increase the number of neuronal populations that can be controlled independently. An early chemogenetic neuronal silencing method employed a glutamate receptor Cl- channel engineered for activation by 1-3 nM ivermectin. This construct has been validated in vivo. Here, we sought to develop cation-permeable ivermectin-gated receptors that were either maximally Ca2+-permeable so as to induce neuro-excitotoxic cell death or minimally Ca2+-permeable so as to depolarize neurons with minimal excitotoxic risk. Our constructs were based on the human α1 glycine receptor Cl- channel due to its high conductance, human origin, high ivermectin sensitivity (once mutated), and because pore mutations that render it permeable to Na+ alone or Na+ plus Ca2+ are well characterized. We developed a Ca2+-impermeable excitatory receptor by introducing the F207A/P-2'Δ/A-1'E/T13'V/A288G mutations and a Ca2+-permeable excitatory receptor by introducing the F207A/A-1'E/A288G mutations. The latter receptor efficiently induces cell death and strongly depolarizes neurons at nanomolar ivermectin concentrations.

Antibacterial and cytotoxic compounds from the bark of Cananga odorata.

O-Methylmoschatoline, liriodenine and 3,4-dihydroxybenzoic acid isolated from the barks of Cananga odorata showed antibacterial activities against a number of Gram (+) and Gram (-) bacteria. The compounds also showed antifungal and cytotoxic activities.

Mechanism of action of the insecticides, lindane and fipronil, on glycine receptor chloride channels.

BACKGROUND AND PURPOSE: Docking studies predict that the insecticides, lindane and fipronil, block GABA(A) receptors by binding to 6' pore-lining residues. However, this has never been tested at any Cys-loop receptor. The neurotoxic effects of these insecticides are also thought to be mediated by GABA(A) receptors, although a recent morphological study suggested glycine receptors mediated fipronil toxicity in zebrafish. Here we investigated whether human α1, α1ß, α2 and α3 glycine receptors were sufficiently sensitive to block by either compound as to represent possible neurotoxicity targets. We also investigated the mechanisms by which lindane and fipronil inhibit α1 glycine receptors. EXPERIMENTAL APPROACH: Glycine receptors were recombinantly expressed in HEK293 cells and insecticide effects were studied using patch-clamp electrophysiology. KEY RESULTS: Both compounds completely inhibited all tested glycine receptor subtypes with IC(50) values ranging from 0.2-2 µM, similar to their potencies at vertebrate GABA(A) receptors. Consistent with molecular docking predictions, both lindane and fipronil interacted with 6' threonine residues via hydrophobic interactions and hydrogen bonds. In contrast with predictions, we found no evidence for lindane interacting at the 2' level. We present evidence for fipronil binding in a non-blocking mode in the anaesthetic binding pocket, and for lindane as an excellent pharmacological tool for identifying the presence of ß subunits in αß heteromeric glycine receptors. CONCLUSIONS AND IMPLICATIONS: This study implicates glycine receptors as novel vertebrate toxicity targets for fipronil and lindane. Furthermore, lindane interacted with pore-lining 6' threonine residues, whereas fipronil may have both pore and non-pore binding sites.

High Throughput Techniques for Discovering New Glycine Receptor Modulators and their Binding Sites.

The inhibitory glycine receptor (GlyR) is a member of the Cys-loop receptor family that mediates inhibitory neurotransmission in the central nervous system. These receptors are emerging as potential drug targets for inflammatory pain, immunomodulation, spasticity and epilepsy. Antagonists that specifically inhibit particular GlyR isoforms are also required as pharmacological probes for elucidating the roles of particular GlyR isoforms in health and disease. Although a substantial number of both positive and negative GlyR modulators have been identified, very few of these are specific for the GlyR over other receptor types. Thus, the potential of known compounds as either therapeutic leads or pharmacological probes is limited. It is therefore surprising that there have been few published studies describing attempts to discover novel GlyR isoform-specific modulators. The first aim of this review is to consider various methods for efficiently screening compounds against these receptors. We conclude that an anion sensitive yellow fluorescent protein is optimal for primary screening and that automated electrophysiology of cells stably expressing GlyRs is useful for confirming hits and quantitating the actions of identified compounds. The second aim of this review is to demonstrate how these techniques are used in our laboratory for the purpose of both discovering novel GlyR-active compounds and characterizing their binding sites. We also describe a reliable, cost effective method for transfecting HEK293 cells in single wells of a 384-well plate using nanogram quantities of plasmid DNA.

Toxicological studies of two compounds isolated from Loranthus globosus Roxb.

The sub-acute toxicities of two compounds 3,4-dimethoxycinnamyl alcohol (1) and 3,4,5-trimethoxycinnamyl alcohol (2) isolated from the plant Loranthus globosus Roxb were studied on long Evan's rats. The studies included the gross general observation such as changes in body weight, haematological profiles [total count of Red Blood Cells (RBC) and White Blood Cells (WBC), differential count of WBC, platelet count and Haemoglobin (Hb)%], biochemical parameters of blood [Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT), Serum Alkaline Phosphatase (SALP), urea and creatinine) and histopathology of the liver, kidney, heart and lung of both control and experimental groups of rats. The changes in haematological and biochemical parameters were statistically not significant after the administration of compounds 1 and 2 in a dose of 300 microg/rat/day for consecutive 14 days. No abnormality was found in the histopathology of the liver, kidney, heart and lung in the experimental groups of rats following same dose when compared with control group. This preliminary study suggests that the isolated compounds may be used safely for clinical trial.