Synthetic α-Conotoxin Mutants as Probes for Studying Nicotinic Acetylcholine Receptors and in the Development of Novel Drug Leads

α-Conotoxins are peptide neurotoxins isolated from venomous marine cone snails that are potent and selective antagonists for different subtypes of nicotinic acetylcholine receptors (nAChRs). As such, they are valuable probes for dissecting the role that nAChRs play in nervous system function. In recent years, extensive insight into the binding mechanisms of α-conotoxins with nAChRs at the molecular level has aided in the design of synthetic analogs with improved pharmacological properties. This review examines the structure-activity relationship studies involving α-conotoxins as research tools for studying nAChRs in the central and peripheral nervous systems and their use towards the development of novel therapeutics

alpha-conotoxin EpI, a novel sulfated peptide from Conus episcopatus that selectively targets neuronal nicotinic acetylcholine receptors

We have isolated and characterized ol-conotoxin EpI, a novel sulfated peptide from the venom of the molluscivorous snail, Conus episcopatus, The peptide was classified as an cy-conotoxin based on sequence, disulfide connectivity, and pharmacological target. EpI has ho mology to sequences of previously described cu-conotoxins, particularly PnIA, PnIB, and ImI, However, EpI differs from previously reported conotoxins in that it has a sulfotyrosine residue, identified by amino acid analysis and mass spectrometry, Native EpI was shown to coelute with synthetic EpI, The peptide sequence is consistent with most, but not all, recognized criteria for predicting tyrosine sulfation sites in proteins and peptides, The activities of synthetic EpI and its unsulfated analogue [Tyr(15)]EpI were similar. Both peptides caused competitive inhibition of nicotine action on bovine adrenal chromaffin cells (neuronal nicotinic ACh receptors) but had no effect on the rat phrenic nerve-diaphragm (muscle nicotinic ACh receptors), Both EpI and [Tyr(15)]EpI partly inhibited acetylcholine-evoked currents in isolated parasympathetic neurons of rat intracardiac ganglia, These results indicate that EPI and [Tyr(15)]EpI selectively inhibit alpha 3 beta 2 and alpha 3 beta 4 nicotinic acetylcholine receptors

An ecological assessment of seven major lakes in the Philippines

Abstract only.An ecological assessment was done on the seven major lakes in the Philippines, namely; Laguna de Bay, Taal, Naujan, Lanao, Mainit, Buluan, and Bato. The assessment was based largely on secondary data and some primary data. The ecological parameters considered for each lake were focused on published information as well as those unpublished but were made accessible to the author by researchers and agencies of government. The different lakes are classified into three ecological categories, namely; critically degraded, degraded, threatened, and underdeveloped. Where adequate data on a lake is available, emphasis is also given on the examination of the nature of land use, lake development measures, climatic variations in terms of rainfall, and ecological conditions in the lake as they affect fish production. Finally, lake specific and general recommendations are forwarded for management purposes

Agroindustrial waste products as sources of cheap substrates for algal single-cell protein production

Four types of agroindustrial waste products were tested for their suitability as substrates for Chlorella single-cell protein production. Based on cell density on day 7, unsterilized rice straw filtrate, Tris-buffered inorganic medium (control), unsterilized slop (1%) and unsterilized rice hull filtrate were suitable culture media. Dry weight yield after two weeks did not vary among media. Gross protein content of algae was highest in rice straw with or without sterilization. Substrates for algal SCP production are assessed on the basis of their nutrient content, pH, and hygienic acceptability

Structure of PBP-A from Thermosynechococcus elongatus, a Penicillin-Binding Protein Closely Related to Class A β-Lactamases

Molecular evolution has always been a subject of discussions, and researchers are interested in understanding how proteins with similar scaffolds can catalyze different reactions. In the superfamily of serine penicillin-recognizing enzymes, D-alanyl-D-alanine peptidases and β-lactamases are phylogenetically linked but feature large differences of reactivity towards their respective substrates. In particular, while β-lactamases hydrolyze penicillins very fast, leading to their inactivation, these molecules inhibit D-alanyl-D-alanine peptidases by forming stable covalent penicilloyl enzymes. In cyanobacteria, we have discovered a new family of penicillin-binding proteins (PBPs) presenting all the sequence features of class A β-lactamases but having a six-amino-acid deletion in the conserved Ω-loop and lacking the essential Glu166 known to be involved in the penicillin hydrolysis mechanism. With the aim of evolving a member of this family into a β-lactamase, PBP-A from Thermosynechococcus elongatus has been chosen because of its thermostability. Based on sequence alignments, introduction of a glutamate in position 158 of the shorter Ω-loop afforded an enzyme with a 50-fold increase in the rate of penicillin hydrolysis. The crystal structures of PBP-A in the free and penicilloylated forms at 1.9 Å resolution and of L158E mutant at 1.5 Å resolution were also solved, giving insights in the catalytic mechanism of the proteins. Since all the active-site elements of PBP-A-L158E, including an essential water molecule, are almost perfectly superimposed with those of a class A β-lactamase such as TEM-1, the question why our mutant is still 5 orders of magnitude less active as a penicillinase remains and our results emphasize how far we are from understanding the secrets of enzymes. Based on the few minor differences between the active sites of PBP-A and TEM-1,mutations were introduced in the L158E enzyme, but while activities on D-Ala-D-Ala mimicking substrates were severely impaired, further improvement in penicillinase activity was unsuccessful

Socio-economic study of two major Metro Manila esteros

Two major Manila esteros namely Estero de San Miguel and Estero de Quiapo, were studied to determine their socio-economic status. Socio-economic study revealed that attitude and actions of stakeholders to their esteros were influenced primarily by their economic status, educational and cultural background. Survey also showed the awareness of the communities on the problems of esteros and their willingness to cooperate in the rehabilitation of their polluted canals. Institutional initiatives such as environmental education and community-based programs were looked into as possible key for the effective rehabilitation and attainment of sustainability of Manila esteros