Data analytics in control and operation of municipal wastewater treatment plants: qualitative analysis of needs and barriers.

This study aims to identify barriers and needs for the application of data analytics in municipal wastewater treatment. The study was conducted through a series of interviews with stakeholders involved in instrumentation, control, and automation of wastewater treatment plants. Opportunities and limitations observed by different stakeholders were assessed with a thematic analysis. Thematic analysis enabled a broader consideration of social and organizational aspects related to process control, operation, and maintenance. Identified key barriers for applying data analytics included laborious instrumentation maintenance, unstable control loops, and deficient customization of digital tools for users at wastewater treatment plants. Development needs include easier data processing tools, quality assurance of instrumentation, and controller tuning. Results indicate that the perceived potential of data analytics is highly dependent on the performance of underlying physical and digital systems, as well as the control strategies and operating environment of the plant. Despite the barriers, users and developers see many potential applications for data analytics and expect them to have a central role in the control and operation of wastewater treatment plants in the future.

Prescribing antimicrobial agents for dogs and cats via university pharmacies in Finland--patterns and quality of information.

The aim of our study was to evaluate antimicrobial use in dogs and cats in Finland. Information on veterinary prescriptions was gathered from University Pharmacies (n=17) over a one-month period, April 2001. A total of 2719 prescriptions for veterinary use were delivered, of which the majority were for dogs (70%, n=1898) and cats (14%, n=384). The most prescribed therapy group was per-oral antimicrobial agents (53%, n=1449), of which 16% (n=237) were medicines approved for humans. The most commonly used substances for dogs and cats were betalactams, 66% and 78%, respectively. The proportion of fluoroquinolones was 3-5%. The average duration of the treatment periods was 10 days with the exception of treatment of cats with macrolide-lincosamides, where the mean period was 20 days. Indication was mentioned only in 37% of the prescriptions.

Construction of gene-targeting vectors: a rapid Mu in vitro DNA transposition-based strategy generating null, potentially hypomorphic, and conditional alleles.

Gene targeting into mammalian genomes by means of homologous recombination is a powerful technique for analyzing gene function through generation of transgenic animals. Hundreds of mouse strains carrying targeted alleles have already been created and recent modifications of the technology, in particular generation of conditional alleles, have extended the usefulness of the methodology for a variety of special purposes. Even though the standard protocols, including the construction of gene-targeting vector plasmids, are relatively straightforward, they typically involve time-consuming and laborious gene mapping and/or sequencing steps. To produce various types of gene-targeting constructions rapidly and with minimum effort, we developed a strategy, that utilizes a highly efficient in vitro transposition reaction of phage Mu, and tested it in a targeting of the mouse Kcc2 gene locus. A vast number and different types of targeting constructions can be generated simultaneously with little or no prior sequence knowledge of the gene locus of interest. This quick and efficient general strategy will facilitate easy generation of null, potentially hypomorphic, and conditional alleles. Especially useful it will be in the cases when effects of several exons within a given gene are to be studied, a task that necessarily will involve generation of multiple constructions. The strategy extends the use of diverse recombination reactions for advanced genome engineering and complements existing recombination-based approaches for generation of gene-targeting constructions.

An efficient DNA sequencing strategy based on the bacteriophage mu in vitro DNA transposition reaction.

A highly efficient DNA sequencing strategy was developed on the basis of the bacteriophage Mu in vitro DNA transposition reaction. In the reaction, an artificial transposon with a chloramphenicol acetyltransferase (cat) gene as a selectable marker integrated into the target plasmid DNA containing a 10.3-kb mouse genomic insert to be sequenced. Bacterial clones carrying plasmids with the transposon insertions in different positions were produced by transforming transposition reaction products into Escherichia coli cells that were then selected on appropriate selection plates. Plasmids from individual clones were isolated and used as templates for DNA sequencing, each with two primers specific for the transposon sequence but reading the sequence into opposite directions, thus creating a minicontig. By combining the information from overlapping minicontigs, the sequence of the entire 10,288-bp region of mouse genome including six exons of mouse Kcc2 gene was obtained. The results indicated that the described methodology is extremely well suited for DNA sequencing projects in which considerable sequence information is on demand. In addition, massive DNA sequencing projects, including those of full genomes, are expected to benefit substantially from the Mu strategy.

Retarded growth and deficits in the enteric and parasympathetic nervous system in mice lacking GFR alpha2, a functional neurturin receptor.

Glial cell line-derived neurotrophic factor (GDNF) and a related protein, neurturin (NTN), require a GPI-linked coreceptor, either GFR alpha1 or GFR alpha2, for signaling via the transmembrane Ret tyrosine kinase. We show that mice lacking functional GFR alpha2 coreceptor (Gfra2-/-) are viable and fertile but have dry eyes and grow poorly after weaning, presumably due to malnutrition. While the sympathetic innervation appeared normal, the parasympathetic cholinergic innervation was almost absent in the lacrimal and salivary glands and severely reduced in the small bowel. Neurite outgrowth and trophic effects of NTN at low concentrations were lacking in Gfra2-/- trigeminal neurons in vitro, whereas responses to GDNF were similar between the genotypes. Thus, GFR alpha2 is a physiological NTN receptor, essential for the development of specific postganglionic parasympathetic neurons.

Does plasma L-arginine level play a role in regulation of blood pressure during exercise and autonomic nervous function?

Little is known of L-arginine's role in autonomic nervous regulation and physiological responses to dynamic exercise. We assessed heart rate and blood pressure during a maximal bicycle ergometer test and heart rate variability at rest in 15 healthy male volunteers, age 22-38 years. Venous blood samples for plasma L-arginine measurements were taken when subjects were sitting at rest before and at the end of exercise. The autonomic nervous function was assessed with time and frequency domain analysis of heart rate variability. Plasma L-arginine level decreased during maximal exercise from 71.4 mumol l-1 to 51.0 mumol l-1 (P < 0.0001) for all subjects studied. The systolic blood pressure during the maximal exercise test was inversely correlated with plasma L-arginine level at rest (r = -0.70, P < 0.01). Normalized low frequency band of power spectral analysis of heart rate variability correlated with L-arginine level at rest (r = 0.66, P < 0.01). In conclusion, plasma L-arginine level decreased in physical exercise, and plasma L-arginine level at rest was positively associated with the sympathetic component of power spectral analysis of heart rate variability at rest, and inversely with systolic blood pressure during physical exercise.

Glutamate-stimulated ROS production in neuronal cultures: interactions with lead and the cholinergic system.

Oxidative stress may be an important factor in several pathological brain conditions. A contributing factor in many such conditions is excessive glutamate release, and subsequent glutamatergic neuronal stimulation, that causes increased production of reactive oxygen species (ROS), oxidative stress, excitotoxicity and neuronal damage. Glutamate release is also associated with illnesses such as Alzheimer's disease, stroke, and brain injury. Glutamate may interact with an environmental toxin, lead, and this interaction may result in neuronal damage. Glutamate-induced ROS production is greatly amplified by lead in cultured neuronal cells. Alterations in protein kinase C (PKC) activity seem to be important both for glutamate-induced ROS production, and for the amplification of glutamate-induced ROS production by lead. It is possible that the neurotoxic effects of lead are amplified through glutamate-induced neuronal excitation. Cholinergic stimulation can also trigger ROS production in neuronal cells. PKC seems to play a key-role also in cholinergic-induced ROS production superoxide anion being the primary reactive oxygen species. There seems to be a close relationship between the responses of cholinergic muscarinic and glutamatergic receptors because glutamate receptor antagonists inhibit cholinergic-induced activation of human neuroblastoma cells. Glutamatergic neuronal stimulation may be a common final pathway in several brain conditions in which oxidative stress and ensuing excitotoxicity plays a role.

Interactions of excitatory neurotransmitters and xenobiotics in excitotoxicity and oxidative stress: glutamate and lead.

Increased glutamate release is associated with serious neurological disorders such as epilepsy, stroke, Alzheimer's disease and other brain injuries. Excessive glutamate release and subsequent glutamatergic neuronal stimulation increase the production of reactive oxygen species (ROS), which in turn induce oxidative stress, excitotoxicity and neuronal damage. A number of studies have shown that co-exposure of neuronal cells to glutamate, and an environmental toxin, lead, can greatly amplify glutamate excitotoxicity and cell death through apoptosis or necrosis. Even though the mechanisms of excitotoxicity or those of glutamate-lead interactions have not been exhaustively delineated, there is ample evidence to suggest that increased production of ROS may play an important role in both events. Subsequently, increased DNA binding of redox-regulated transcription factors, NF-kappaB and AP-1, seems to be associated with these events. Induction of an immediate early gene, c-fos, is seen in neuronal cells exposed to glutamate or lead. Immediate early genes are important in regulating the expression of other neuronal genes; Elevated expressions of the genes encoding Hsp70 or cyclo-oxygenase-2 seem to be involved in the apoptosis or necrosis induced by glutamate, and may be associated with induction of several of the genes in cells exposed to lead, or to the glutamate-lead combination. Further studies are required to clarify the mechanisms of glutamate-lead neurotoxicity.