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At least 865 records · Page 48Linked to original sources
PubMed1990

Indications for selective coupling to phosphoinositide hydrolysis or to adenylate cyclase inhibition by endogenous muscarinic receptor subtypes M3 and M4 but not by M2 in tumor cell lines.

The muscarinic receptor subtype mRNAs expressed in cell lines were determined by Northern blot analysis. The biochemical responses of the muscarinic receptors in these cell lines (phosphoinositide hydrolysis and cAMP levels) were studied and correlated to the corresponding muscarinic receptor subtype as determined by mRNA expression. PC12 cells that expressed M4 subtype mRNA exhibited muscarinically dependent adenylate cyclase inhibition, whereas C6 and SK-N-SH cells expressing M3 subtype mRNA exhibited muscarinically dependent phosphoinositide hydrolysis. IMR-32 cells (M2 subtype mRNA) exhibited both muscarinically dependent phosphoinositide hydrolysis and adenylate cyclase inhibition. These results suggest that endogenous M3 and M4 receptor subtypes are selectively coupled to phosphoinositide hydrolysis and adenylate cyclase inhibition, respectively, whereas the M2 receptor subtype is coupled to both responses.

Adenylyl Cyclases↗
PubMed1991

Regional and laminar distributions of alpha 1-adrenoceptors and their subtypes in human and rat hippocampus.

The distributions of the alpha 1-adrenoceptor and its subtypes (alpha 1A and alpha 1B) in human and rat hippocampus are analysed by quantitative receptor autoradiography. alpha 1-Adrenoceptors are labelled by [3H]prazosin. The alpha 1A subtype is visualized by [3H]prazosin after irreversible blockade of alpha 1B adrenoceptors with chloroethylclonidine or directly by [3H]5-methyl-urapidil. The alpha 1B subtype is investigated by [3H]prazosin binding in the presence of the alpha 1A antagonist 5-methyl-urapidil. Considerable differences in the regional and laminar patterns of alpha 1-adrenoceptors are found between rat and human hippocampi. The rat hippocampus is characterized by a low overall density and a rather homogeneous regional and laminar distribution. This is in contrast to the human pattern, which shows a much higher overall level of alpha 1 receptor density and a restriction of alpha 1 receptors to the CA3 region of Ammon's horn and the dentate gyrus. Moreover, alpha 1A and alpha 1B receptors of the human hippocampus are differentially distributed with the alpha 1A subtype concentrated in the hilus and lucidum layer of CA3, and the alpha 1B subtype concentrated in the molecular layer of the dentate gyrus. Additionally, the distribution of alpha 1 receptors is compared with the distribution of 5-hydroxytryptamine 1A receptors. The subtype specific pattern is correlated with the distribution of glutamatergic systems in the human (but not in the rat) hippocampus. alpha 1A Receptor localization coincides with the target area of the mossy fibre system, and alpha 1B receptors are preferentially localized in the target area of the hippocampal associational fibres and partly of the perforant pathway. This result points to possible interactions between noradrenaline- and glutamate-mediated neurotransmission differentiated by topographically segregated alpha 1-adrenoceptor subtypes.

Adrenergic alpha-Antagonists↗
PubMed1993

Expression of multiple alpha adrenergic receptor subtype messenger RNAs in the adult rat brain.

Multiple subtypes of alpha adrenergic receptors with CNS expression (alpha 1A, alpha 1B, alpha 2A and alpha 2C) have been identified through pharmacological and molecular biological means. To characterize the localization of these subtypes and attempt to correlate subtype expression with physiological significance, the expression of the mRNAs encoding the alpha 1A, alpha 1B, alpha 2A and alpha 2C adrenergic receptor subtypes was examined in the adult rat brain by in situ hybridization histochemistry. Each subtype demonstrated a unique pattern of distribution, with the alpha 1 adrenergic receptors more restricted in their distribution and the alpha 2 receptors more widespread. The alpha 1A was primarily localized in the olfactory bulb, intermediate layers of the cortex, the hippocampus and the reticular nucleus of the thalamus. The alpha 1B was expressed in intermediate and deep layers of the cortex, thalamus, hippocampus, dorsal raphe and cerebellum. Although the alpha 2A message was relatively low in abundance, it was identified in the olfactory bulb, cortex, hippocampus, locus coeruleus, pons and cerebellum. The alpha 2C messenger RNA was localized in the cortex (particularly cingulate), hippocampus, caudoputamen, pons and cerebellum. Multiple alpha adrenergic receptor subtypes have significant sequence homology and similar pharmacologic properties; however, they each possess a unique pattern of messenger RNA distribution throughout the brain. The multiplicity of subtypes of alpha adrenergic receptors in specific brain regions may dictate the physiological and pharmacological responses to catecholamines.

Animals↗
PubMed1996

GABAA receptor subtype changes in the substantia nigra of the rat following quinolinate lesions in the striatum: a correlative in situ hybridization and immunohistochemical study.

This study investigates the pattern of distribution of GABAA receptor subunit subtypes in the substantia nigra of the rat using in situ hybridization techniques and immunohistochemistry at the light microscopic level following unilateral quinolinate lesions in the striatum. The main purpose of this study was to first identify the variety and regional distribution of GABAA receptor subtype messenger RNAs in the normal substantia nigra and, second, to determine if this pattern and level of expression of GABAA receptor subtypes in the substantia nigra is affected following quinolinate-induced degeneration of the GABAergic striatonigral projection neurons. The study is based on a comparison of adjacent sections using: (i) in situ hybridization and oligonucleotide probes selective for 13 of the GABAA receptor subunits; and (ii) immunohistochemistry and antibodies specific to three protein subunits of the GABAA receptor complex. The results show that the GABAA receptor in the normal substantia nigra pars reticulata has a molecular configuration comprising of the alpha 1, beta 2, and gamma 2 subtypes and that following quinolinate lesions of the striatum the subtype configuration of the GABAA receptors remains unaltered, but that there is a marked increase in the level of expression of the alpha 1, beta 2 and gamma 2 subtypes. In confirmation of these findings, the immunohistochemical results show increased immunoreactivity for the alpha 1, beta 2,3 and gamma 2 GABAA receptor subtypes in the substantia nigra following degeneration of GABAergic striatonigral neurons. The details of these findings are discussed with reference to previous studies and with regard to the implications that these results may have for specific GABAergic neurodegenerative diseases of the human basal ganglia, such as Huntington's disease.

Animals↗
PubMed1996

Expression of alpha 2-adrenergic receptor subtypes in the mouse brain: evaluation of spatial and temporal information imparted by 3 kb of 5' regulatory sequence for the alpha 2A AR-receptor gene in transgenic animals.

The present studies characterize the expression of the alpha 2A, alpha 2B and alpha 2C adrenergic receptor subtypes via in situ hybridization analysis of messenger RNA expression in the adult mouse brain, as well as the pattern of expression of alpha 2A adrenergic receptor messenger RNA at embryonic day E9.5, the earliest time for detection of the messenger RNA encoding this receptor subtype. alpha 2A adrenergic receptor messenger RNA is highly expressed in the sixth layer of the cortex and the locus coeruleus, alpha 2B adrenergic receptor messenger RNA predominantly in the thalamus and in the Purkinje layer of the cerebellum, and alpha 2C adrenergic receptor messenger RNA in the putamen caudate region of the mouse brain. Both alpha 2A and alpha 2C adrenergic receptor messenger RNA demonstrate strong expression in the amygdaloid complex, hypothalamus, olfactory system and the hippocampal formation. To develop a molecular understanding of the unique cellular expression of messenger RNA encoding the alpha 2A adrenergic receptor subtype, 2.83 kb of the upstream regulatory sequence for the alpha 2A adrenergic receptor gene was fused to the LacZ gene as a reporter gene and expression of beta-galactosidase activity was assessed in transgenic offspring. Although the spatial expression of the transgene in the adult brain often overlaps that for the endogenous alpha 2A adrenergic receptor, both ectopic expression and the absence of appropriate expression were noted; in contrast five of the six lines show temporal expression characteristic of the endogenous alpha 2A adrenergic receptor gene. The present studies provide the first characterization of messenger RNA localization for the three alpha 2 adrenergic receptor subtypes in the mouse CNS. Because the functional roles of the prazosin-sensitive alpha 2B adrenergic receptor and alpha 2C adrenergic receptor subtypes have been masked in most earlier physiological and pharmacological analyses of alpha 2 adrenergic receptor function, identifying the multiple loci alpha 2 adrenergic receptor subtype expression is an important prelude to understanding the functional roles of these three independent receptor populations in the mouse CNS. The findings in the transgenic animals indicating that approximately 3 kb of regulatory sequence has imparted faithful temporal but not spatial expression for the alpha 2A adrenergic receptor gene suggest that additional regulatory information is necessary for appropriate cell specific expression of messenger RNA for the alpha 2A adrenergic receptor subtype.

Amino Acid Sequence↗
PubMed1995

Cross-protection experiments in pigs vaccinated with Actinobacillus pleuropneumoniae subtypes 1A and 1B.

Cross-protection experiments were conducted to determine whether antigenic differences located within the lipopolysaccharides (LPS) of Actinobacillus pleuropneumoniae subtypes 1A and 1B were important with respect to the efficacy of whole cell, formalin-inactivated bacterins. Based on clinical signs, lung lesions scores and mortality rates, pigs immunized with A. pleuropneumoniae subtype 1A were partially protected against severe challenge with both subtypes 1A and 1B. In contrast, 1B vaccinated pigs were not protected against severe challenge with subtype 1A but were partially protected against 1B challenge. Cross-reactive serum antibody levels were measured with an ELISA using outer membranes of subtype 1A or 1B as the coating antigen. Serum antibodies were detected against both subtypes within 2 weeks after the first immunization. Antibody levels increased with time and were generally higher against the homologous subtype coating antigen. We conclude that antigenic variation within a capsular serotype, due to antigenic variation within LPS, can result in the failure of whole cell bacterins to provide protection against challenge with the same capsular serotype. This lack of cross-protection within a capsular serotype provides partial explanation for vaccination failures observed under field conditions.

Actinobacillus Infections↗
PubMedDate not supplied

Application of genomic DNA subtyping by pulsed field gel electrophoresis and restriction enzyme analysis of plasmid DNA to characterize methicillin-resistant Staphylococcus aureus from two nosocomial outbreaks.

Pulsed-field gel electrophoresis (PFGE) and restriction enzyme analysis of plasmid DNA (REAP) were applied to study the epidemiologic relationship among methicillin-resistant Staphylococcus aureus (MRSA) isolates from outbreaks in two hospitals in São Paulo, Brazil: 82 MRSA isolates, 73 from a university hospital and nine from a general adult intensive care unit of a private hospital, were collected from 62 patients: 95% of the MRSAs were also resistant to gentamicin and ciprofloxacin. REAP subtyping of both collections identified six different subtypes: 55 (72.6%) MRSAs from the university hospital and nine isolates from the private hospital shared the same epidemic REAP subtype. Discrimination by restriction of genomic DNA with Sma I followed by PFGE enabled the identification of 14 DNA subtypes. Based on the combined REAP-genomic DNA subtype, the predominant subtype in the university hospital was A/A (44 isolates) whereas the epidemic subtype in the private hospital was A/M (seven isolates). The application of two typing methods showed better discrimination among MRSAs than did either method alone.

Bacterial Typing Techniques↗
PubMed1993

Use of recombinant human alpha 2-adrenoceptors to characterize subtype selectively of antagonist binding.

Cloning of the genes encoding three subtypes of human alpha 2-adrenoceptors allows the separate heterologous expression of each subtype. We have generated stably transfected Shionogi S115 mouse mammary tumour cell lines expressing the human alpha 2-adrenoceptor subtypes alpha 2-C10, alpha 2-C2, and alpha 2-C4 at densities of 0.2-7 pmol/mg total cellular protein. Binding of [3H]rauwolscine was inhibited by co-incubation of S115 cell homogenates with ten alpha 2-adrenoceptor antagonists and oxymetazoline, a partial agonist known to discriminate the receptor subtypes. Other useful agents for discrimination of subtypes were prazosin, chlorpromazine, phentolamine, and yohimbine. The most sensitive indices for differences between the three subtypes were the binding inhibition coefficient (Ki) ratios chlorpromazine/oxymetazoline (alpha 2-C10: 202; alpha 2-C2: 0.004; alpha 2-C4: 0.8), prazosin/oxymetazoline (430; 0.03; 0.5) and chlorpromazine/atipamezole (1612; 5.8; 77). Correlation analysis between our results for human-type receptors and published data for their rat alpha 2-adrenoceptor homologues demonstrated excellent general agreement, with some interspecies differences in the affinity of rauwolscine, phentolamine and oxymetazoline. The use of recombinant human receptors produced in stably transfected cell lines should facilitate the development of new, subtype-selective alpha 2-adrenoceptor ligands.

Adrenergic alpha-Antagonists↗
PubMed1994

Expression of alpha 1-adrenoceptor subtypes in rat tissues: implications for alpha 1-adrenoceptor classification.

We report here the mapping of the mRNA distribution of three different alpha 1-adrenoceptor subtypes (alpha 1b, alpha 1c and alpha 1d) in various rat tissues. cDNA fragments covering the region from the fifth to seventh putative transmembrane spanning domains of these three alpha 1-adrenoceptor subtypes were generated from rat hippocampus using reverse transcription coupled to polymerase chain reaction (PCR). These three alpha 1-adrenoceptor cloned cDNA fragments were then used as subtype-selective cDNA probes in Northern blot analysis. Of the three specific DNA probes only the rat alpha 1b-adrenoceptor probe hybridized to mRNA of rat liver. The rat alpha 1c-adrenoceptor probe hybridized to a mRNA species of 3.7 kb in tissues that have been reported to contain the classical pharmacologically-defined alpha 1A-adrenoceptor such as hippocampus, vas deferens, lung and salivary gland. Also, a major mRNA transcript of 2.7 kb was detected in hippocampus, vas deferens and lung, using the rat alpha 1d-adrenoceptor probe. In addition, pharmacological characterization of [3H]prazosin binding to three stably transfected mammalian cell-lines expressing one of the three alpha 1-adrenoceptor subtypes cloned to date (namely, alpha 1b--of the hamster smooth muscle DDT1-MF2 cell-line, the bovine brain alpha 1c--and the rat cerebral cortical alpha 1d-adrenoceptors) was performed. Of the three cloned alpha 1-adrenoceptor subtypes that alpha 1c-adrenoceptor showed a similar pharmacological profile to that of the classical alpha 1A-adrenoceptor of rat salivary gland. Our data on the pharmacological profile and expression pattern of the alpha 1c-adrenoceptor indicate, in contrast to earlier claims (Schwinn et al., J. Biol. Chem. 265, 1990), that this subtype is in fact the classical pharmacologically-defined alpha 1A-adrenoceptor subtype.

Amino Acid Sequence↗
PubMed2004

Pharmacological properties of serotonin receptor subtypes mediating contraction of bovine inferior alveolar arteries.

OBJECTIVE: To characterise the 5-hydroxytryptamine (5-HT) receptor subtypes mediating contraction of the inferior alveolar artery. Additionally, to determine the role of cyclooxygenase products, nitric oxide, endothelium, monoamine oxidase and 5-HT uptake in modulating contraction of inferior alveolar arteries to 5-HT. METHODS: Contractile responses to 5-HT were examined in vitro using ring segments of bovine inferior alveolar arteries. Affinity constants (K(B)'s) of subtype-selective 5-HT receptor antagonists were determined to characterise the 5-HT receptor-subtypes causing contraction of inferior alveolar arteries. RESULTS: In 100 nM ketanserin or 30 nM spiperone, 5-HT caused a biphasic contraction best-fit by a two-site curve model, where one site was antagonist-sensitive and the other site antagonist-insensitive. 5-HT(2A) receptor-subtype selective antagonists, ketanserin and spiperone, blocked 5-HT induced contraction with K(B)'s of 1.0 and 0.16 nM, respectively. RS102221 (5-HT(2C) selective) and (S)-WAY100135 (5-HT(1A) selective) blocked 5-HT stimulated contraction with low affinities (K(B)'s=100 nM and 330 nM, respectively). GR55562, a 5-HT(1) receptor subtype antagonist with a reported affinity of 500 nM at the 5-HT(1D) receptor subtype, blocked 5-HT induced contraction with a K(B) of 470 nM. Cylooxygenase inhibition with 50 microM ibuprofen caused a 44% increase in maximal contraction to 5-HT; whereas, nitric oxide inhibition with N(G)-nitro-L-arginine, endothelium removal or inhibition of 5-HT uptake and monoamine oxidase with imipramine and iproniazid, respectively, did not affect 5-HT contraction. CONCLUSIONS: Both 5-HT(2A) and 5-HT(1D/1B) receptor subtypes mediate 5-HT induced contraction of the bovine inferior alveolar artery. 5-HT stimulated contraction of the inferior alveolar artery is modulated by a vasodilator prostaglandin.

Alveolar Process↗
PubMed2004

Modelling extracellular domains of GABA-A receptors: subtypes 1, 2, 3, and 5.

GABA is the main inhibitory neurotransmitter in the mammalian central nervous system. When GABA binds to the ubiquitous GABA-A receptors on neurons, chloride channels are activated leading to a rapid increase in chloride conductance that depresses excitatory depolarization. The GABA-A receptors are targets for many clinically important drugs, such as the benzodiazepines, general anaesthetics, and barbiturates. All of these drugs enhance the chloride current activated by GABA. Of the GABA-A receptor family, the subtype 2 is critical for the treatment of anxiety spectrum disorders. To avoid unwanted side effects, it is necessary to find highly selective drugs that interact only with subtype 2 but not with the related receptors such as subtypes 1, 3, and 5. To realize such a goal, it is important to have not only the 3D (dimensional) structure of subtype 2 but also the 3D structures of subtypes 1, 3, and 5. In this study, the 3D structures of all the four subtypes of GABA-A receptors have been derived. The computer-modeled heteropentameric structures bear the following features: (1) each of the five subunits in the pentamer has an intrachain disulfide bond, a hallmark of ligand-gated pentameric channels; (2) those residues which are sensitive to the binding of the benzodiazepine site ligands are grouped around the alpha1,2,3,5/gamma2 interfaces; and (3) those residues which are sensitive to the binding of GABA molecules are grouped around the alpha1,2,3,5/beta2 interfaces. All these findings are fully consistent with experimental observations. Meanwhile, for those sensitive or key residues, a close look at their subtle difference among the four subtypes has been provided through a highlighted superposition picture. In addition to providing the atomic coordinates, the predicted structures have further clarified some ambiguities that could not been uniquely determined by the existing experimental data, such as the directionality of the subunit arrangement in the heteropentamers. The 3D models may provide a reasonable structural frame or footing for designing highly selective drugs. The present models might be also useful in understanding the basic mechanism of operation of the GABA-A receptors, stimulating novel strategies for developing more specific drugs and better treatments.

Amino Acid Sequence↗
PubMed2004

alpha2-adrenergic receptor subtypes - novel functions uncovered in gene-targeted mouse models.

The family of adrenergic receptors contains nine different subtypes of G protein-coupled receptors which mediate the biological effects of adrenaline and noradrenaline. With few exceptions, the full therapeutic potential of subtype-selective therapy has not yet been explored for the group of adrenergic receptors. In the absence of sufficiently subtype-selective ligands which can distinguish between individual receptor subtypes of the adrenergic family, gene-targeted mouse models with deletions in these receptor genes have recently been generated and characterized. These genetic mouse models have helped to assign specific pharmacological effects of alpha(2)-receptor agonists or antagonists to individual receptor subtypes. However, some unexpected and novel functions of alpha(2)-adrenergic receptors were also uncovered in these mouse models: Presynaptic control of catecholamine release from adrenergic nerves in the central and sympathetic nervous system may be regulated by three different alpha(2)-receptor subtypes, alpha(2A), alpha(2B), and alpha(2C). A similar feedback loop also controls the release of catecholamines from the adrenal gland. alpha(2B)-receptors are not only involved in regulating vascular tone in the adult organism, but they are essential for the development of the vascular system of the placenta during prenatal development. The challenge will now be to generate strategies to identify whether the findings obtained in gene-targeted mice may predict the action of receptor subtype-selective drugs in humans.

Adenylyl Cyclases↗
PubMed2005

Effects of the xenoestrogen bisphenol A in diencephalic regions of the teleost fish Coris julis occur preferentially via distinct somatostatin receptor subtypes.

The xenoestrogen bisphenol A, a contaminant used in the manufacturing of polymers for many consumer products, has been shown to mimic estrogenic actions. This xenoestrogen regulates secretion and expression of pituitary lactotrophs plus morphological and structural features of estrogen target tissues in rodents. Recently, ecological hazards produced by bisphenol A have drawn interests towards the effects of this environmental chemical on neurobiological functions of aquatic vertebrates of which little is known. In this study, the effects of bisphenol A on the distribution of the biologically more active somatostatin receptor subtypes in diencephalic regions of the teleost fish Coris julis were assessed using nonpeptide agonists (L-779, 976 and L-817, 818) that are highly selective for subtype(2) and subtype(5), respectively. Bisphenol A proved to be responsible for highly significant increased binding levels of subtype(2) in hypothalamic areas, while markedly decreased levels of subtype(5) were found in these diencephalic areas, as well as in the medial preglomerular nucleus. The extensive distribution of somatostatin receptor subtype(2) and subtype(5) in the teleost diencephalic areas suggests that, like in mammals, this receptor system may not only be involved in enhanced hypophysiotropic neurohormonal functions but might also promote neuroplasticity events.

Animals↗
PubMed2004

Preference, adaptation and survival of Mycoplasma pneumoniae subtypes in an animal model.

The interaction between Mycoplasma pneumoniae and its natural host, humans, cannot be studied directly for obvious reasons. Therefore, we used guinea pigs instead, which had been recently introduced as an acceptable alternative host organism. The following experimental approaches were taken to study the pathogen-host relationship: characterization and subtyping of M. pneumoniae strains isolated from human patients, infection of guinea pigs with selected M. pneumoniae strains, and analysis of adaptation, preference and survival of individual strains in guinea pigs under competitive conditions. The results of our study indicated that the species M. pneumoniae is genetically very homogeneous. From 115 independently isolated strains two subtypes and one variant were found. The subtypes differed significantly in the amino acid composition of the P1 protein, the main adhesin of M. pneumoniae, while the variant showed only minor amino acid exchanges. Infection of guinea pigs indicated differences between the subtypes and the variant in their ability to colonize and survive in the animal. Preinfection of the host with a certain subtype or variant caused a subtype-specific immunity and had a strong influence on the type of surviving bacteria in superinfection experiments. The results of these studies explain the shift of subtypes frequently observed in epidemic outbreaks of M. pneumoniae infection appearing in intervals of 3-7 years.

Adaptation, Physiological↗
PubMed2005

Single-channel properties of N- and L-subtypes of acetylcholine receptor in Ascaris suum.

We are interested in the properties of the target site of cholinergic anti-nematodal drugs for therapeutic reasons. The target receptors are ligand-gated ion channels that have different subtypes, and each subtype may have a different pharmacology. In a contraction assay using the parasitic nematode Ascaris suum, our laboratory has identified several subtypes, including an N-subtype, preferentially activated by nicotine, and an L-subtype, preferentially activated by levamisole. Here we use patch-clamp recordings to test the hypothesis that the single-channel selectivities of nicotine and levamisole are different. Unitary currents evoked by nicotine in this preparation were characterised for the first time. In some patches, both nicotine and levamisole activated small- and large-conductance channels. In other patches, the agonists activated just one channel amplitude. Discriminant analysis allowed classification of the one-conductance patch channels into the small or large categories, based on sets defined by the two-conductance patch data. The small channels had a conductance of 26.1+/-1.5 pS, n=18 (mean+/-SEM); the large conductance channels had a conductance of 38.8+/-1.2 pS, n=23 (mean+/-SEM). Analysis of amplitude histograms of the two-conductance patches showed that nicotine preferentially activated the small-conductance channels and levamisole preferentially activated the large-conductance channels. Our observations suggest that the N-subtype receptor channel has a conductance of 26 pS channel and the L-subtype receptor channel has a conductance of 39 pS.

Animals↗
PubMed2004

Genetic subtyping of influenza A viruses using RT-PCR with a single set of primers based on conserved sequences within the HA2 coding region.

Influenza A viruses are subtyped conventionally according to the antigenic characteristics of the external glycoproteins, haemagglutinin (HA) and neuraminidase (NA). To date 15 HA and 9 NA subtypes have been described. There is a need to develop fast, accurate and reliable methods to identify influenza virus subtypes, which may be associated with disease outbreaks. An RT-PCR is described using a single primer pair based on a conserved region of the HA2 gene that can detect all 15 HA influenza A subtypes. The assay was validated initially using a panel of 12 known standard prototype strains of influenza virus representing 6 HA subtypes and subsequently in a blind study using a panel of 30 strains. Selected viruses represented all known HA subtypes derived from avian, swine and human hosts separated both geographically and with time Sequence analysis of RT-PCR product showed complete correlation with results obtained using conventional serological methods. It is concluded that this RT-PCR is a reliable, robust and reproducible tool for the rapid identification of a wide range of all the HA subtypes of influenza A viruses.

DNA Primers↗
PubMed2005

Optimization of a genotypic assay applicable to all human immunodeficiency virus type 1 protease and reverse transcriptase subtypes.

Genotypic assays are used often to guide clinicians in decisions concerning the treatment of patients. An optimized sequence-based genotypic assay was used to determine the whole protease and reverse transcriptase (RT) gene, including the gag cleavage site region and RNase H region. Since non-B subtypes are increasing in countries where subtype B was the most prevalent subtype, and treatment becomes more available in developing countries where the epidemic is characterized by a high prevalence of non-B subtypes, it was important that the genotypic test was evaluated using a panel of different subtypes. Amplification was successful for different subtypes: A, B, C, D, F, G, H, J, CRF01_AE, CRF02_AG, CRF11_cpx, CRF13_cpx and an uncharacterized recombinant sample. The detection limit of the PCR was 1000 copies/ml, except for 1 subtype C sample (PL3) and 1 CRF02_AG sample (PL8). The detection limit for these samples was 5000 copies/ml. A sequence could be obtained in both directions for most of the samples.

DNA Primers↗
PubMed2004

Alpha1 adrenoceptor subtypes in human urinary bladder: sex and regional comparison.

A detailed study of the presence of alpha1 AR binding sites and alpha1 AR subtype mRNA expression in human urinary bladder areas involved in the micturition (i.e. detrusor, trigone and neck) is reported here, investigating whether or not there are differences between sexes. Results obtained indicated that alpha1 AR proteins were detectable in each bladder area. In both sexes, the detrusor and the neck expressed similar levels of alpha1 ARs: respectively, detrusor: 14.6 +/- 1.2 in men and 13.1 +/- 1.1 fmol/mg prot in women; neck: 16.9 +/- 3.2 in men and 17.5 +/- 4.1 fmol/mg prot in women. In the trigone, significantly higher alpha1ARs were found in women compared to men (20.6 +/- 1.1 vs 11.7 +/- 0.7 fmol/mg prot). Subtype analysis indicated that in women, each area was endowed with mRNA encoding for each alpha1 AR subtype. The men detrusor expressed alpha1a and alpha1d ARs, while in the trigone and the neck, each subtype was present. Since the detrusor muscle hypertrophy is a marker of bladder obstructive outlet, the selective alpha1 AR subtype targeting arouses much interest, as evidence indicates that there are differences in signalling pathways among the subtypes. Furthermore, the significance of the alpha1 ARs coexpression is still unknown; interestingly, recent papers demonstrate that alpha1 AR subtypes could dimerize. Thus, in the human urinary bladder it may be suggested a potential level of alpha1 AR complexity that could have an impact on drug development.

Aged↗