A random parameters with heterogeneity in means and Lindley approach to analyze crash data with excessive zeros: A case study of head-on heavy vehicle crashes in Queensland.

This study performed statistical analyses to identify likely crash contributing factors for Head-on Fatal and Serious Injury (FSI) collisions involving heavy vehicles (HVs) on the Queensland state road network. Head-on HV collisions are associated with the largest number of fatalities compared to other crash types in Queensland. However, there is limited relevant literature regarding this type of crashes. Recent studies on road safety research have focused on variants of random parameters models to capture unobserved heterogeneity that may influence the occurrence of crashes. Among such models, random parameters with heterogeneity in means has recently provided better results and has become popular. However, this study illustrates a potential limitation regarding the use of these models without explicitly factoring for excessive zero crash observations. To address this potential limitation, a random parameters with heterogeneity in means and a Lindley distribution is introduced in this study to factor for the unobserved heterogeneity using additional variables as well as site-specific variation from excessive zero crash observations. Results showed that a Poisson model with random parameters and heterogeneity in means using a Lindley distribution outperformed multiple alternative state-of-the-art specifications in terms of fit as well as overall prediction ability. The analyses using the proposed modelling approach revealed factors likely to affect the likelihood of Head-on FSI crashes involving HVs in Queensland including volume, segment length, period of analysis, terrain type being rolling, curve (moderate/sharp/very sharp) longer than 50% of the corresponding segment length, rural single carriageway with high (>=100 kph) and medium (>=50 and <100 kph) speed limits, and urban single carriageway. Unobserved heterogeneity regarding the parameter for road curvature was explained using rolling terrain type as an explanatory variable. This study has explained variation in the means of random parameters for a road attribute using the effect of a geometric variable, in which several stakeholders are primarily interested.

Molecular dynamics of amorphous pharmaceutical fenofibrate studied by broadband dielectric spectroscopy$ / 药物分析学报

Fenofibrate is mainly used to reduce cholesterol level in patients at risk of cardiovascular disease. Thermal transition study with the help of differential scanning calorimetry (DSC) shows that the aforesaid active pharmaceutical ingredient (API) is a good glass former. Based on our DSC study, the molecular dynamics of this API has been carried out by broadband dielectric spectroscopy (BDS) covering wide temperature and frequency ranges. Dielectric measurements of amorphous fenofibrate were per-formed after its vitrification by fast cooling from a few degrees above the melting point (Tm ? 354.11 K) to deep glassy state. The sample does not show any crystallization tendency during cooling and reaches the glassy state. The temperature dependence of the structural relaxation has been fitted by single Vogel–Fulcher–Tamman (VFT) equation. From VFT fit, glass transition temperature (Tg) was estimated as 250.56 K and fragility (m) was determined as 94.02. This drug is classified as a fragile glass former. Deviations of experimental data from Kohlrausch–Williams–Watts (KWW) fits on high-frequency flank of α-peak indicate the presence of an excess wing in fenofibrate. Based on Ngai's coupling model, we identified the excess wing as true Johari–Goldstein (JG) process. Below the glass transition temperature one can clearly see a secondary relaxation (γ) with an activation energy of 32.67 kJ/mol.

AE9C90CB: a novel, bladder-selective muscarinic receptor antagonist for the treatment of overactive bladder.

BACKGROUND AND PURPOSE: AE9C90CB (N- [(1R, 5S, 6R)-3-azabicyclo [3.1.0] hex-6-ylmethyl]-2-hydroxy-N-methyl-2, 2-diphenylacetamide), a novel muscarinic receptor antagonist, was synthesized for the treatment of overactive bladder. Here we describe the in vitro and in vivo profiles of AE9C90CB for action in bladder over salivary gland and compare it with four agents already in clinical use (tolterodine, oxybutynin, solifenacin and darifenacin). EXPERIMENTAL APPROACH: Radioligand binding assay and isolated tissue-based functional assay were used to evaluate affinity, potency, and receptor subtype selectivity of compounds. Inhibition of carbachol-induced increase in intravesicular pressure and salivary secretion were measured in anaesthetized rabbits to assess the functional selectivity. KEY RESULTS: In vitro radioligand binding study using human recombinant muscarinic receptors showed that AE9C90CB had greater affinity for M(3) muscarinic receptors with pKi of 9.90 +/- 0.11 and was 20-fold more selective for M(3) than for M(2) muscarinic receptors. AE9C90CB exhibited an unsurmountable antagonism on rat bladder strips (pK(B), 9.13 +/- 0.12). In anaesthetized rabbits after intravenous administration, AE9C90CB dose dependently inhibited carbachol-induced increase in intravesicular pressure and salivary secretion, and exhibited functional selectivity for urinary bladder over salivary gland which was ninefold better than that of oxybutynin. CONCLUSIONS AND IMPLICATIONS: We have identified AE9C90CB, a compound exhibiting moderate selectivity for M(3) over M(2) receptors but greater selectivity for urinary bladder over salivary gland than oxybutynin, tolterodine, solifenacin and darifenacin. Therefore, AE9C90CB may be a promising compound for the treatment of overactive bladder with reduced potential to cause dry mouth than currently available antimuscarinic drugs.

Benign prostatic hyperplasia: an insight into current investigational medical therapies.

Benign prostatic hyperplasia (BPH) is a leading disorder of the elderly male population that is characterised by a progressive enlargement of prostatic tissue, resulting in obstruction of the proximal urethra and causing urinary flow disturbances. The pathophysiology of BPH associated with lower urinary tract symptoms is characterised by increased adrenergic tone (dynamic component) leading to smooth muscle contraction and prostatic overgrowth due to androgenic stimulation (static component); therefore, the therapeutic armamentarium of BPH can be broadly divided into antiadrenergic and antiandrogenic approaches. alpha1-Adrenoceptor antagonists and 5alpha-reductase inhibitors are well-established representatives of the two categories, respectively. Other antiandrogenic approaches involve gonadotropin-releasing hormone agonists and antagonists for the treatment of prostate hyperplasia. Apart from these approaches, new approaches with novel targets are emerging. The advent of new therapies is, however, more oriented towards the static component. These involve metabolic factors (hexokinase inhibitor), growth factors (vitamin D3 analogues), oxytocin antagonists and gonadotropin-releasing hormone Gi agonist-based therapies. Gene therapy and photodynamic therapies are other emerging therapies for relieving symptoms in BPH patients. With the initial success of upcoming targets, the unmet need to develop an efficacious and relatively safe therapeutic modality is discussed. Nevertheless, their long-term safety and efficacy needs to be evaluated in large-scale clinical trials. The future also belongs to combination therapies to combat both dynamic and static disease components and for extended indications such as micturition disorder and non-bacterial prostatitis.

Raf-1 expression may influence progression to androgen insensitive prostate cancer.

BACKGROUND: Recent evidence has implicated the MAP kinase pathway with the development of androgen insensitive prostate cancer (AIPC). We have previously reported gene amplification of critical members of this pathway with the development of androgen insensitive disease. METHODS: Protein expression of Raf-1 was analyzed using immunohistochemistry (IHC) in a database of 65 paired tumor specimens obtained before and after the development of AIPC and correlated with other members of the pathway. RESULTS: Patients whose Raf-1 expression rose with development of AIPC had a significantly shorter median time to biochemical relapse compared to those whose expression fell or remained unchanged (1.16 vs. 2.62 years, P = 0.0005). In AIPC tumors, expression of Raf-1 correlated significantly with expression of HER2 and with expression of c-fos. CONCLUSIONS: We conclude that the HER2/Raf-1/AP-1 axis may promote the development of AIPC, leading to early relapse. Members of the pathway may act as novel therapeutic targets for patients.

Androgen receptor gene amplification and protein expression in hormone refractory prostate cancer.

This study examined androgen receptor (AR) gene amplification and protein expression in 102 matched paired hormone sensitive and resistant tumours from 51 patients. AR gene amplification and X chromosome copy number were assessed by fluorescent in situ hybridisation, and protein expression was assessed by immunohistochemistry. All tumours were stained for PSA protein expression. Significantly more tumours exhibited AR amplification following the development of hormone resistance (20%, 10 out of 49) compared to matched hormone-sensitive tumours from the same patient (2%, one out of 48) (P=0.0085). The level of AR expression was significantly higher in hormone-resistant tumours compared to matched hormone-sensitive tumours from the same patient (130, interquartile range, 55-167 vs 94.5 interquartile range, 55-120, P=0.019). AR expression levels in hormone-resistant tumours with and without AR amplification were not significantly different. However, an increase in AR expression was seen with the development of AR amplification in paired tumours. The rate of AR gene amplification and/or an increase in AR protein expression during androgen resistant is too low to wholly explain the development of androgen resistance. Alternative mechanisms for modulating the function of the AR, or other signalling pathways, must be considered as key factors in the development of hormone-resistant prostate.

Amplification of the androgen receptor may not explain the development of androgen-independent prostate cancer.

OBJECTIVE: To examine the role of androgen receptor (AR) gene amplification and aneusomy of the X chromosome in the development of antiandrogen-resistant prostate cancer. PATIENTS AND METHODS: Twenty patients with prostate cancer resistant to androgen-deprivation therapy were selected for study. The records of patients with tumours before and after antiandrogen therapy, and with a full clinical follow-up, were retrieved. AR gene amplification and X chromosome copy number were assessed by fluorescence in situ hybridization using a labelled probe at locus Xq11-13 for the AR gene and a labelled alpha-satellite probe for the X chromosome. At least 20 nuclei were scored over three tumour areas by two independent observers. RESULTS: Aneusomy of the X chromosome was reported respectively in seven (35%) and 11 (55%) tumours before and after hormone relapse, the AR gene copy number was increased in seven (35%) and 13 (65%), respectively, and AR gene amplification was detected in one (5%) and three (15%), respectively. Neither increased AR copy number nor AR amplification in primary tumours precluded a biological response to androgen-deprivation therapy. CONCLUSION: The rate of AR gene amplification is too low to be solely responsible for the development of antiandrogen-resistant prostate cancer. Also, the presence of amplified AR and cells aneusomic for the X chromosome in primary tumours that respond to androgen-deprivation therapy suggests that an increase in AR gene copy number does not prevent a tumour from responding to this therapy. Therefore other mechanisms which could cause hormone-refractory prostate cancer must be investigated before it is understood why so many patients relapse with this disease.

Epidermal growth factor receptor mRNA and protein are expressed in progenitor cells of the olfactory epithelium.

Olfactory receptor neurons are continuously replaced postnatally through the initiation of the division and terminal differentiation of progenitor cells located in the basal layer of the olfactory epithelium. Although the factors that regulate this process in vivo are not known, recent in vitro studies demonstrated that members of the epidermal growth factor (EGF) family including transforming growth factor-alpha (TGF alpha) and EGF are highly potent in promoting the proliferation of progenitor cells, suggesting a role for the EGF receptor (EGFR), which is the molecular receptor for both mitogens. We have examined the expression of EGFR mRNA and protein in the olfactory epithelium by using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis and have examined their cellular localization with in situ RT-PCR and immunocytochemistry. RT-PCR and Southern blot analysis demonstrated that EGFR mRNA is expressed in the olfactory mucosa and also in the positive control tissues, kidney and tongue. The 170-kDa EGFR protein was identified with Western blot analysis in the olfactory epithelium and control tissues. Our results using in situ RT-PCR localized EGFR mRNA-expressing cells more extensively in the basal cell layer of the epithelium than did the immunocytochemical methods. These results suggest that EGFR mediates the mitogenic effect of TGF alpha and/or EGF on the quiescent basal cells to initiate the cell cycle.

Activation of hypothalamic neurons by intraovarian pressure signals in a teleost fish, Clarias batrachus: role of mechanosensitive channels.

Application of intraovarian pressure is known to trigger profound cytomorphological changes in the neurosecretory cells of nucleus preopticus in the teleost Clarias batrachus. These findings indicate the presence of stretch receptors in the ovaries, perhaps equipped with mechanosensitive channels that transduce the stretch signals to be transmitted to the brain. To test the occurrence of the mechanosensitive channels in the ovaries, we administered a range of pharmacological agents (lignocaine, quinidine, tetraethylammonium chloride, ethylene-diaminetetraacetic acid and gadolinium) known to block the mechanosensitive ion channels, in the ovarian lumen prior to the administration of the intraovarian pressure. Pretreatment with the above agents inhibited the response by the nucleus preopticus neurosecretory cells to intraovarian pressure. The results suggest the occurrence of the mechanosensitive channels in the ovaries of teleostean fishes. In terms of function we speculate that the stretch sensory system and the ensuing pathway connecting the ovaries with the hypothalamus might play a role in apprising the brain of the status of ovarian maturity and in the initiation of the spawning reflex.

Age- and gender-related trends in the expression of glutathione S-transferases in human nasal mucosa.

The cellular expression of alpha, mu, and pi classes of glutathione S-transferases (GSTs) was investigated in human nasal mucosa by means of immunocytochemical techniques. In the olfactory mucosa, immunoreactivity for GST-alpha was most intense in the acinar cells of the Bowman's glands, with weak immunoreactivity in the supranuclear region of sustentacular cells. Whereas GST-pi was localized only in the sustentacular cells, no GST-mu was detected. In the respiratory mucosa, GST-alpha and GST-pi were detected at the brush borders of ciliated columnar epithelial cells. There were age- and gender-related trends in the expression of GST-alpha, but not GST-pi, in the olfactory mucosa. The intensity of immunoreactivity in the olfactory mucosa was decreased in older subjects. The expression of GST-alpha in the olfactory mucosa of females consistently exhibited greater intensity than that of males at all the ages studied. These differences were not observed in the respiratory mucosa. These results indicate that acinar cells of the Bowman's glands and sustentacular cells are the major sites of phase II biotransformation in the human nasal mucosa.

Human olfactory receptor neurons contain OMP mRNA in their dendritic and axonal processes.

The cellular expression of olfactory marker protein (OMP) mRNA and protein was investigated in the olfactory mucosa of humans ranging in age from 26 weeks of gestation to 85 years using in situ hybridization and immunocytochemistry. OMP mRNA and protein were most abundant in the somas of olfactory receptor neurons (ORNs). The hybridization signal over the ORN somal layer was greater in older subjects than in younger ones, reflecting either a higher neuronal density or more OMP mRNA per cell. In contrast, it was significantly lower in subjects with Alzheimer's disease when compared with an age-matched control. Characteristics of older subjects were patchiness in the distribution of OMP-expressing ORNs and the occurrence of subepithelial invaginations containing OMP-positive neurons. In addition, a significant hybridization signal was detected in the apical olfactory epithelium containing the dendrites, dendritic knobs, and cilia of ORNS, and over olfactory nerve bundles in the lamina propria, indicating the occurrence of OMP mRNA in dendritic and axonal domains.

Human olfactory receptor neurons express heat shock protein 70: age-related trends.

Immunocytochemical methods were used to investigate the cellular distribution and age-related trends in the expression of constitutive and/or inducible forms of heat shock protein (hsp) 70 in the human nasal mucosa of 22 subjects who ranged in age from 16 weeks prenatal to 90 years, including 3 subjects with Alzheimer's disease. The olfactory mucosa was characterized by the presence of olfactory marker protein-immunoreactive olfactory receptor neurons. The hsp 70 immunoreactivity was localized in olfactory receptor neurons and the supranuclear region of sustentacular cells in the olfactory epithelium, and in the acinar cells of the Bowman's glands in the lamina propria. A systematic age-related decrement in the expression of hsp 70 immunoreactivity was observed in the olfactory receptor neurons. This trend was not apparent in sustentacular cells and Bowman's glands. A marked decrement in hsp 70 immunoreactivity was also noted in the olfactory receptor neurons of subjects with Alzheimer's disease when compared to age-matched controls. These results suggest that the age-dependent decrease in hsp 70 in olfactory receptor neurons of older subjects and those with Alzheimer's disease may be attributable to their greater susceptibility to stress.

Differential expression of vomeromodulin and odorant-binding protein, putative pheromone and odorant transporters, in the developing rat nasal chemosensory mucosae.

Expression of the putative pheromone and odorant transporter, vomeromodulin, was characterized in developing rat nasal mucosae using in situ hybridization and immunocytochemistry. Initial expression of vomeromodulin mRNA and protein was detected at embryonic day (E)16 in the maxillary sinus component of the lateral nasal glands. The abundance of mRNA and protein in the lateral nasal glands increased with age and reached a peak at postnatal day (P)27. Also at P27, vomeromodulin mRNA and protein expression was initiated in vomeronasal glands and posterior glands of the nasal septum. Comparison of the developmental expression of odorant-binding protein, another carrier protein synthesized in the lateral nasal glands, with that of vomeromodulin demonstrated major differences. In contrast to vomeromodulin, odorant-binding protein was not detected until postnatal day 2 in the ventral component of the lateral nasal glands and anterior glands of the nasal septum. These results suggest that the expression of vomeromodulin and odorant-binding protein is developmentally and differentially regulated and confirms the suggestion that vomeromodulin may function in olfactory and vomeronasal perireceptor processes as a transporter for pheromones and odorants. In addition, the embryonic expression of vomeromodulin suggests its involvement in olfactory perireceptor processes in utero.

Expression of the putative pheromone and odorant transporter vomeromodulin mRNA and protein in nasal chemosensory mucosae.

In nasal chemosensory systems, glandular proteins associated with the vomeronasal and olfactory epithelia perform specific perireceptor functions associated with sensory transduction. Vomeromodulin, a recently identified glycoprotein synthesized by the lateral nasal glands, is proposed to be a pheromone transporter (Khew-Goodall et al., FASEB J 5:2976-2982, 1991). In our study, we have investigated its expression in vomeronasal, olfactory, and respiratory nasal mucosae of rats and humans using in situ hybridization and immunocytochemical techniques. In the rat, vomeromodulin mRNA and protein were localized abundantly in the glandular acini of the maxillary sinus component of the lateral nasal glands. In addition, the vomeronasal and posterior glands of the nasal septum also expressed vomeromodulin mRNA and protein. Vomeromodulin immunoreactivity was localized extracellularly in the mucus of the sensory and non-sensory epithelia of the vomeronasal organ, and in the mucociliary complex of the olfactory, respiratory, and associated nasal epithelia. In human nasal mucosae, vomeromodulin immunoreactivity was localized in the mucociliary complex of the vomeronasal and respiratory epithelia. Comparison of the localization of vomeromodulin with that of odorant-binding protein, which is also synthesized in the lateral nasal glands of rats, revealed that odorant-binding protein was expressed in a completely separate glandular region, namely the ventral component. In the septal glands, vomeromodulin was expressed in the posterior glands whereas odorant-binding protein was localized in the anterior glands. Odorant-binding protein immunoreactivity was not observed in the vomeronasal glands. In contrast, both proteins were localized in the mucus of vomeronasal, olfactory, and respiratory epithelia. Our results suggest that vomeromodulin, like odorant-binding protein, functions as a chemosensory stimulus transporter associated with perireceptor processes in vomeronasal and olfactory transduction.