Position paper on olfactory dysfunction.

BACKGROUND: Olfactory dysfunction is an increasingly recognised condition, associated with reduced quality of life and major health outcomes such as neurodegeneration and death. However, translational research in this field is limited by heterogeneity in methodological approach, including definitions of impairment, improvement and appropriate assessment techniques. Accordingly, effective treatments for smell loss are limited. In an effort to encourage high quality and comparable work in this field, among others, we propose the following ideas and recommendations. Whilst the full set of recommendations are outlined in the main document, points include the following: - Patients with suspected olfactory loss should undergo a full examination of the head and neck, including rigid nasal endoscopy with small diameter endoscopes. - Subjective olfactory assessment should not be undertaken in isolation, given its poor reliability. - Psychophysical assessment tools used in clinical and research settings should include reliable and validated tests of odour threshold, and/or one of odour identification or discrimination. - Comprehensive chemosensory assessment should include gustatory screening. - Smell training can be helpful in patients with olfactory loss of several aetiologies. CONCLUSIONS: We hope the current manuscript will encourage clinicians and researchers to adopt a common language, and in so doing, increase the methodological quality, consistency and generalisability of work in this field.

Pulmonary delivery of peptide YY for food intake suppression and reduced body weight gain in rats.

AIMS: Peptide YY (PYY) is an endogenous anorectic gut-secreted peptide that has been shown to suppress appetite in animals and humans, when given by injection. This study tested if needle-free pulmonary delivery of PYY enables food intake suppression and reduced body weight gain in rats. The PYY pharmacokinetics and effects on brain neuropeptide levels were also examined. METHODS: Rats received single or once-daily 7-day pulmonary administration of saline or PYYs. Food intake and body weight gain were monitored to study the effects of different doses (0.08-0.90 mg/kg) of PYY3-36, PYY1-36 and PYY13-36. Plasma PYY pharmacokinetics were determined via enzyme-linked immunosorbent assay. Changes in orexigenic neuropeptide Y (NPY) and c-Fos protein levels in the hypothalamus arcuate nucleus (ARC) were measured by immunofluorescence microscopy. RESULTS: PYY3-36 caused dose-dependent and 4- to 6-h food intake suppression following pulmonary delivery. At 0.80 mg/kg, the effect was significant with 35.1 ± 5.7 and 19.7 ± 4.2% suppression at 4 and 6 h, respectively. Repeated administration for 7 days reduced cumulative body weight gain by 39.4 ± 11.0%. PYY1-36, but not PYY13-36, was equipotent to PYY3-36 in food intake suppression. The plasma PYY concentration reached its peak at 10 min following pulmonary delivery with 12-14% of bioavailability. Increased c-Fos and reduced NPY expressions were observed in the hypothalamus ARC, consistent with the magnitude of food intake suppression by each of the PYYs. CONCLUSIONS: Pulmonary delivery of PYY enabled significant 4- to 6-h food intake suppression via 12-14% of lung absorption and hypothalamic ARC interaction, leading to reduced body weight gain in rats.

Multiple effectiveness aspects of tapentadol for moderate-severe cancer-pain treatment: an observational prospective study.

BACKGROUND: Previous studies have shown the efficacy of tapentadol (TP) for chronic cancer pain. We evaluated multiple effectiveness aspects of TP prolonged release on moderate-severe cancer-related pain, neuropathic pain (NeP), patient satisfaction, and quality of life. METHODS: An observational prospective study was conducted on 80 cancer patients. Opioid-naïve patients received a starting dose of prolonged-release TP 50 mg twice daily, and opioid-experienced patients were switched to TP, not to exceed 500 mg/day. Treatment response was evaluated at 3, 6, 30-40, and 60-70 days through response rate, numeric rating-scale scoring, survival analysis (time to event for response), pain-intensity difference, TP escalation-index percentage, and effects on NeP. The drug-sparing effect on concomitant therapies was evaluated. RESULTS: Seventy of 80 patients (88%) were responders to treatment (95% CI 78%-94%). Compared to T0, pain-intensity reductions were statistically significant for all intervals (P<0.01), with better results at T3/T4. NeP was significantly reduced at T4 (P<0.01). The probability of response was low at the initial stages and increased during the study. Pain-intensity differences decreased during the study, though without significance. Two patients (2.5%) left the study for TP-induced side effects. A significant improvement in quality of life was observed after 30-40 days (P<0.01). The majority of patients were "satisfied", "very satisfied", or "extremely satisfied" (T3-T4). CONCLUSION: TP was effective in terms of drug-sparing effect, response rate, TP escalation-index percentage, and NeP management. By comparing data from the survival analysis with the response rate and time to response (numeric rating scale from T0 to T4), we found that although TP induced a quick response, a longer period of therapy and higher doses were needed to improve the positive result.

Contribution of Helicobacter pylori infection to the risk of peptic ulcer bleeding in patients on nonsteroidal anti-inflammatory drugs, antiplatelet agents, anticoagulants, corticosteroids and selective serotonin reuptake inhibitors.

BACKGROUND: Nonsteroidal anti-inflammatory drugs, low-dose aspirin, non-aspirin antiplatelet agents, anticoagulants, selective serotonin reuptake inhibitors and corticosteroids increase the risk of gastroduodenal bleeding. AIM: To determine in a retrospective cohort study the contribution of Helicobacter pylori infection to the risk of peptic ulcer bleeding in patients taking these drugs. METHODS: Among patients with peptic ulcer disease diagnosed by endoscopy from 01/2004 to 12/2014 (N = 1719, 60% males, age 65.8 ± 14.5), 56.9% had peptic ulcer bleeding (cases) and 43.1% uncomplicated peptic ulcer disease (controls). Demographics, intake of nonsteroidal anti-inflammatory drugs, aspirin, non-aspirin antiplatelet agents, anticoagulants, selective serotonin reuptake inhibitors, proton pump inhibitors and corticosteroids were documented. H. pylori status was determined by histology, rapid urease test or serology. Adjusted odds ratios (OR) were estimated by logistic regression analysis. RESULTS: Helicobacter pylori infection increased the risk of peptic ulcer bleeding in nonsteroidal anti-inflammatory drug and aspirin users (OR = 2.91, 95% CI = 1.71-4.98 and OR = 2.23, 95% CI = 1.52-3.28, respectively), but not in patients on anticoagulants, selective serotonin reuptake inhibitor or corticosteroid therapy. H. pylori-positive status substantially increased the risk of peptic ulcer bleeding in patients on non-aspirin antiplatelet agents (OR = 4.37, 95% CI = 1.28-14.99), concomitant aspirin/nonsteroidal anti-inflammatory drug intake (OR = 5.85, 95% CI = 1.68-20.36) and combined antiplatelet therapy (OR = 8.43, 95% CI = 1.09-65.17). After further adjustment for proton pump inhibitor intake, H. pylori infection was still a risk factor for peptic ulcer bleeding in nonsteroidal anti-inflammatory drug and aspirin users. CONCLUSIONS: Helicobacter pylori infection increases the risk of peptic ulcer bleeding in peptic ulcer disease patients on nonsteroidal anti-inflammatory drugs, aspirin and non-aspirin antiplatelet agents. H. pylori-positive patients on combined antiplatelet therapy carry the highest risk for peptic ulcer bleeding.

Activity of Berberis aetnensis root extracts on Candida strains.

The antifungal activity of methanolic extract and alkaloidal fraction of Berberis aetnensis against Candida species was investigated. The crude extract was active against Candida species, this activity being higher than that of the alkaloidal fraction and berberine.

Bilateral hydrocele. Uncommon clinical presentation of primary testicular lymphoma in the elderly.

Primary testicular lymphoma (PLT) represents 5% of testis tumors, the incidence increases in patients older than 60 years of age. Bilateral hydrocele is an unusual clinical presentation. Relapse in the central nervous system and in the contralateral testis is often observed. The US shows hypoechoic nodular lesions with a complete structural involvement of didymus and hypervascularization at Color Doppler. Orchiectomy should be performed in all cases as it is indispensable for the histopathological diagnosis and to characterize the immunophenotypic features. The most common histotype is diffuse large-B cell lymphoma. Combined biological approach and chemotherapy with rituximab and doxorubicin has radically changed the prognosis of disease. The authors report two patients of 81 and 82 years-old who referred for evaluation of massive bilateral hydrocele causing severe limitation of deambulation. Negative cytological findings for neoplastic cells in the scrotal effusion made difficult the differential diagnosis between inflammatory and malignant disease. Histopathologic findings made a diagnosis of high grade diffuse large B-cell NHL, respectively stage IV-E and stage III-E. The 82 years old patient was treated with 6 chemotherapy cycles of rituximab, cyclophosphamide, vincristine, prednisone. The exitus was dued to the umbilical hernia complications. In the 81 years old patient, cognitive deficit and severe impairment of general conditions constituted an absolute contraindication to polychemotherapy treatment. Rapid tumor progression led the patient to exitus 2 months after diagnosis. In both patients the delayed diagnosis of PLT was probably due to the reduction of welfare protection in the elderly with adverse social conditions.

Position paper on olfactory dysfunction.

Background: Olfactory dysfunction is an increasingly recognised condition, associated with reduced quality of life and major health outcomes such as neurodegeneration and death. However, translational research in this field is limited by heterogeneity in methodological approach, including definitions of impairment, improvement and appropriate assessment techniques. Accordingly, effective treatments for smell loss are limited. In an effort to encourage high quality and comparable work in this field, among others, we propose the following ideas and recommendations. Whilst the full set of recommendations are outlined in the main document, points include the following: • Patients with suspected olfactory loss should undergo a full examination of the head and neck, including rigid nasal endoscopy with small diameter endoscopes. • Subjective olfactory assessment should not be undertaken in isolation, given its poor reliability. • Psychophysical assessment tools used in clinical and research settings should include reliable and validated tests of odour threshold, and/or one of odour identification or discrimination. • Comprehensive chemosensory assessment should include gustatory screening. • Smell training can be helpful in patients with olfactory loss of several aetiologies. Conclusions: We hope the current manuscript will encourage clinicians and researchers to adopt a common language, and in so doing, increase the methodological quality, consistency and generalisability of work in this field.

Antibacterial activity of propolis and its active principles alone and in combination with macrolides, beta-lactams and fluoroquinolones against microorganisms responsible for respiratory infections.

Propolis is produced by bees and is reported to have several pharmaceutical properties. Its antibacterial activity against strains causing upper respiratory tract infections is particularly important: propolis might be used as a therapeutic agent to prevent the bacterial infections that sometimes overlap viral infections. In this study the in vitro activity of both an alcoholic solution and a hydroglyceric extract of propolis, as well as its active principles, was tested against bacteria responsible for respiratory infections (Streptococcus pneumoniae, Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis and Streptococcus pyogenes). We also evaluated the in vitro activity of a combination of propolis and its active principles and some beta-lactams, macrolides and fluoroquinolones. Our results, though not demonstrating a clearly synergistic activity between antibiotics and propolis and its constituents, show the possibility of using natural preparations, due to their antimicrobial and anti-inflammatory properties, to enhance antibacterial therapy.

Electrophysiological evidence for a topographical projection of the nasal mucosa onto the olfactory bulb of the frog.

Three olfactory nerve branches respectively subserving either a medial, an intermediate, or a lateral region of the dorsal olfactory receptor sheet of the bullfrog Rana catesbeiana were electrically stimulated with bipolar platinum hook electrodes. Extracellular single unit responses from 93 second-order cells in different regions of the olfactory bulb were recorded with metal-filled glass micropipets. The excitatory responsiveness of each unit to the stimulation of each of the three nerve branches (response profile) was determined. Some units were sensitive to stimulation of each of the three nerve branches, thus suggesting a wide projection from the entire receptor sheet. On the other hand, other units were more selective. Of this latter group, units in the lateral bulb were excited by nerve branches subserving the more lateral regions of the receptor sheet; units in the medial bulb were excited by the nerve branches subserving the more medial regions of the receptor sheet. These data provide electrophysiological evidence for a topographical projection of the olfactory receptor sheet onto the olfactory bulb, and further suggest that the projections onto different bulbar cells vary in degree of localization.

Receptive fields of second-order neurons in the olfactory bulb of the hamster.

Electrical stimulation of nerve fibers emerging from different positions of the olfactory epithelium was used to determine the receptive fields for 52 olfactory bulb neurons in the hamster. The responses of olfactory bulb neurons were recorded extracellularly with metal-filled micropipettes. Suprathreshold stimuli (500 microA) were applied to each of eight standard epithelial positions spaced approximately 250 microns apart, and the position, or positions, which, when stimulated, produced a response in the bulb were taken as an index of the neuron's receptive field. The results indicate that most bulb neurons have very localized receptive fields limited to only one or two stimulating positions. Furthermore, there was a statistically significant correlation between the location of a neuron's receptive field in the olfactory epithelium and the recording depth of the neuron in the olfactory bulb (Spearman rank correlation coefficient, rs, 0.67, P < 0.001). These findings demonstrate that in the mammalian olfactory system there exists a topographical projection of input from localized regions in the epithelium onto the second-order neurons in the olfactory bulb.

A quantitative analysis of changes in the olfactory epithelium following bulbectomy in hamster.

Continuous replacement of sensory neurons is a normal process in the olfactory system of adult vertebrates. The capacity for replacement following experimentally induced degeneration of sensory neurons makes this system ideal for the study of the dynamics of neural populations. A quantitative analysis was made to determine the time course and degree of cell replacement in the olfactory epithelium following unilateral bulbectomy in adult hamsters. Histological measurements of number of cells and epithelial thickness were made for up to 194 days postoperatively. Results for each experimental animal were expressed as a percentage of the contralateral control side. There was an immediate degeneration of cells, the number decreasing to 39% by day 4. During days 4--15 new growth resulted in an increase in cell number, which was maintained at a level of 60--70% through day 194. Epithelial thickness decreased to 60--70% during the degeneration period, but there was no recovery during subsequent days 4--194. Analysis of epithelial cells by cell type (supporting, receptor, and basal cells) showed that changes in cell numbers were limited to the neural cell populations (receptor and basal). This study confirms that olfactory sensory neurons are capable of replacement following degeneration in spite of the absence of normal target tissue. However, the observed recovery does not reach control levels and the functional capacity of replacement neurons requires further study.

Morphology of the human olfactory epithelium.

The human olfactory epithelium has been previously studied with scanning electron microscopy; however, most studies have been limited to examining the epithelial surface. In an attempt to examine structures below the surface, we scanned epithelial fractures that occurred during tissue preparation. This made it possible to obtain unique three-dimensional images of cell profiles from the mucosal surface through the full depth of the epithelium. We examined supporting cells, olfactory neurons, basal cells, and a fourth cell type, the microvillar cell. Supporting cells had a microvillar surface and were in close contact with olfactory neurons and their processes. Olfactory neurons were primarily located in the middle and lower epithelial regions. Basal cells occurred alone or in clusters adjacent to the basal lamina. Microvillar cells were always observed in the upper epithelial region. They were flask- or pear-shaped, had a tuft of microvilli that extended into the nasal cavity, and a thin axon-like process that passed basally towards the lamina propria. This study represents the first comprehensive scanning electron microscopy examination of the human olfactory epithelium. Three-dimensional images obtained for each epithelial cell type allowed us to examine cell processes and their close contacts, especially between supporting cells and olfactory neurons. These results also revealed the irregular and patchy distribution of olfactory receptors within the human nasal cavity. Further studies that examine the detailed morphology of the human olfactory epithelium should provide a better understanding of the physiological mechanism and clinical disorders that affect olfactory function in humans.

Morphology of olfactory epithelium in humans and other vertebrates.

Human olfactory epithelium is similar in organization and cell morphology to that of most vertebrate species. The epithelium has a pseudostratified columnar organization and consists of olfactory neurons, supporting and basal cells. Near the mucosal surface there are also microvillar cells. These cells have neuron-like features and may be chemoreceptors. Human olfactory epithelium is not a uniform sensory sheet. Patches of non-sensory tissue often appear in what was thought to be a purely olfactory region. The significance of these patches has not been determined, but they could reflect exposure to environment agents or changes that occur during the normal aging process. In order to better understand the human olfactory system, further knowledge of the normal structure is necessary. This review addresses the morphology of the human olfactory epithelium and the remarkable plasticity of the vertebrate olfactory system.

Comparison of neurogenesis and cell replacement in the hamster olfactory system with and without a target (olfactory bulb).

The olfactory sensory neurons are unique in the vertebrate nervous system in that they are replaced following experimentally induced degeneration. Unilateral removal of the olfactory bulb in hamster results in degeneration of all mature receptor neurons followed by a neurogenesis and partial replacement of the receptor cell population. To determine if full recovery requires the presence of normal target tissue, a study of sensory neuron replacement was made following a nerve transection procedure, which leaves the olfactory bulb (target) intact. A comparison of quantitative measurements of cell number and thickness in the sensory epithelium showed that the presence of the target tissue alone did not result in improved recovery. One possible explanation is that complete recovery requires that axons of newly formed receptors must first re-establish synaptic contact with the olfactory bulb. To test this possibility, it will be necessary to include longer postoperative recovery times than those used in the present study.

Neural regeneration and functional reconnection following olfactory nerve transection in hamster.

The olfactory sensory neurons in the vertebrate nervous system are unique in that they undergo continuous neurogenesis and replacement. Anatomical studies have shown that transection of the olfactory nerves leads to a degeneration of sensory neurons followed by a neurogenesis and replacement with newly formed cells. Replacement neurons grow axonal processes that are capable of reestablishing morphological connections with cells in the olfactory bulb. To determine the functional capacity of these anatomical reconnections, single unit responses to odor stimuli were recorded from cells in the olfactory bulb following recovery from unilateral olfactory nerve transection. A total of 56 cells were studied, taken from hamsters with recovery times of 4,35,60,90,120,180 and 270 days. At day 4, although there was spontaneous activity recorded from cells on the experimental side (n = 10), they did not respond to stimulation of the olfactory epithelium with odors. Control cells (n = 9) from the unoperated side of the same animals showed normal odor responses. By day 35, some of the cells tested on the experimental side responded to odor stimulation, indicating that connections had been reestablished with sensory neurons. With longer recovery times, an increasing percentage of cells responded to odor stimuli. In addition, concentration response functions showed that cells were capable of signaling differences in stimulus intensity. The response of cells to four odors (amyl acetate, 1-butanol, ethyl acetate and ethyl butyrate) showed differences in odor selectivity, suggesting their ability to discriminate among odors.(ABSTRACT TRUNCATED AT 250 WORDS)

Electrophysiological characterization of the olfactory bulb during recovery from sensory deafferentation.

Axonal transection results in degeneration of the olfactory sensory neurons (OSNs). Replacement OSNs reinnervate the olfactory bulb. To document reinnervation, lateral OSN fibers were stimulated while bulb negative evoked potentials (NEPs) were recorded. For control adult hamsters and at 20, 30, and 120 days after transection, lateral fibers were connected to the lateral more than the medial bulb. The percentage of lateral bulb positions with NEPs was similar to control at 30 and 120 days, but NEP amplitude did not reach control level.

Properties of kinesthetic neurons in somatosensory cortex of awake monkeys.

(1) To study neural mechanisms used to encode kinesthetic information in somatosensory cortex of awake monkeys, we recorded from 227 single neurons responsive to joint movement or specific postures of the forelimb or hand (kinesthetic neurons). Unit responses were characterized quantitatively with respect to: (a) firing patterns; (b) responses to ramp changes in joint position and joint velocity; and (c) responses to sinusoidal joint movements. (2) Kinesthetic neurons were divided into 3 groups. Rapidly-adapting neurons (44%) responded only to joint movement, giving a burst of impulses proportional to velocity. They showed no tonic responses to limb posture. Two populations of tonically active neurons were observed: slowly-adapting neurons (43%) and postural neurons (13%). Both types increased their firing rates with increasing degrees of flexion or extension, showing maximum excitation at the extremes of joint position in the preferred direction. They were distinguished by their sensitivity to the velocity of movement, the size of the angle over which they respond, and the phase relation of their responses to sinusoidal joint movement. (3) The firing rates of kinesthetic neurons in S-I cortex are functions of both joint angle and joint velocity. The importance of each component varies in the 3 classes: velocity of movement is the most important determinant of firing rates of rapidly-adapting and slowly-adapting kinesthetic neurons, and joint angle predominates the responses of postural neurons.

Multiple-joint neurons in somatosensory cortex of awake monkeys.

(1) In the somatosensory cortex of alert monkeys, 55 neurons were found which receive convergent information from two or more adjacent joints. Most of these multiple-joint neurons were excited by postures of the hand, particularly those involved in grasping. (2) Three basic types of joint interactions were observed. The simplest neurons (occlusion neurons) responded to postures of several different joints, but combination of the preferred postures produced no further increase in firing. The more complex cells showed summated responses to combined postures of adjacent joints, or subliminal facilitation between joints. The responses of both summation neurons and subliminal facilitation neurons were graded with joint angle, and there was an optimum or preferred position for both joints which gave the strongest response. (3) Multiple-joint neurons may provide a neuronal substrate for extracting postural information from several different populations of kinesthetic neurons. They therefore act as feature-detecting neurons, abstracting information about specific body postures.

Is nestin a marker for chemosensory precursor cells?

The vomeronasal and olfactory systems are unique in that their chemosensory neurons undergo continuous neurogenesis after development. Immunoreactivity to nestin, a neuronal precursor marker protein, was investigated in the developing rat vomeronasal organ to determine its potential as a cell marker. From postnatal day 1 (P1) to P22, the distribution of nestin positive cells became progressively restricted to the area adjacent to the basement membrane. By P29, the vomeronasal organ reached structural maturity and only a few nestin positive cells were observed. Results suggest that nestin may be a useful marker for neuronal precursor cells in studies of neurogenesis and development of chemosensory systems.

Regeneration of olfactory sensory neurons and reconnection in the aging hamster central nervous system.

Olfactory neuron recovery and axon growth was studied in 12-24 month old hamsters after unilateral olfactory nerve transection. At recovery times ranging from 4 to 126 days olfactory nerve regeneration and axon reconnection to the olfactory bulb were examined by anterograde horseradish peroxidase (HRP) neurohistochemistry and electron microscopy. Nerve transection produced immediate retrograde neuron degeneration and there was no HRP reaction product in the bulb at 4 days post transection. By day 35, centrally growing olfactory axons had reached the bulb. Axons formed glomeruli smaller than those in the unoperated control bulb and they were not always confined to the glomerular layer of the bulb. Some animals showed robust fiber growth with axon fascicles penetrating the different layers of the bulb and forming ectopic glomeruli along their path. Second order bulb neurons contained wheat germ agglutinin HRP reaction product, indicating that transneuronal transport had occurred. Electron microscopy confirmed transneuronal transport between olfactory axons and second order bulbar neurons. These results show that the capacity for olfactory neuron recovery and reconnection persists in the hamster well into old age.