Studies of royal jelly and associated cross-reactive allergens in atopic dermatitis patients.

Royal jelly (RJ), a creamy substance secreted by honeybees, is the exclusive diet for queen bee differentiation and life maintenance. RJ has been used in cosmetics, beverages, medicines, and supplements worldwide. However, allergy is a concerning issue for RJ, especially in atopic dermatitis (AD) and asthma patients. In some cases, allergic reactions are seen after the first intake of RJ, suggesting the existence of allergens cross-reactive with RJ. Information about the cross-reactive allergens is very important for the safe application of RJ; however, study of this cross-reactivity is quite limited. In this study, we attempted to identify allergens cross-reactive with RJ by using serum samples from 30 AD patients who had never been exposed to RJ. In an enzyme-linked immunosorbent assay (ELISA) experiment, RJ-binding IgE antibodies were detected in the serum of 10 out of 30 patients, and their antibody titers ranged from 4- to 2,048-fold dilution ratios. Additionally, 3 AD patients were determined to be positive in a skin-prick test (SPT) with an RJ solution. Significant correlations were observed between the anti-RJ antibody titer and nonspecific IgE and between the anti-RJ antibody titer and the Eczema Area and Severity Index score. We further examined the cross-reactivity between RJ and 14 typical allergens by using an ELISA-inhibition assay and demonstrated that the following 6 allergens showed cross-reactivity with RJ: the European house dust mite (HDM) (Dermatophagoides pteronyssinus), American HDM (Dermatophagoides farinae), snow crab (Chionocetes spp.), edible crab (Cancer pagurus), German cockroach (Blatella germanica), and honeybee venom (Apis mellifera). In conclusion, people with a history of allergic diseases, including AD, asthma, and allergic rhinitis, should be cautioned against consuming RJ products because of the potential for cross-reactive responses to ensure the safe and successful use of RJ supplements.

Chemical constituents of Brazilian Propolis from the state of Bahia and their growth inhibitory activities against cancer cells.

A novel 2-phenoxychromone (1) and five known flavones (2-6) were isolated from northeastern Brazilian propolis in the state of Bahia. The chemical structures of these six compounds were determined by spectroscopic investigations and single-crystal X-ray analysis. The isolated compounds showed growth-inhibitory activities, in varying degrees, against human tumor cell lines. This is the first report on the discovery of a novel 2-phenoxychromone from propolis.

Quantitative Analysis of Apisin, a Major Protein Unique to Royal Jelly.

Apisin, a protein that is unique to royal jelly (RJ), is known to compose the greater part of the RJ proteins and to exist as a heterooligomer containing major royal jelly protein 1 and apisimin. However, few reports on the methods for quantifying apisin have been published. Thus, we attempted to quantify apisin using HPLC, a widely used analytical technique, as described below. Isoelectric precipitation and size-exclusion chromatography were used to obtain the purified protein, which was identified as apisin by SDS-PAGE and LC-MS analyses. The purified apisin was lyophilized and then used to generate a calibration curve to quantify apisin in RJ. The apisin content was fairly constant (i.e., 3.93 to 4.67 w/w%) in natural RJ. This study is the first to describe a simple, standardized method for quantifying apisin using HPLC and suggests that apisin can be used as a benchmark for future evaluations of RJ quality.

Discovery of a Novel Diterpene in Brown Propolis from the State of Parana, Brazil.

Propolis is a resinous substance collected by honeybees from certain plant sources. The components of propolis depend on the vegetation of the area in which apiculture is practiced. In Brazil, there are several types of propolis including 'green,' 'red' and 'brown'. Brazilian brown propolis from the state of Parana characteristically includes diterpenes, and we discovered a novel clerodane diterpene, rel-(5S,6S,8R,9R,10S,18R,19S)-18,19-epoxy-2-oxocleroda-3,12(E),14- triene-6,18,19-triol 18,19-diacetate 6-benzoate (3) and five known diterpenes (1, 2, 4, 5 and 6). The chemical structure of the novel diterpene 3 was determined using 1D- and 2D-NMR spectroscopic analyses. Furthermore, the activities of the isolated diterpenes on growth inhibition of several human cancer cell lines (LNCaP, MCF-7, DLD-1 and A549) were evaluated in vitro; diterpene 3 exhibited a potent inhibition of cell growth, and its activity was approximately 15 times higher than that of the other diterpenes.

Dual-energy subtraction radiography improves laryngeal delineation in patients with moderate to severe cervical spondylosis.

PURPOSE: To investigate the feasibility of dual-energy subtraction (DES) in patients with moderate-severe cervical spondylosis for improving delineation of the larynx on flat panel detector (FPD) radiography. MATERIALS AND METHODS: For 118 patients, we graded conventional/DES anterior-posterior views for delineation of the vocal cords, subglottis, and pyriform sinus using a 5-point scale and lateral views from conventional laryngeal FPD radiography to determine cervical spondylosis severity on a scale from 0 (none) to 3 (severe). We compared the delineation of each anatomical structure in both groups of grades 0-1 and grades 2-3 of spondylosis severity between conventional and DES methods and the improved delineation rate for each anatomical structure by DES compared to the conventional method between both groups. RESULTS: With DES, the delineation of each anatomical structure was significantly better than with conventional radiography for both groups (P < 0.0001). The improved delineation rate of the vocal cord and subglottis using DES was significantly higher in grades 2-3 than in grades 0-1 (P < 0.05), although there was no significant difference in the delineation rate of the pyriform sinus between the groups (P = 0.847). CONCLUSION: DES provides better delineation of the laryngeal anatomy than conventional FPD radiography predominantly in patients with moderate-severe cervical spondylosis.

Planer orientation of the bilateral semicircular canals in dizzy patients.

OBJECTIVE: Recent development of 3-dimensional analysis of eye movement enabled to detect the eye rotation axis, which is used to determine the responsible semicircular canal(s) in dizzy patients. Therefore, the knowledge of anatomical orientation of bilateral semicircular canals is essential, as all 6 canals influence the eye movements. SUBJECTS AND METHODS: Employing the new head coordinate system suitable for MR imaging, we calculated the angles of semicircular canal planes of both ears in 11 dizzy patients who had normal caloric response in both ears. RESULTS: The angles between adjacent canal pairs were nearly perpendicular in both ears. The angle between the posterior canal planes and head sagittal plane was 51° and significantly larger the angle between the anterior canal planes and head sagittal plane, which was 35°. The angle between the horizontal canal plane and head sagittal plane was almost orthogonal. Pairs of contralateral synergistic canal planes were not parallel, forming 10° between right and left horizontal canal planes, 17° between right anterior and left posterior canal planes and 19° between the right posterior and left anterior canal planes. CONCLUSION: Our measurement of the angles of adjacent canal pairs and the angle between each semicircular canal and head sagittal plane coincided with those of previous reports obtained from CT images and skull specimens. However, the angles between contralateral synergistic canal planes were more parallel than those of previous reports.

Spatial orientation of the angular vestibulo-ocular reflex (aVOR) after semicircular canal plugging and canal nerve section.

We investigated spatial responses of the aVOR to small and large accelerations in six canal-plugged and lateral canal nerve-sectioned monkeys. The aim was to determine whether there was spatial adaptation after partial and complete loss of all inputs in a canal plane. Impulses of torques generated head thrusts of ≈ 3,000°/s². Smaller accelerations of ≈ 300°/s² initiated the steps of velocity (60°/s). Animals were rotated about a spatial vertical axis while upright (0°) or statically tilted fore-aft up to ± 90°. Temporal aVOR yaw and roll gains were computed at every head orientation and were fit with a sinusoid to obtain the spatial gains and phases. Spatial gains peaked at ≈ 0° for yaw and ≈ 90° for roll in normal animals. After bilateral lateral canal nerve section, the spatial yaw and roll gains peaked when animals were tilted back ≈ 50°, to bring the intact vertical canals in the plane of rotation. Yaw and roll gains were identical in the lateral canal nerve-sectioned monkeys tested with both low- and high-acceleration stimuli. The responses were close to normal for high-acceleration thrusts in canal-plugged animals, but were significantly reduced when these animals were given step stimuli. Thus, high accelerations adequately activated the plugged canals, whereas yaw and roll spatial aVOR gains were produced only by the intact vertical canals after total loss of lateral canal input. We conclude that there is no spatial adaptation of the aVOR even after complete loss of specific semicircular canal input.

Dual-energy subtraction imaging for diagnosing vocal cord paralysis with flat panel detector radiography.

OBJECTIVE: To investigate the clinical feasibility of dual energy subtraction (DES) imaging to improve the delineation of the vocal cord and diagnostic accuracy of vocal cord paralysis as compared with the anterior-posterior view of flat panel detector (FPD) neck radiography. MATERIALS AND METHODS: For 122 consecutive patients who underwent both a flexible laryngoscopy and conventional/DES FPD radiography, three blinded readers retrospectively graded the radiographs during phonation and inspiration on a scale of 1 (poor) to 5 (excellent) for the delineation of the vocal cord, and in consensus, reviewed the diagnostic accuracy of vocal cord paralysis employing the laryngoscopy as the reference. We compared vocal cord delineation scores and accuracy of vocal cord paralysis diagnosis by both conventional and DES techniques using kappa statistics and assessing the area under the receiver operating characteristic curve (AUC). RESULTS: Vocal cord delineation scores by DES (mean, 4.2 +/- 0.4) were significantly higher than those by conventional imaging (mean, 3.3 +/- 0.5) (p < 0.0001). Sensitivity for diagnosing vocal cord paralysis by the conventional technique was 25%, whereas the specificity was 94%. Sensitivity by DES was 75%, whereas the specificity was 96%. The diagnostic accuracy by DES was significantly superior (kappa = 0.60, AUC = 0.909) to that by conventional technique (kappa = 0.18, AUC = 0.852) (p = 0.038). CONCLUSION: Dual energy subtraction is a superior method compared to the conventional FPD radiography for delineating the vocal cord and accurately diagnosing vocal cord paralysis.

A new coordinates system for cranial organs using magnetic resonance imaging.

CONCLUSION: We developed a new coordinates system for magnetic resonance imaging (MRI) that utilizes the labyrinth and eyeballs as references to measure the spatial arrangement of cranial organs, and we verified its usefulness by observing small structures in the labyrinth in 39 ears from 33 patients. Our new coordinates system could be used for stereotactic analysis of cranial organs in MRI. OBJECTIVES: To research semicircular canal anatomy in healthy organisms, we propose a method that employs references visible on MRI for stereotactic measurement of cranial structures, and we evaluated the usefulness of our method. METHODS: Using the new coordinates system and vector analysis, we calculated angles among the semicircular canals and sagittal head plane from MRI volume data containing temporal bone and orbit. RESULTS: The angle between the anterior semicircular canal plane and sagittal plane was 35.3 +/- 4.1 degrees; posterior semicircular canal plane and sagittal plane, 50.9 +/- 4.7 degrees; and horizontal semicircular canal plane and sagittal plane, 90.4 +/- 7.0 degrees. The angle between the anterior and posterior semicircular canal planes was 95.1 +/- 4.2 degrees; anterior and horizontal semicircular canal planes, 92.3 +/- 7.5 degrees; and posterior and horizontal semicircular canal planes, 93.5 +/- 4.9 degrees.

A head-tilt caloric test for evaluating the vertical semicircular canal function.

CONCLUSIONS: The caloric test with head-tilt can be used as a tool for assessing vertical canal function as an office procedure. OBJECTIVE: Evaluation of vertical canal function. PATIENTS AND METHODS: We provoked caloric response by cold water in the vertiginous patients in supine position. During the culmination of the response we rotated the head 45 degrees from the sagittal plane to place the posterior canal to earth-vertical. Thereafter we rotated the head 45 degrees to the opposite direction to place the anterior canal to earth-vertical. The eye movements were recorded by two-dimensional electronystagmography. The data collected from the examination of 100 ears with normal caloric response in horizontal component were analyzed. RESULTS: The down-beating vertical component intensified when the posterior canal was placed to earth-vertical. The up-beating vertical component intensified when the anterior canal was placed to earth-vertical. These findings suggested that the vertical canals were functioning.

Horizontal saccadic palsy associated with gliosis of the brainstem midline.

We studied premotor cell groups involved in the generation of saccades in a patient with a disturbance of voluntary horizontal gaze. The only neurological symptom found was a slowing of horizontal saccades, reported since birth and unaltered over his lifetime. We attribute this disorder, for the first time, to a fibrous gliosis of the brainstem midline, which may disrupt neuronal elements of the horizontal saccade generator crossing the brainstem midline, but it caused no obvious loss of omnipause-, excitatory burst-, and inhibitory burst neurons. No neuronal loss or demyelination, was apparent elsewhere in the brainstem; but there was evidence of an ependymal infection throughout the entire ventricular system. A diagnosis of Gaucher disease was made from the bone marrow of this patient shortly before his death, but for several reasons we considered this complication unlikely to be the cause of the saccadic disorder.

An effective maneuver of positional test by turning of the head and body together.

OBJECTIVE: To find the effective maneuver of a positional test. METHODS: We performed three different maneuvers of a positional test in 86 vertiginous patients in supine-lying position in the following order: (1) "body-only maneuver": a patient turned only his body to one side while keeping his head still; (2) "head-only maneuver": a patient turned only his head to one side while keeping the body still; (3) "head-and-body maneuver": a patient turned his head and body together to one side. Eye movement during the whole procedure was recorded by two-dimensional electronystagmography. RESULTS: Nystagmus provocation rates were 9% in "body-only maneuver", 16% in "head-only maneuver", and 33% in "head-and-body maneuver". CONCLUSION: The positional test in which patient turned his head and body together proved most effective.

Clinical significance of vertical component of caloric response including its second phase in vertiginous patients.

CONCLUSIONS: Up-beating vertical component recorded in the caloric first phase was attributed mainly to the inhibitory endolymph flow in the anterior canal. Down-beating vertical component recorded in the caloric second phase provoked by a positional change could be explained by a reversed endolymph flow in vertical canal(s). OBJECTIVE: To investigate the origin of a vertical component in caloric response. MATERIALS AND METHODS: We analyzed electronystagmography (ENG) of caloric responses, which had measurable horizontal component in the caloric first phase in both ears in 200 ears of 100 vertiginous patients. A caloric first phase was provoked by cold water in the supine position with the lateral semicircular canal earth-vertical. A caloric second phase was provoked by re-orienting the lateral canal from the earth-vertical to earth-horizontal after the cessation of the first phase (provoked second phase). The nystagmus of the whole procedure was recorded by two-dimensional ENG. RESULTS: We recorded the vertical component in 103/200 ears in the caloric first phase, which was directed mostly upward (92/103 ears). We also recorded the vertical component in 91/200 ears in the provoked second phase, which was directed almost exclusively downward (90/91 ears).

Effective maneuver of the positional test: turning head and body together.

We performed three maneuvers of the positional test in supine patients: (1) body maneuver, in which patients turned only the body to one side while keeping the head as still as possible; (2) head-only maneuver, in which patients turned only the head to one side while keeping the body still; and (3) head and body maneuver, in which patients turned the head and body together to one side. The nystagmus provocation rates of those three maneuvers among 86 vertiginous patients were 9% in the body maneuver, 16% in the head-only maneuver, and 33% in the head and body maneuver, respectively. We proved that performing the positional test by turning the head and body together is very effective in provoking positional nystagmus.

Monocular electronystagmographic analysis of caloric nystagmus.

We compared the horizontal component of nystagmus of the right and left eyes using monocular recording of electronystagmography. We examined the eye movements of 135 patients during bithermal caloric testing and those of 50 patients during the rotation test. We measured the number of nystagmic beats, the slow-phase velocities, and the amplitudes during 10 seconds of the culmination phase of caloric response. We also measured the number of nystagmic beats during the first 30 seconds in postrotatory nystagmus. The eye on the cold-irrigated side moved significantly more strongly than did the eye on the nonirrigated side, whereas a warm irrigation did not induce a significant difference between the irrigated and nonirrigated eyes. The summated activities of each eye during the four different stimulations under bithermal caloric testing did not show any significant differences. The activities of postrotatory nystagmus were almost equal in both eyes in 50 patients. We concluded that the inhibitory effect of cold caloric stimulation is probably transmitted more intensively to the eye on the irrigated side.

Vertical component of the caloric response, including a caloric second phase provoked by positional change: a preliminary report.

We measured the horizontal and vertical components of caloric nystagmus in 120 ears of 60 vertiginous patients who had moderate to vigorous caloric first-phase response in both ears, no spontaneous nystagmus, and no severe disorders in the central nervous system. We provoked a caloric first phase in the supine position with 5 ml of water at 20 degrees C for 15 seconds. We provoked a caloric second phase by changing the position of a patient from supine to sitting after the end of the first phase. The horizontal component of the caloric second phase was recorded in 108 of 120 ears (90%). The vertical component was recorded in 57 of 120 ears (48%) during the caloric first phase and in 51 of 120 ears (43%) during the caloric second phase. We suspected that the vertical component of the caloric first phase was mainly due to the inhibition of the anterior semicircular canal.

The clinical significance of the caloric second phase provoked by positional change in vertiginous patients.

The reversal phase of caloric nystagmus is provoked when the lateral semicircular canal in a patient is reoriented from a vertical to a horizontal plane at the cessation of the caloric first phase, which we called the provoked caloric second phase. In investigating the clinical significance of the provoked caloric second phase, we recruited 102 vertiginous patients who had measurable caloric responses in both ears but no disorders of the central nervous system. We recorded the provoked caloric second phase in 188 (92%) of 204 ears in 102 patients. The average maximum slow-phase velocity of the caloric first phase was 26.9 degrees per second, and that of the provoked caloric second phase was 5.0 degrees per second. The maximum slow-phase velocity of the provoked caloric second phase correlated with that of the foregoing caloric first phase (r = -.84). Thus, we consider that the provoked caloric second phase is influenced largely by the foregoing caloric first phase. Furthermore, in the patients who responded normally to caloric stimulation, the directional preponderance of the provoked caloric second phase correlated with the directional preponderance of optokinetic after-nystagmus (r = .64). Hence, we conclude that the provoked caloric second phase reflects central vestibular asymmetry in patients with normal peripheral vestibular function.

Spatial orientation of caloric nystagmus in semicircular canal-plugged monkeys.

We studied caloric nystagmus before and after plugging all six semicircular canals to determine whether velocity storage contributed to the spatial orientation of caloric nystagmus. Monkeys were stimulated unilaterally with cold ( approximately 20 degrees C) water while upright, supine, prone, right-side down, and left-side down. The decline in the slow phase velocity vector was determined over the last 37% of the nystagmus, at a time when the response was largely due to activation of velocity storage. Before plugging, yaw components varied with the convective flow of endolymph in the lateral canals in all head orientations. Plugging blocked endolymph flow, eliminating convection currents. Despite this, caloric nystagmus was readily elicited, but the horizontal component was always toward the stimulated (ipsilateral) side, regardless of head position relative to gravity. When upright, the slow phase velocity vector was close to the yaw and spatial vertical axes. Roll components became stronger in supine and prone positions, and vertical components were enhanced in side down positions. In each case, this brought the velocity vectors toward alignment with the spatial vertical. Consistent with principles governing the orientation of velocity storage, when the yaw component of the velocity vector was positive, the cross-coupled pitch or roll components brought the vector upward in space. Conversely, when yaw eye velocity vector was downward in the head coordinate frame, i.e., negative, pitch and roll were downward in space. The data could not be modeled simply by a reduction in activity in the ipsilateral vestibular nerve, which would direct the velocity vector along the roll direction. Since there is no cross coupling from roll to yaw, velocity storage alone could not rotate the vector to fit the data. We postulated, therefore, that cooling had caused contraction of the endolymph in the plugged canals. This contraction would deflect the cupula toward the plug, simulating ampullofugal flow of endolymph. Inhibition and excitation induced by such cupula deflection fit the data well in the upright position but not in lateral or prone/supine conditions. Data fits in these positions required the addition of a spatially orientated, velocity storage component. We conclude, therefore, that three factors produce cold caloric nystagmus after canal plugging: inhibition of activity in ampullary nerves, contraction of endolymph in the stimulated canals, and orientation of eye velocity to gravity through velocity storage. Although the response to convection currents dominates the normal response to caloric stimulation, velocity storage probably also contributes to the orientation of eye velocity.

Langerhans cells within the follicular epithelium and the intradermal sweat duct in equine insect hypersensitivity "Kasen".

Histopathologic and electron microscopic observations were given on Langerhans cells (LCs) within the follicular epithelium (FE) and intradermal sweat duct (ISD) of equine "Kasen". By light microscopy, LCs were present in the greatest numbers within the FE and ISD than within the epidermal layer and the normal skin, with an occasional formation of several aggregated foci. By electron microscopy, LCs within the FE and ISD widely extended their dendritic processes between the keratinocytes and contained Birbeck granules (Bgs), mitochondria, rough endoplasmic reticula and ribosomes in the cytoplasm. Numerous Type 2 LCs, with a number of Bgs and endocytosis, and Type 3 LCs, with multivesicular bodies and endosomes of various sizes, were recognized within the FE and ISD, although inactive Type 1 LCs, with a narrow and lucid cytoplasm, were rarely seen. LCs observed within the FE and ISD in the "Kasen" skin lesions might express the particular stage corresponded to recognize, intake and process the antigens which permeate them.

[Evaluation of safe surgical treatment of peritonsillar abscess using computed tomography].

With the development of new antimicrobial agents, the incidence of peritonsillar abscess (PTA) is on the decline. PTA is still often encountered in general practice, however, where it requires immediate diagnosis and treatment. Because the internal carotid artery runs medially to the medial parapharyngeal space, damage to nearby vascular or other structures is a surgical risk of PTA. We used contrast computed tomography (CT) from PTA patients to investigate the anatomical relationship between the abscess and parapharyngeal space, and to determine safe surgical sites. We observed 31 patients with PTA--19 men and 12 women--between February 1997 and April 1999, all examined by contrast CT and undergoing drainage or incision. The average age was 30.7 years (range: 12-54 years). The abscess was on the right side in 20 cases and on the left side in 11. We determined the sites of the abscess and carotid artery, internal jugular vein, and surrounding soft tissue density area including nerves in the parapharyngeal space based on the angle and distance from recognizable anatomical structures in CT scans. The anterior margin of the parapharyngeal space was 29 +/- 5 mm posterior from the upper posterior alveolar margin. The medial margin of that space was at 15 +/- 2 degrees laterally from the midline of the incisors, and 24 +/- 4 mm laterally from the midline sagittal plane. The internal carotid artery was located medially to the parapharyngeal space, running on the sagittal plane containing the upper posterior alveolar margin. The distance from the anterior margin of the parapharyngeal space to the posterior wall of the PTA was 9 +/- 4 mm, and the distance to the anterior wall of the abscess (including the pharyngeal mucosa) was 31 +/- 5 cm. The relationship between the upper posterior alveolar margin and midline sagittal plane was useful for determining the site of the parapharyngeal space. Because the internal carotid artery is located on the same sagittal plane as the upper posterior alveolar margin, when conducting drainage or incision of PTA, we should advance sagittaLly from the point of incision to a depth of no more than 20 mm. If the tip of the instrument is kept medial to the sagittal plane of the upper posterior alveolar margin, effective treatment should be achievable without the risk of vascular damage.