Innervation of the Human Incisive Papilla: Comparison with Other Oral Regions.

Immunohistochemistry for several neurochemical substances was performed on the human incisive papilla and other oral structures. Sodium channel alpha subunit 7 (SCN7A) protein-immunoreactive (IR) Schwann cells and protein gene product 9.5 (PGP 9.5)-IR nerve fibers made nerve plexuses beneath the epithelium of the palate, including the incisive papilla, tongue, and lip. SCN7A immunoreactivity could also be detected in lamellated and nonlamellated capsules of corpuscle endings. Lamellated SCN7A-IR corpuscle endings were mostly restricted to the mucous and cutaneous lips. These endings had thick and spiral-shaped PGP 9.5-IR axons without ramification. Nonlamellated SCN7A-IR corpuscle endings were most numerous in the incisive papilla among the oral regions. On the basis of axonal morphology, the nonlamellated endings were divided into simple and complex types. PGP 9.5-IR terminal axons in the simple type ran straight or meandered with slight ramification, whereas those in the complex type were densely entangled with abundant ramification. Substance P (SP)-, calcitonin gene-related peptide (CGRP)-, and transient receptor potential cation channel subfamily V member 2 (TRPV2)-IR varicose fibers were rarely seen beneath the epithelium of oral structures. The present study indicates that the human incisive papilla has many low-threshold mechanoreceptors with nonlamellated capsules. SP-, CGRP-, and TRPV2-containing nociceptors may be infrequent in the incisive papilla and other oral regions.

TRPC1, TRPC3, and TRPC4 in Rat Orofacial Structures.

TRPC (transient receptor potential cation channel subfamily C) members are nonselective monovalent cation channels and control Ca2+ inflow. In this study, immunohistochemistry for TRPC1, TRPC3, and TRPC4 was performed on rat oral and craniofacial structures to elucidate their distribution and function in the peripheries. In the trigeminal ganglion (TG), 56.1, 84.1, and 68.3% of sensory neurons were immunoreactive (IR) for TRPC1, TRPC3, and TRPC4, respectively. A double immunofluorescence method revealed that small to medium-sized TG neurons co-expressed TRPCs and calcitonin gene-related peptide. In the superior cervical ganglion, all sympathetic neurons showed TRPC1 and TRPC3 immunoreactivity. Parasympathetic neurons in the submandibular ganglion, tongue, and parotid gland were TRPC1, TRPC3, and TRPC4 IR. Gustatory and olfactory cells were also IR for TRPC1, TRPC3, and/or TRPC4. In the musculature, motor endplates expressed TRPC1 and TRPC4 immunoreactivity. It is likely that TRPCs are associated with sensory, autonomic, and motor functions in oral and craniofacial structures.

Sensory Neurons in the Human Geniculate Ganglion.

The geniculate ganglion (GG) contains visceral and somatic sensory neurons of the facial nerve. In this study, the number and cell size of sensory neurons in the human GG were investigated. The estimated number of GG neurons ranged from 1,580 to 2,561 (mean ± SD = 1,960 ± 364.6). The cell size of GG neurons ranged from 393.0 to 2,485.4 µm2 (mean ± SD = 1,067.4 ± 99.5 µm2). Sensory neurons in the GG were significantly smaller in size than those in the dorsal root (range = 326.6-5343.4 µm2, mean ± SD = 1,683.2 ± 203.8 µm2) or trigeminal ganglia (range = 349.6-4,889.28 µm2, mean ± SD = 1,529.0 ± 198.48 µm2). Sensory neurons had similar cell body sizes in the GG and nodose ganglion (range = 357.2-3,488.33 µm2, mean ± SD = 1,160.4 ± 156.61 µm2). These findings suggest that viscerosensory neurons have smaller cell bodies than somatosensory neurons. In addition, immunohistochemistry for several neurochemical substances was performed on the human GG. In the ganglion, sensory neurons were mostly immunoreactive for secreted protein, acidic and rich in cysteine-like 1 (94.3%). One third of GG neurons showed vesicular glutamate transporter 2 immunoreactivity (31.3%). Only 7.3% of GG neurons were immunoreactive for transient receptor potential cation channel subfamily V member 1. Sensory neurons in the human GG may respond to gustatory, nociceptive, and/or mechanoreceptive stimuli from tongues, soft palates, and external auditory canals.

Distribution of CGRP and TRPV2 in Human Paranasal Sinuses.

Immunohistochemistry for protein gene product 9.5 (PGP 9.5), calcitonin gene-related peptide (CGRP) and the transient receptor potential cation channel subfamily V member 2 (TRPV2) was performed on human paranasal sinuses. It was found that in the paranasal sinuses, mucous membranes contain PGP 9.5-immunoreactive (PGP 9.5-IR) nerve fibers. Such nerve fibers terminated around large blood vessels as fine varicosities. Isolated PGP 9.5-IR nerve fibers were scattered beneath the epithelium. Glandular tissues were also innervated by PGP 9.5-IR nerve fibers. These fibers were numerous in the maxillary and ethmoid sinuses, and relatively rare in the frontal and sphenoid sinuses. CGRP-IR nerve fibers were common in the maxillary sinus whereas TRPV2-IR nerve fibers were abundant in the ethmoid sinus. They were located around large blood vessels in the lamina propria. Many subepithelial nerve fibers contained TRPV2 immunoreactivity in the ethmoid sinus. CGRP- and TRPV2-IR nerve fibers were very infrequent in the frontal and sphenoid sinuses. In the human trigeminal ganglion (TG), sensory neurons contained CGRP or TRPV2 immunoreactivity. CGRP-IR TG neurons were more common than TRPV2-IR TG neurons. CGRP-IR TG neurons were of various cell body sizes, whereas TRPV2-IR TG neurons were mostly medium-to-large. In addition, human spinal and principal trigeminal sensory nuclei contained abundant CGRP- and TRPV2-IR varicosities. This study indicates that CGRP- and TRPV2-containing TG neurons probably innervate the paranasal sinus mucosae, and project into spinal and principal trigeminal sensory nuclei.

SCN2B in the Rat Trigeminal Ganglion and Trigeminal Sensory Nuclei.

The beta-2 subunit of the mammalian brain voltage-gated sodium channel (SCN2B) was examined in the rat trigeminal ganglion (TG) and trigeminal sensory nuclei. In the TG, 42.6 % of sensory neurons were immunoreactive (IR) for SCN2B. These neurons had various cell body sizes. In facial skins and oral mucosae, corpuscular nerve endings contained SCN2B-immunoreactivity. SCN2B-IR nerve fibers formed nerve plexuses beneath taste buds in the tongue and incisive papilla. However, SCN2B-IR free nerve endings were rare in cutaneous and mucosal epithelia. Tooth pulps, muscle spindles and major salivary glands were also innervated by SCN2B-IR nerve fibers. A double immunofluorescence method revealed that about 40 % of SCN2B-IR neurons exhibited calcitonin gene-related peptide (CGRP)-immunoreactivity. However, distributions of SCN2B- and CGRP-IR nerve fibers were mostly different in facial, oral and cranial structures. By retrograde tracing method, 60.4 and 85.3 % of TG neurons innervating the facial skin and tooth pulp, respectively, showed SCN2B-immunoreactivity. CGRP-immunoreactivity was co-localized by about 40 % of SCN2B-IR cutaneous and tooth pulp TG neurons. In trigeminal sensory nuclei of the brainstem, SCN2B-IR neuronal cell bodies were common in deep laminae of the subnucleus caudalis, and the subnuclei interpolaris and oralis. In the mesencephalic trigeminal tract nucleus, primary sensory neurons also exhibited SCN2B-immunoreactivity. In other regions of trigeminal sensory nuclei, SCN2B-IR cells were very infrequent. SCN2B-IR neuropil was detected in deep laminae of the subnucleus caudalis as well as in the subnuclei interpolaris, oralis and principalis. These findings suggest that SCN2B is expressed by various types of sensory neurons in the TG. There appears to be SCN2B-containing pathway in the TG and trigeminal sensory nuclei.

SPARCL1-containing neurons in the human brainstem and sensory ganglion.

Secreted protein, acidic and rich in cysteine-like 1 (SPARCL1) is a member of the osteonectin family of proteins. In this study, immunohistochemistry for SPARCL1 was performed to obtain its distribution in the human brainstem, cervical spinal cord, and sensory ganglion. SPARCL1-immunoreactivity was detected in neuronal cell bodies including perikarya and proximal dendrites, and the neuropil. The motor nuclei of the IIIrd, Vth, VIth, VIIth, IXth, Xth, XIth, and XIIth cranial nerves and spinal nerves contained many SPARCL1-immunoreactive (-IR) neurons with medium-sized to large cell bodies. Small and medium-sized SPARCL1-IR neurons were distributed in sensory nuclei of the Vth, VIIth, VIIIth, IXth, and Xth cranial nerves. In the medulla oblongata, the dorsal column nuclei also had small to medium-sized SPARCL1-IR neurons. In addition, SPARCL1-IR neurons were detected in the nucleus of the trapezoid body and pontine nucleus within the pons and the arcuate nucleus in the medulla oblongata. In the cervical spinal cord, the ventral horn contained some SPARCL1-IR neurons with large cell bodies. These findings suggest that SPARCL1-containing neurons function to relay and regulate motor and sensory signals in the human brainstem. In the dorsal root (DRG) and trigeminal ganglia (TG), primary sensory neurons contained SPARCL1-immunoreactivity. The proportion of SPARCL1-IR neurons in the TG (mean ± SD, 39.9 ± 2.4%) was higher than in the DRG (30.6 ± 2.1%). SPARCL1-IR neurons were mostly medium-sized to large (mean ± SD, 1494.5 ± 708.3 µm(2); range, 320.4-4353.4 µm(2)) in the DRG, whereas such neurons were of various cell body sizes in the TG (mean ± SD, 1291.2 ± 532.8 µm(2); range, 209.3-4326.4 µm(2)). There appears to be a SPARCL1-containing sensory pathway in the ganglion and brainstem of the spinal and trigeminal nervous systems.

[Clinical Characteristics of Adult Onset RSV Infection During Two Consecutive Winter Seasons].

We performed a retrospective study examining adult patients with RSV infection who were diagnosed at our hospital during two consecutive winter seasons, 2011-2012 and 2012-2013; these patients were compared with, adult patients who had been diagnosed as having influenza during the same periods. RSV infection was confirmed by a 4-fold increase in the CF antibody titer, while influenza was diagnosed based on a rapid antigen test. Forty-three patients with RSV infection and pneumonia patients (39 inpatients and 4 outpatients) and 25 patients with influenza and pneumonia (23 inpatients and 2 outpatients) were detected. Overall, 54 patients with RSV infection and 42 patients with influenza, were hospitalized during the two seasons. A history of the influenza vaccination was verified for 48% of the influenza patients with pneumonia and 35% of the non-pneumonia influenza patients who were hospitalized, and neuraminidase inhibitors were used for the treatment of all the influenza patients and 88% of the non-pneumonia influenza patients who were hospitalized. Overall, 5.3% of the adult cases with pneumonia (43/817) during the two seasons were diagnosed as having RSV related illness, and within the peak period, in particular, 14.6% were judged as having RSV pneumonia. Furthermore, 63% of the patients with RSV infection and pneumonia had mixed infections with other common respiratory pathogen, such as Streptococcus pneumoniae, and within the peak period, almost 15% of the patients with pneumonococcal pneumonia were confirmed to have mixed infections that included RSV. In both groups, one-fourth of the patients had been living in nursing homes or had been receibing home medical care. Up to 20% of the RSV pneumonia patients were initially diagnosed as having aspiration pneumonia. We suspect that some of these elderly patients might have developed pneumonia as a result of preceding viral infection or following vomiting or aspiration. The overall clinical picture, such as the mean age, maximum body temperature, hypoxemia, CRP, and WBC, did not differ significantly between the two groups. The 30-day mortality and overall hospital mortality rates were similar in both groups, but the lengths of the hospital stay were significantly longer, and several patients survived but continued to have a reduced activities of daily living score at the time of their discharge in the RSV pneumonia group.

[[TUBERCULOSIS DIAGNOSIS FOLLOWING A VISIT TO THE EMERGENCY ROOM].]

We aimed to evaluate the clinical char- acteristics of patients admitted to the emergency room (ER) and diagnosed with tuberculosis. [Method] We conducted a retrospective study of patients aged ; 16 years admitted to the hospital between April 1980 and March 2015 and diagnosed with tuberculosis. We com- pared patient clinical characteristics and type of tuberculosis between ER and non-ER patients. We also compared the incidence of delayed diagnosis of tuberculosis between ER patients with and without respiratory symptoms. We compared the tuberculosis encounter rate and the time to diagnosis of tuberculosis in ER and non-ER patients. [Results] A total of 255 patients, including 54 ER and 201 non-ER patients were enrolled in this study. The average age J was higher in ER patients than in non-ER patients (71.7? 16.3 vs. 63.3 ?20.3 years, p=0.006). The reasons for visiting the ER included acute conditions such as fracture of the lumbar spine, acute myocardial infarction, hemorrhagic gastric ulcer, brain infarction, and carbon monoxide intoxication, requiring immediate treatment. The time to diagnosis of tuberculosis in ER patients without respiratory symptoms (n=21) was approximately three times longer than that in patients with respiratory symptoms (n=33) as urgent treatment is priori- tized. The tuberculosis encounter rate was 1/1,800 for pa- tients transported by ambulance and 1/22,000 for emergency outpatients. The time to diagnosis of tuberculosis for patients transported by ambulance was approximately 4-6 days lon- ger than that for emergency outpatients or non-ER patients. [Conclusion] Physicians should seek to rule out the possi- bility of tuberculosis in all patients admitted to the ER, even where more urgent clinical conditions are prioritized.

[Occurrence and clinical characteristics of tuberculosis among home medical care patients].

OBJECTIVE: To clarify the occurrence and clinical characteristics of tuberculosis among home medical care patients, we conducted a retrospective study of patients who received home medical care from our hospital. SUBJECTS AND METHODS: We investigated 502 patients (mean age, 79.5 years) who received home medical care from our hospital between January 2003 and December 2012. The newly notified tuberculosis cases aged > or = 70 years in the general population in Miyagi were defined as the control group. Among the patients receiving home medical care, we evaluated the clinical characteristics of the patients with tuberculosis. RESULTS: Four of the 502 patients (0.8%) developed tuberculosis. Using the person-years method, the case rate of tuberculosis was calculated as 298.3 per 100,000 among home medical care patients. Compared with the control group, home medical care patients had a greater incidence of tuberculosis (298.3 vs. 36.06; rate ratio, 8.27; 95% confidence interval, 3.06-22.3; p < 0.001). When home medical care patients visited the hospital or were transported there by ambulance, they were initially often diagnosed with aspiration pneumonia. Moreover, the time interval to the onset of disease from the introduction of home medical care varied among cases (3-192 months). CONCLUSION: Patients receiving home medical care are at high risk of contracting tuberculosis. Therefore, for the medical staff involved in treating home medical care patients, the onset of tuberculosis should be carefully considered in daily medical practice.

Distribution of alpha-synuclein in rat salivary glands.

Expression of alpha-synuclein (Syn), a presynaptic neuronal protein, was immunohistochemically examined in intact rat submandibular, sublingual, and lingual glands. The submandibular gland contained abundant periductal Syn-immunoreactive (-ir) nerve fibers. Abundant Syn-ir varicosities were present in acini of the sublingual and serous lingual glands. By confocal laser scanning microscopy, Syn-ir nerve fibers around smooth muscle actin (SMA)-ir cells alone were infrequent; however, those around aquaporin-5 (AQP5)-ir cells alone and both SMA- and AQP5-ir cells were abundant in the sublingual and serous lingual glands. SMA-ir cells were occasionally immunoreactive for toll-like receptor 4, a Syn receptor. Syn-ir nerve fibers contained tyrosine hydroxylase (TH) in the submandibular gland and choline acetyltransferase (ChAT) in all examined salivary glands. In the superior cervical (SCG), submandibular, and intralingual ganglia, sympathetic and parasympathetic neurons co-expressed Syn with TH and ChAT, respectively. SCG neurons innervating the submandibular gland contained mostly Syn. In the thoracic spinal cord, 14.7% of ChAT-ir preganglionic sympathetic neurons co-expressed Syn. In the superior salivatory nucleus, preganglionic parasympathetic neurons projecting to the lingual nerve co-expressed Syn and ChAT. The present findings indicate that released Syn acts on myoepithelial cells. Syn in pre- and post-ganglionic neurons may regulate neurotransmitter release and salivary volume and composition.

Atezolizumab-related sclerosing cholangitis with multiple liver abscesses in a patient with lung squamous cell carcinoma: A case report.

A 76-year-old man underwent an operation for lung squamous cell carcinoma in the right lower lobe, followed by initial adjuvant therapy with atezolizumab, an antibody against anti-programmed death-ligand 1 (PD-L1). On day 4 after atezolizumab treatment, the patient developed general malaise and fatigue. He was diagnosed with atezolizumab-induced sclerosing cholangitis. Steroid treatment was started, and patient's condition, including symptoms, laboratory data and imaging findings, improved. Antibiotic treatments were ended on day 40, and the steroid dose was gradually reduced. Multiple liver abscesses were observed on day 106, and another treatment with antibiotics became necessary. The patient eventually recovered from liver abscesses. Sclerosing cholangitis induced by immune checkpoint inhibitor is rare, and the long-term clinical data about this adverse effect is limited. Hence, we think it is important to raise an alarm over sclerosing cholangitis coupled with liver abscesses after immunosuppressive therapy.

Distribution and anti-nociceptive function of endomorphin-1 in the rat cranial sensory ganglia.

Distribution of endomorphin-1 (EM-1) was immunohistochemically investigated in the rat cranial sensory ganglia. Small to medium-sized neurons in the trigeminal (TG), petrosal (PG), and jugular ganglia (JG) expressed EM-1-immunoreactivity. However, EM-1-immunoreactive (-ir) neurons were infrequent in the nodose ganglion. In the brainstem, EM-1-ir varicose fibers were detected in the superficial layer of the medullary dorsal horn and the caudal part of the nucleus tractus solitarius. By trichrome immunofluorescence analysis, approximately 70% of EM-1-ir neurons were also immunoreactive for transient receptor potential vanilloid 1 (TRPV1) in all the examined ganglia. Additionally, 56.8% of EM1-ir TG neurons and approximately 30% of EM-1-ir PG and JG neurons showed calcitonin gene-related peptide (CGRP)-immunoreactivity. By a retrograde tracing method, several TG, PG, and JG neurons innervating the facial and external ear canal skin expressed EM-1-immunoreactivity. However, EM-1-ir neurons innervating the tooth pulp, circumvallate papilla, and pharynx were relatively rare. Thus, EM-1 expression and its coexistence with TRPV1 and CGRP in the cranial sensory neurons may depend on their various peripheral targets. EM1-ir neurons probably project to the superficial layer of the medullary dorsal horn and caudal part of the nucleus tractus solitarius. EM-1 may be involved in nociceptive transmission from the skin.

The distribution of TRPV1 and TRPV2 in the rat pharynx.

Immunohistochemistry for two nociceptive transducers, the transient receptor potential cation channel subfamily V members 1 (TRPV1) and 2 (TRPV2), was performed on the pharynx and its adjacent regions. TRPV1-immunoreactivity (IR) was detected in nerve fibers beneath and within the epithelium and/or taste bud-like structure. In the pharynx, these nerve fibers were abundant in the naso-oral part and at the border region of naso-oral and laryngeal parts. They were also numerous on the laryngeal side of the epiglottis and in the soft palate. TRPV2-IR was expressed by dendritic cells in the pharynx and epiglottis, as well as in the root of the tongue and soft palate. These cells were located in the epithelium and lamina propria. TRPV2-immunoreactive (IR) dendritic cells were numerous in the naso-oral part of the pharynx, epiglottis, and tongue. Abundance of TRPV2-IR dendritic processes usually obscured the presence of TRPV2-IR nerve fibers in these portions. However, some TRPV2-IR nerve fibers could be observed in the epithelium of the soft palate. Retrograde tracing method also revealed that sensory neurons which innervate the pharynx or soft palate were abundant in the jugular-petrosal ganglion complex and relatively rare in the nodose ganglion. In the jugular-petrosal ganglion complex, TRPV1- and TRPV2-IR were expressed by one-third of pharyngeal and soft palate neurons. TRPV2-IR was also detected in 11.5 % pharyngeal and 30.9 % soft palate neurons in the complex. Coexpression of TRPV1 and CGRP was frequent among pharyngeal and soft palate neurons. The present study suggests that TRPV1- and TRPV2-IR jugular-petrosal neurons may be associated with the regulation of the swallowing reflex.

[A case of septic pulmonary embolism due to Peptostreptococcus micros with multiple infection of the head and neck].

A 46-year-old man with no underlying diseases visited our hospital with otorrhea, ocular motility disorder of the left eye, dizziness and loss of appetite which had lasted for two months. Chest radiography and computed tomography (CT) showed bilateral multiple pulmonary nodules and cavities. Furthermore, CT of the head and neck revealed bilateral mastoiditis, a left orbital abscess and a deep neck abscess. Peptostreptococcus micros was cultured from blood and otorrhea specimens. In addition, P. micros DNA was detected with the polymerase chain reaction (PCR) method in the specimens from the site of culture-negative lesions (i.e. sputum, bronchoalveolar lavage, neck abscess). Thus, we diagnosed the lung lesions as septic pulmonary embolisms (SPEs). The clinical findings of the head and neck had improved following antibiotics treatment for five weeks, and follow-up chest radiography and CT showed that all lesions almost disappeared. Since some SPE patients demonstrate a slow progression, SPE should be included in the differential diagnosis of multiple pulmonary nodules such as Wegener's glanulomatosis or neoplasm.

The role of TRPV2 as a regulator on the osteoclast differentiation during orthodontic tooth movement in rats.

When orthodontic forces are applied to teeth, bone remodeling, which consists of bone resorption and bone formation, occurs around the teeth. Transient receptor potential vanilloid 2 (TRPV2) is a cation channel expressed in various cell types that responds to various stimuli, including mechanical stress, and involved in calcium oscillations during the early stages of osteoclast differentiation. However, in vivo expression of TRPV2 in osteoclasts has not yet been reported, and temporo-spatial expression of TRPV2 during osteoclast differentiation is unclear. In this study, we examined the TRPV2 expression during experimental tooth movement and assessed the effect of TRPV2 on osteoclast differentiation. TRPV2 was detected on day 1 after experimental tooth movement on the compression side, and the number of TRPV2-expressing cells significantly increased on day 7. These TRPV2-expressing cells had a single, or multiple nuclei and were positive for TRAP activity. Consistent with these in vivo findings, in vitro experiments using RAW264.7 osteoclast progenitor cells showed that TRPV2 mRNA was increased at the early stage of osteoclast differentiation and maintained until the late stage. Furthermore, a TRPV2 channel selective antagonist significantly inhibited osteoclast differentiation. These findings suggest that TRPV2 may have a regulatory role in osteoclast differentiation during orthodontic tooth movement.

Distribution and possible function of galanin about headache and immune system in the rat dura mater.

Galanin (GAL) is a nociceptive transmitter or modulator in the trigeminal sensory system. In this study, GAL expression was investigated in the rat dura mater to demonstrate its possible function in headache using immunohistochemical techniques. The cerebral falx and cerebellar dura mater received abundant blood and nerve supply, and were significantly thicker compared to other portions in the cerebral dura mater. GAL-immunoreactivity was expressed by cell and nerve fiber profiles. Presumed macrophages and dendritic cells contained GAL-immunoreactivity, and co-expressed with CD11b-immunoreactivity. Many isolated and perivascular nerve fibers also showed GAL-immunoreactivity. In addition, GAL-immunoreactive nerve fibers were present in the vicinity of macrophages and dendritic cells with either GAL- or ED1-immunoreactivity. GAL-immunoreactive cells and nerve fibers were common in the cerebral falx and cerebellar dura mater and infrequent in other portions. And, GAL-immunoreactive nerve fibers usually co-expressed calcitonin gene-related peptide (CGRP)-immunoreactivity. In the trigeminal ganglion, a substantial proportion of sensory neurons innervating the dura mater contained GAL-immunoreactivity (mean ± SD, 3.4 ± 2.2%), and co-expressed CGRP-immunoreactivity (2.7 ± 2.1%). The present study may suggest that GAL is associated with nociceptive transduction or modulation in the dura mater. GAL also possibly plays a role in the immune mechanism of the dura mater.

Transient receptor potential melastatin-7 in the rat dorsal root ganglion.

AIMS: Transient receptor potential melastatin-7 (TRPM7) is a selective cation permeable channel which plays important roles in cellular and developmental biology such as cell proliferation, survival, differentiation and migration. This channel is also known to be necessary for transmitter release in the peripheral nervous system. In this study, immunohistochemistry for TRPM7 was conducted in the rat lumbar dorsal root ganglion (DRG). METHODS: Triple immunofluorescence methods were used to demonstrate distribution of TRPM7 and its relationship to other TRP channels in the DRG. Retrograde tracing and double immunofluorescence methods were also performed to know peripheral targets of DRG neurons containing TRPM7 and TRP vanilloid 1 (TRPV1). In addition, transection of the sciatic nerve was conducted to demonstrate an effect of the nerve injury on TRPM7expression in the DRG. RESULTS: TRPM7-immunoreactivity was expressed by 53.9% of sensory neurons in the 4th lumbar DRG. TRPM7-immunoreactive (-IR) DRG neurons mostly had small (<600 µm²) and medium-sized (600-1200 µm²) cell bodies. By triple and double immunofluorescence methods, approximately 70% of TRPM7-IR DRG neurons contained TRPV1-immunoreactivity. Although the number of DRG neurons co-expressing TRPM7 and TRPM8 was small in the DRG, almost all of TRPM8-IR DRG neurons co-expressed TRPM7-immunoreactivity. By combination of retrograde tracing method and immunohistochemistry, TRPM7 was expressed by half of DRG neurons innervating the plantar skin (61.9%) and gastrocnemius muscle (51.2%), and 79.6% of DRG neurons innervating the periosteum. Co-expression of TRPM7 and TRPV1 among periosteum DRG neurons (75.7%) was more abundant than among cutaneous (53.2%) and muscular (40.4%) DRG neurons. DRG neurons which co-expressed these ion channels in the periosteum had smaller cell bodies compared to the skin and muscle. In addition, the sciatic nerve transection decreased the number of TRPM7-IR neurons in the DRG (approximately 60% reduction). The RT-qPCR analysis also demonstrated reduction of TRPM7 mRNA in the injured DRG. CONCLUSION: The present study suggests that TRPM7 is mainly located in small nociceptors in the DRG. The content of TRPM7 in DRG neurons is probably different among their peripheral targets. TRPM7 in DRG neurons may be able to respond to noxious stimulation from their peripheral tissues. The nerve injury can decrease the level of TRPM7 mRNA and protein in DRG neurons.

Innervation of the human minor salivary glands; immunohistochemical study for neurochemical substances.

Distributions of choline acetyltransferase (ChAT), vasoactive intestinal polypeptide (VIP), dopamine ß-hydroxylase (DBH), tyrosine hydroxylase (TH), neuropeptide Y (NPY) and the transient receptor potential cation channel subfamily V member 2 (TRPV2) were examined in the human minor salivary glands. ChAT-, VIP- and DBH-immunoreactive (-IR) nerve fibers were detected within nerve bundles and close to blood vessels and ducts in the salivary glands. Periacinar nerve fibers were commonly immunoreactive for ChAT in the Ebner's gland but infrequently in other salivary glands. Periacinar VIP-IR nerve fibers were numerous in the palatal gland, moderate in the lingual gland and relatively rare in the labial and Ebner's glands. Some TH-, NPY- and TRPV2-IR nerve fibers were also present around blood vessels and glandular acini in the palatal, lingual and Ebner's glands. Neuronal cells in the vicinity of Ebner's and lingual glands were immunoreactive for ChAT, VIP, TH and TRPV2. By confocal laser scanning microscopy, VIP- and ChAT-IR varicosities were located in the vicinity of myoepithelial and acinar cells in the minor salivary glands. The human minor salivary glands are probably innervated by parasympathetic and sympathetic nerves. Neurotransmitters including neuropeptides in these nerves are thought to be correlated to vasodilation and/or salivary secretion. Acetylcholine and VIP may regulate secretion of the saliva and its components in the salivary glands.

Distribution of neuronal cells which contain dopamine ß-hydroxylase, tyrosine hydroxylase, neuropeptide Y and vasoactive intestinal polypeptide in the human internal carotid nerve.

The human internal carotid nerve (ICN) occasionally has a swelling beneath the external opening of the carotid canal. In this study, the presence and distribution of neuronal cells were investigated in the bilateral ICNs of nine human cadavers. Among 44.4% of the cadavers, swellings were detected in the ICN. Their diameters ranged from 1.7 to 3.6 mm (average ± SD = 2.6 ± 0.7 mm). Thirty-eight percent of these swellings were large (diameter > 3 mm) and showed an oval shape. The large swelling contained many neuronal cells. However, the ICNs with or without a swelling <3 mm diameter were mostly free from neuronal cells (93.3%). Only in one human cadaver, the right ICN without a swelling had a small number of neuronal cells. By the present immunohistochemical method, ICN neurons contained catecholamine-synthesizing enzymes and neuropeptides. Dopamine-beta hydroxylase- and tyrosine hydroxylase-immunoreactivity were mostly expressed by ICN neurons. More than half of them also contained neuropeptide Y-immunoreactivity. However, vasoactive intestinal polypeptide-immunoreactive ICN neurons were relatively infrequent. Substance P- and calcitonin gene-related peptide-immunoreactive ICN neurons could not be detected. By the cell size analysis, neuropeptide Y-immunoreactive neurons were significantly smaller than neuropeptide Y-immunonegative neurons in the ICN. The present study suggests that ICN neurons have a sympathetic function in the human.

Comparison of sputum specimens and nasopharyngeal swab specimens for diagnosis of acute human metapneumovirus-related lower respiratory tract infections in adults.

BACKGROUND: To detect human metapneumovirus, tests besides reverse transcription-polymerase chain reaction (RT-PCR) on nasopharyngeal swab specimens are less accessible. Immunochromatography assays are rapid and simple without the need of any special equipment but sometimes are insufficiently sensitive. This study describes the usefulness of immunochromatography assays to detect human metapneumovirus in adult patients with human metapneumovirus-related acute lower respiratory tract infection using sputum specimens. METHODS: This prospective single-center study enrolled adults and adolescents aged ≥16 years with signs and symptoms of an acute respiratory illness who were diagnosed with acute lower respiratory tract infection. The presence of human metapneumovirus infection was confirmed by seroconversion. Immunochromatography assays and real-time RT-PCR were performed to compare the efficacy of nasopharyngeal swab specimens and sputum specimens. Comparative results were obtained via McNemar's test. RESULTS: Overall, 337 patients were recruited in this study; 63 (18.7%) patients were seroconverted. Sputum specimens showed significantly higher positivity rates than nasopharyngeal swab specimens with both immunochromatography assays (p = 0.0008) and real-time RT-PCR (p = 0.014). Among 29 patients with pneumonia who had concordant positive real-time RT-PCR results for both nasopharyngeal swab specimens and sputum specimens, 21 (72.4%) had a higher viral load in the sputum specimens. CONCLUSIONS: Sputum specimens are more useful in detecting human metapneumovirus than nasopharyngeal swab specimens in adult patients with acute lower respiratory tract infection.