Expression of fibroblast growth factor receptor1, -2c, and -3c transcripts in mouse molars after tooth eruption.

A previous study suggested that fibroblast growth factor (FGF) signaling plays an important role in dentin formation during tooth development. In this study, to examine dentin formation after tooth eruption involving secondary and tertiary dentin, we analyzed the expression patterns and expressing cells of Fgfr1, -2c, and -3c in mouse maxillary first molars (M1). Since it is difficult to recover the mRNAs from mineralized tissues, we tested methods for extraction after fixation and decalcification of teeth. We successfully obtained consistent results with quantitative real-time PCR (qPCR) using ß-actin transcripts for validation. qPCR for Dentin sialo phosphoprotein (Dspp), Fgfr1, -2c, and -3c transcripts was performed on mice at ages of 2-20 weeks. The results showed that the highest expression levels of Dspp and Fgfr2c occurred at 2 weeks old followed by lower expression levels after 4 weeks old. However, the expression levels of Fgfr1 and Fgfr3c were constant throughout the experimental period. By in situ hybridization, Dspp, Fgfr1, and Fgfr3c transcripts were detected in odontoblasts at ages of 2 and 4 weeks. In addition, Dspp and Fgfr1 transcripts were detected in odontoblasts facing reactionary dentin at 8 weeks old. These results suggest that FGF-FGFR signaling might be involved in the regulation of odontoblasts even after tooth eruption, including secondary and tertiary dentin formation. Moreover, our modified method for extracting mRNA from mineralized tissues after fixation and decalcification successfully produced consistent results.

Anatomical status of the human palatopharyngeal sphincter and its functional implications.

PURPOSE: The transition muscle between the palatopharyngeus (PP) and the superior constrictor of the pharynx (SCP) encircles the pharyngeal isthmus from behind and is designated as the palatopharyngeal sphincter (PPS). The PPS is inferred to play important roles for velopharyngeal closure, but its existence remains controversial and its roles have been regarded as being played by the SCP. The present study aimed to clarify the anatomical status and functional implications of the PPS. MATERIALS AND METHODS: Macroscopic and microscopic examinations were performed on 39 and 4 cadavers, respectively. In the former, the bilateral PPSs and their adjacent structures were exposed from outside and/or inside. In the latter, the velums embedded in paraffin were cut into frontal or sagittal sections and alternately processed with HE and Azan stains. RESULTS: The PPS originated from the nasal aspect of the lateral half of the palatine aponeurosis and the inferior margin of the medial pterygoid plate and was distinguishable from the PP descending in and along the palatopharyngeal arch and the cranialmost portion of the SCP in its origin. It passed dorsally on the lateral side of the levator veli palatini and traversed around the salpingopharyngeal fold running longitudinally. It then entered below the SCP and ran toward the pharyngeal raphe with SCP muscle fibers intermingled. CONCLUSIONS: The PPS is a muscle distinct from the SCP. Its contraction produces Passavant's ridge and conceivably enhances the efficiency of velopharyngeal closure by pressing the salpingopharyngeal fold and the musculus uvulae ridge against the velum.

Gross anatomical classification of the courses of the human lingual artery.

PURPOSE: There are many reports on the variation of origin site of the lingual artery branching from the external carotid artery. However, there are few reports systematically investigating the course of the lingual artery in detail from branching site to the body of tongue. The purpose of this study is to classify systematically the courses of the lingual artery including variations. METHODS: Using 111 body sides of 63 Japanese cadavers for dissection practices, the lingual artery and the surrounding structures were investigated gross anatomically. RESULTS: The courses of the lingual artery were classified into five types based on the positional relationships with the hyoglossus and the mylohyoid as follows; type M: coursing medial to the hyoglossus (normal course, 104 sides), type L: coursing lateral to the hyoglossus (2 sides), type T: transferring its course from lateral to medial to the hyoglossus (2 sides), type P: penetrating the mylohyoid (2 sides), and type C: the coinciding of types M and P (1 side). Types L, T, P, and C were considered to be variant lingual arteries. Types M and T, type L, and type P arose from the external carotid, facial, and submental arteries, respectively. In types L and P, adding to the variant lingual artery, the remnant of the normal lingual artery was also observed. CONCLUSION: The present study provides detailed information on the courses of lingual artery which will be of clinical importance especially in the super-selective arterial angiography.

Gross anatomical classification of the courses of the human sublingual artery.

The purpose of the present study was to classify the courses of the human sublingual artery. For this purpose, the arteries supplying the floor of the mouth and the tongue were gross anatomically investigated using 101 sides of 53 cadavers. The courses were divided into three categories: those passing medial or lateral to the hyoglossus (categories M and L) and that piercing the mylohyoid (category P); they were subdivided into five types. Category M had one type, regarded as the usual one, in which the lingual artery took the usual pattern of distribution. Categories L and P, in which the sublingual artery arose from the facial or submental artery, had the respective two types and were collectively regarded as the unusual type. Sixty-one and 36 of the 101 sides were of the usual and unusual types, respectively, the latter of which included 17 of category L and 19 of category P. The remaining four were variations of the lingual artery itself. On examining the types by gender, the usual type was more often found in females (75.6%), whereas the unusual type was more often found in males (48.1%). Bilateral occurrence of the same type was often found in both the usual type (77.4%) and the unusual type (65.0%). Existence of the sublingual artery branch significantly increased the thicknesses of the submental arteries. The classification proposed here will conceivably contribute to safer dental implant surgery and more accurate interpretation of angiographic images of arteries in the floor of the mouth.

Anatomical status of the human musculus uvulae and its functional implications.

In our ongoing series of anatomical studies to determine the three-dimensional architecture of the human velar muscles, we have previously reported on the palatopharyngeus. The present study deals with the musculus uvulae (MU), in which the positional relationships of its origin to the posterior nasal spine and the palatine aponeurosis, as well as the interrelation between its anatomical status and functions, have yet to be clarified. Macroscopic and microscopic examinations were performed on 25 and 2 cadavers, respectively. In the former, bilateral MUs and their adjacent structures were exposed mainly from the nasal aspect. In the latter, the soft palates embedded in paraffin were cut into frontal and sagittal sections and alternately processed with HE and Azan stains. The left and right MUs adjacent to each other were found to run longitudinally along the midline beneath the nasal aspect of velum. It was overlaid by glandular tissue that increased in amount as it coursed distally. After originating from the oral surface of palatine aponeurosis, it ran backward to cross above the sling formed by the levator veli palatini muscles of both sides and reached the tip of uvula with its muscle fibers intermingled with glandular tissue. Past studies have proposed three functions of MU to enhance the efficiency of velopharyngeal closure: space occupier, stiffness modifier, and velar extensor. All of the above-described anatomical characteristics of MU could be explained as being adapted for these functions. This implies that MU is actively responsible for maintaining the velopharyngeal closure efficiency.

Gross anatomical study of the human palatopharyngeus muscle throughout its entire course from origin to insertion.

The palatopharyngeus (PP) extends throughout the entire length of the pharynx and probably plays an important role in deglutition, but its spatial distribution remains undefined in some respects. This study was designed to clarify the exact distribution of the PP indispensable for understanding its functions. Using 50 cadavers, the PP and its neighboring muscles were bilaterally exposed in both surfaces of the pharynx. The PP was composed of two divisions: longitudinal and transverse. It is already known that the longitudinal PP is divided into two fasciculi sandwiching the levator veli palatini (LVP) immediately after originating from the palatine aponeurosis. However, we newly discovered a fasciculus originating from the uvula, and further regarded the salpingopharyngeus as another fasciculus of origin. The four fasciculi united to descend through the palatopharyngeal arch and inserted into the thyroid cartilage and beneath the mucosa of the hypopharynx. The transverse PP occupied a location transitional between the PP and superior constrictor (SC), where it originated from the palatine aponeurosis and passed dorsally to encircle the pharyngeal isthmus and reach the pharyngeal raphe. Although whether it belongs to the PP or SC has remained controversial, we regarded it as a portion of the PP from the evolutionary perspective and proposed anatomical criteria for differentiating it from the SC. The wide distribution of the PP suggests that it acts not only to elevate the pharynx or depress the soft palate, but also as a nasopharyngeal sphincter when closing the pharyngeal isthmus.

Influenza vaccine with Surfacten, a modified pulmonary surfactant, induces systemic and mucosal immune responses without side effects in minipigs.

Immune responses and side effects of intranasally administered flu vaccine with the commercial product Surfacten, a modified bovine pulmonary surfactant, were investigated in minipigs. The use of minipigs was based on the anatomical resemblance of nasal lymph nodes, the principal antigen uptake site of respiratory mucosal immunity, between pig and human. Intranasal instillation of HA vaccine adjuvanted with Surfacten elicited significantly higher serum hemagglutination inhibition titers than the antigen alone, with wide cross-neutralizing activities of secretory IgA in nasal washes. No significant induction of inflammatory cytokines or migration of inflammatory cells was observed at the site of immunization or serum after the first immunization. These data suggest the potential usefulness of Surfacten for mucosal vaccination.

The effects inhibiting the proliferation of cancer cells by far-infrared radiation (FIR) are controlled by the basal expression level of heat shock protein (HSP) 70A.

We developed a tissue culture incubator that can continuously irradiate cells with far-infrared radiation (FIR) of wavelengths between 4 and 20 microm with a peak of 7-12 microm, and found that FIR caused different inhibiting effects to five human cancer cell lines, namely A431 (vulva), HSC3 (tongue), Sa3 (gingiva), A549 (lung), and MCF7 (breast). Then, in order to make clear the control system for the effect of FIR, the gene expression concerned to the inhibition effect by FIR were analyzed. In consequence, basal expression level of HSP70A mRNA was higher in A431 and MCF7 cells than in the FIR-sensitive HSC3, Sa3, and A549 cells. Also, the over expression of HSP70 inhibited FIR-induced growth arrest in HSC3 cells, and an HSP70 siRNA inhibited the proliferation of A431 cells by irradiation with FIR. These results indicate that the effect of a body temperature range of FIR suppressing the proliferation of some cancer cells is controlled by the basal expression level of heat shock protein (HSP) 70A. This finding suggested that FIR should be very effective medical treatment for some cancer cells which have a low level of HSP70. Still more, if the level of HSP70 in any cancer of a patient was measured, the effect of medical treatment by FIR can be foreseen for the cancer.