Development of Rat Caries-Induced Pulpitis Model for Vital Pulp Therapy.

Rodent animal models for vital pulp therapy are commonly used in dental research because their tooth anatomy and cellular processes are similar to the anatomy and processes in humans. However, most studies have been conducted using uninfected sound teeth, which makes it difficult to adequately assess the inflammatory shift after vital pulp therapy. In the present study, we aimed to establish a caries-induced pulpitis model based on the conventional rat caries model and then evaluate inflammatory changes during the wound-healing process after pulp capping in a model of reversible pulpitis induced by carious infection. To establish the caries-induced pulpitis model, the pulpal inflammatory status was investigated at different stages of caries progression by immunostaining targeted to specific inflammatory biomarkers. Immunohistochemical staining revealed that both Toll-like receptor 2 and proliferating cell nuclear antigen were expressed in moderate and severe caries-stimulated pulp, indicating that an immune reaction occurred at both stages of caries progression. M2 macrophages were predominant in moderate caries-stimulated pulp, whereas M1 macrophages were predominant in the severe caries-stimulated pulp. Pulp capping in teeth with moderate caries (i.e., teeth with reversible pulpitis) led to complete tertiary dentin formation within 28 d after treatment. Impaired wound healing was observed in teeth with severe caries (i.e., teeth with irreversible pulpitis). During the wound-healing process in reversible pulpitis after pulp capping, M2 macrophages were predominant at all time points; their proliferative capacity was upregulated in the early stage of wound healing compared with healthy pulp. In conclusion, we successfully established a caries-induced pulpitis model for studies of vital pulp therapy. M2 macrophages have an important role in the early stages of the wound-healing process in reversible pulpitis.

Fluorescein transit test time as a tool to assess lacrimal pump function after diode laser transcanalicular dacryocystorhinostomy and external dacryocysto-rhinostomy.

BACKGROUND: Dacryocystorhinostomy (DCR) is the gold standard surgical treatment for nasolacrimal duct obstruction. External DCR is the traditional approach (EXT-DCR); however, the advent of minimally invasive surgeries and the development of optic fiber and laser technologies have made it possible to perform laser transcanalicular DCR (T-DCR), a minimally invasive procedure. This study measured the fluorescein transit time (FTT) after EXT-DCR or T-DCR to evaluate the lacrimal drainage and lacrimal pump function after these two types of DCR. SUBJECTS AND METHODS: A cross-sectional study of 50 patients who underwent EXT-DCR (EXT-DCR Group) or T-DCR (T-DCR Group), who were anatomically patent upon irrigation, with a minimum 6 months of follow up. The patients' FTT was measured; it was defined as the time from the instillation of the dye into conjunctival sac to its free flow from the rhinostomy site. This evaluation was performed through nasal endoscopy performed intranasally with a blue filter that enabled the faster detection of fluorescein from the ostium site. The mean FTTs of the two groups were compared using the two-sided Student's unpaired t-test. Other variables such as sex, age, previous lacrimal sac size, and the site and shape of the rhinostomy were evaluated to determine their possible relationships with FTT. RESULTS: The EXT-DCR group had 80% female patients at a mean age of 58 years. The T-DCR group had the same percentage of female patients (80%) and a mean age of 56 years. The mean FTT group was 47.48 sec in the EXT-DCR and 33.04 sec in the T-DCR group. Functional success was 88% in both groups. CONCLUSION: FTT in the DCR-T Group was significantly lower than in the EXT-DCR Group. No other variables exhibited a statistically significant correlation with FTT. Lacrimal drainage was found to be better after T-DCR than after EXT-DCR, results which show that this procedure could prevent lacrimal pump damage.

Comparative study of Botox injection treatment for upper eyelid retraction with 6-month follow-up in patients with thyroid eye disease in the congestive or fibrotic stage.

AIM: To compare morphometric data of the eyelid fissure and the levator muscle function (LF) before and up to 6 months after transcutaneous injection with five units of Botox in patients with upper lid retraction (ULR) from congestive or fibrotic thyroid eye disease (TED). METHODS: Twenty-four patients with ULR from TED were submitted to transcutaneous injection of 5 units (0.1 ml) of Botox in one eye only. Patients were divided into two groups: 12 with congestive-stage TED (CG), and 12 with fibrotic-stage TED (FG). Bilateral lid fissure measurements using digital imaging and computer-aided analysis were taken at baseline and at regular intervals 2 weeks, 1 month, 3 months and 6 months after unilateral Botox injection. Mean values taken at different follow-up points were compared for the two groups. RESULTS: Most patients experienced marked improvement in ULR, with a mean reduction of 3.81 mm in FG and 3.05 mm in CG. The upper eyelid margin reflex distance, fissure height and total area of exposed interpalpebral fissure were significantly smaller during 1 month in CG and during 3 months in FG. Reduction in LF and in the difference between lateral and medial lid fissure measurements was observed in both groups. The treatment lasted significantly longer in FG than in CG. CONCLUSIONS: A single 5-unit Botox injection improved ULR, reduced LF and produced an adequate lid contour in patients with congestive or fibrotic TED. The effect lasts longer in patients with fibrotic orbitopathy than in patients with congestive orbitopathy.

CD14-mediated induction of interleukin-8 and monocyte chemoattractant protein-1 by a heat-resistant constituent of Porphyromonas gingivalis in endothelial cells.

Viable and inactivated Porphyromonas gingivalis dose-dependently induced interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) secretion in human umbilical vein endothelial cells (HUVECs). The inactivated P. gingivalis, in comparison with viable bacteria, tended to enhance the production of both chemokines more strongly. The production of MCP-1 protein began increasing immediately after stimulation by P. gingivalis, and there was a nearly linear increase from 0 to 8 h of incubation, whereas IL-8 production showed a linear increase between 4 and 12 h of incubation. The IL-8 and MCP-1 mRNA expressions in HUVECs as determined by reverse transcriptase-polymerase chain reaction (RT-PCR) or Quantikine mRNA colorimetric quantification kits were found to be enhanced by P. gingivalis. Furthermore, the time courses of IL-8 and MCP-1 mRNA expressions were in accordance with those of protein production. Addition of polymyxin B or boiling did not weaken the stimulatory effect of P. gingivalis, which inhibited the effect of Escherichia coli lipopolysaccharide (E. coli LPS) and tumour necrosis factor-alpha (TNF-alpha), respectively. In contrast, the induction of IL-8 and MCP-1 by P. gingivalis was significantly reduced by anti-CD14 antibody. Our results suggest that some heat-stable component of P. gingivalis, including LPS, may be responsible for the induction of IL-8 and MCP-1 in HUVECs by a CD14-dependent mechanism. These effects might be involved in the accumulation and activation of neutrophils and monocytes at an early stage of the periodontal pathogenesis.

A heat-stable component of Bartonella henselae upregulates intercellular adhesion molecule-1 expression on vascular endothelial cells.

Bartonella henselae upregulated the expression of intercellular adhesion molecule-1 (ICAM-1) on human umbilical vein endothelial cells (HUVECs). The induction level of ICAM-1 depended on the inoculation bacterial dose. ICAM-1 expression began increasing 4 h after infection and reached a sustained peak beginning at 12 h after B. henselae infection; this time course was similar to that of lipopolysaccharide (LPS) of Escherichia coli. The stimulatory effect was abolished when live B. henselae were separated from HUVECs by a filter membrane. The nonpiliated strain, which is unable to invade endothelial cells, induced ICAM-1 expression to the same extent as the piliated strain. Inactivation of B. henselae by ultraviolet (UV) irradiation, heat (56 degrees C, 30 min), or sonication did not alter its stimulatory activity. Polymyxin B, which strongly inhibited the effect of LPS, did not exert any influence on the stimulatory activity of B. henselae. Furthermore, the effect of sonicated B. henselae was not inhibited even by boiling, which was also the case with LPS. Our data suggest that some heat-stable component of B. henselae binds to the endothelial cell surface, inducing ICAM-1 expression. Though the participation of LPS could not be completely ruled out, we suppose that some unidentified heat-stable proteins, lipids, or polysaccharides may be the stimulatory factor(s). The ability of B. henselae to enhance the expression of adhesion molecules on endothelial cells may be an important mechanism in the pathogenesis of B. henselae infection.

Chronic dacryocystitis in American mucocutaneous leishmaniasis.

PURPOSE: This study describes lacrimal tract involvement and surgical outcome in patients with mucocutaneous leishmaniasis. METHODS: Four patients, ages 20 to 75 years, had nasal lesions resulting from mucocutaneous leishmaniasis and sought treatment for chronic dacryocystitis. Each patient had had lacrimal symptoms since childhood or early adulthood, concomitantly with the development of upper airway lesions. Dacryocystography showed nasolacrimal duct stenosis in all cases on the affected side. Three patients underwent dacryocystorhinostomy (one bilaterally), and one patient had bilateral dacryocystectomy. RESULTS: Two patients had surgical fistula closure soon after surgery. A sequential endoscopic operation for remotion of a synechia between the fistula and the middle turbinate was successful in one of these patients. Histopathologic analysis of lacrimal sacs and nasal mucous membranes close to the anastomotic site revealed chronic nonspecific inflammatory process and negative immunohistochemistry for Leishmania. CONCLUSIONS: Dacryocystitis may result from nasal mucocutaneous leishmaniasis. The surgical outcome was unsatisfactory in one of the four patients.

Live Bartonella henselae enhances endothelial cell proliferation without direct contact.

The proliferation of human umbilical vein endothelial cells (HUVECs) cocultivated with live B. henselae was enhanced in a bacterial dose-dependent manner, and the stimulatory effect was specific to vascular endothelial cells. The inactivation of B. henselae by UV or heat treatment abolished its stimulatory activity, suggesting that live bacteria is necessary for the growth stimulation effect. To investigate the role of direct contact, live B. henselae were separated from HUVECs by a filter membrane (Millicell-CM insert). Even under this condition, an enhanced proliferation of HUVECs was observed. However, no morphological changes in the HUVECs were apparent compared to the B. henselae -infected cells. Furthermore, we isolated a nonpiliated strain of B. henselae that is unable to attach to and enter into endothelial cells. The nonpiliated strain possessed the ability to stimulate the proliferation of cocultivated HUVECs the same as the piliated strain. Moreover, the culture supernatants of B. henselae were also able to induce HUVEC proliferation. Our results indicate that the stimulation of HUVEC proliferation by B. henselae is mediated by soluble factor(s) secreted from the bacteria.

Induced production of nitric oxide and sensitivity of alveolar macrophages derived from mice with different sensitivity to Coxiella burnetii.

We compared in vitro sensitivities to Coxiella burnetii of alveolar macrophages, derived from mice sensitive and resistant to C. burnetii, respectively, and examined the role of nitric oxide (NO) in the C. burnetii infection. Alveolar macrophages of sensitive A/J mice showed a larger population of C. burnetii antigen-positive cells than those of resistant C57BL/6 mice. C. burnetii induced NO production in alveolar macrophages, but N-methyl-L-arginine and sodium nitroprusside (SNP), NO inhibitor and donor, respectively, did not inhibit the infection. Thus the NO induction seems to be independent of the cell defense mechanism against the C. burnetii infection.

[Studies on the locality and the function of spirosome in bacterial cells: relationship between the production of spirosome and anaerobic glycolysis].

The locality and the function of spirosome were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electron microscopy in Peptostreptococcus productus, Clostridium sp. HD-17, Lactobacillus fermentum, Eubacterium aerofaciens and Escherichia cali B. When the bacterial cells in phosphate buffered saline were disrupted by sonication and fractionated by differential centrifugation, the spirosome protein was found in the cytoplasmic fraction and not in the fraction of cell wall and cell membrane (the envelope fraction). The spirosome protein was found, however, in the envelope fraction when the bacterial cells were treated with a proper concentration of SDS. The spirosome protein in the envelope fraction disappeared after treatment with 2% Triton X-100 for 2 h. These results suggest that the spirosome protein anchored to the cell membrane upon SDS treatment. So it is presumed that the locality of spirosome is close to the cell membrane. Spirosome production increased in parallel to the concentration of glucose in the medium in obligate and aerotolerant anaerobes as well as in facultative anaerobes. This result indicates that the spirosome production was induced during the process of anaerobic glycolysis. Fructose as the sole carbon source in the minimal medium induced the spirosome production by E. coli as did glucose, but sodium pyruvate did not induce it either under aerobic or anaerobic condition.

Serological evidence that the Q fever agent (Coxiella burnetii) has spread widely among dairy cattle of Japan.

Serological examination of bovine and human sera for antibodies against Coxiella burnetti was carried out by the immunofluorescence technique. Twenty to 30% of the cows examined were antibody-positive. Sera from two veterinarians also had antibody against C. burnetii. These results suggest an increase in the number of infected cows with C. burnetii in Japan since 1954, and also imply the possibility of the prevalence of acute Q fever in the human population, which had been underestimated and undiagnosed for the last three decades.

Relationship between the production of spirosomes and anaerobic glycolysis activity in Escherichia coli B.

The effects of culture conditions (aerobic or anaerobic) and glucose in the medium on the production of spirosomes in Escherichia coli B were studied by SDS-PAGE and electron microscopy. The Mr of the spirosome of E. coli B was estimated to be 97,000. Electron microscopy revealed that the amount of spirosomes derived from anaerobic cultures was about eightfold larger than that from aerobic cultures. In SDS-PAGE, the bands of spirosome protein derived from anaerobic cultures were more intense than those derived from aerobic cultures, either in peptone water or in Davis-Mingioli's minimal medium. With increased glucose concentration under aerobic conditions, the intensity of the band of spirosome protein was similar to that observed under anaerobic conditions in basal media. These results suggest that spirosome production by E. coli B is related to its anaerobic glycolysis activity.

Observations on the transmission of Angiostrongylus cantonensis from snail to rodent.

A survey of Angiostrongylus cantonensis was carried out to investigate the mode of transmission from mollusc to rat in a fixed study area of Yoron Island from 1979 to 1982. Rattus rattus was found to be infected with a small number of worms in spite of heavy infection with third-stage larvae in Achatina fulica and an abundance of this snail in the area. Natural infection and/or susceptibility with A. cantonensis were confirmed in three small snail species. Bradybaena circulus, Fruticicola despecta and Luchuena reticulata. Young A. fulica was found to be infected with fewer third-stage larvae than mature A. fulica. It was concluded that molluscs which were infected with a small number of third-stage larvae of A. cantonensis play an important role in maintaining the life cycle of A. cantonensis. The percentage of rat stomachs containing mollusc tissue was relatively low, and the incidence and infection was low in rats. Infection with A. cantonensis did not occur very often in R. rattus in nature.

Purification of a kappa-carrageenase from marine Cytophaga species.

A mixture of extracellular carrageenases was isolated from the cell-free medium of a culture of marine Cytophaga sp. 1k-C783 grown on ZoBell 2216 E broth with 0.1% commercial carrageenan. A single active peak of kappa-carrageenase was separated and purified from the mixture by ammonium sulfate precipitation, ion-exchange chromatography, and Sephadex G-200 gel filtration chromatography. Molecular weight of the purified kappa-carrageenase was estimated as 100,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purified kappa-carrageenase had pH optimum 7.6 and temperature optimum 25 C.

Detection of fine spiral structures (spirosomes) by weak sonication in some bacterial strains.

Fine spiral structures (spirosomes) were observed in cell suspensions of five species of bacteria just after weak sonication. The structure is morphologically indistinguishable from the spirosome reported for Lactobacillus species. The molecular weight of the protein of the spirosomes from nine strains was about 94,000 to 95,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The difference in the molecular weight among these spirosomes was not very great, but there were slight differences among the strains from which the spirosomes were derived.

The presence of Angiostrongylus cantonensis in Viti Levu, Fiji.

Wild rats and molluscs were examined for Angiostrongylus cantonensis infection on Viti Levu, Fiji. A. cantonensis were recovered from 29.6% (16/54) of Rattus rattus and 59.5% (25/42) of R. exulans. A. cantonensis-like larval nematodes were found in all of four slugs, Laevicaulis alte, and ten of 20 unidentified land snails. The larvae developed to adult A. cantonensis in the pulmonary arteries of laboratory rats 40 to 42 days after ingestion. This is the first record of A. cantonensis in Fiji.

Differentiation of proactivator from tissue plasminogen activator with low molecular weight in the paranasal mucous membrane.

It has been shown that large amounts of fibrinolytic enzyme are contained in tissue extracts of the paranasal mucous membrane in patients with chronic sinusitis. However, the fibrinolytic enzyme in the tissue extracts has not yet been characterized by biochemical techniques. In the present study, proactivator was differentiated from tissue plasminogen activator with low molecular weight in extracts of the paranasal mucous membrane and the existence of the proactivator was thus demonstrated in tissue extracts. By analogy with the proactivator in antrochoanal polyps, this proactivator may play an important role in the proliferation and enlargement of the paranasal mucous membrane.

[An aetiological study on pustulosis palmaris et plantaris due to tonsillar focal infection (author's transl)]. / Zur Atiologie der Pustulosis palmaris et plantaris, bedingt durch tonsillogene Fokalinfektion.

In clarification of the cause of Pustulosis palmaris et plantaris caused by tonsillar infection 19 patients with this condition and undergoing tonsillectomy were studied. IgE levels in the tonsils and blood serum were estimated both before and after tonsillectomy including various other laboratory tests. These results and the therapeutic effects of the tonsillectomy were analysed. 42 patients with tonsillitis undergoing tonsillectomy were used as controls. Lowered serum complement (C3) titres and decrease in neutrophil counts and serum IgG levels were observed in those patients whose cutaneous lesions were either completely or greatly improved following tonsillectomy. As neither the IgE levels in the tonsils nor in the serum were altered after tonsillectomy, the possibility of IgE mediated allergic type I reaction, as previously proposed by the present authors, was ruled out, and the aetiology was assessed to be an infectious allergy based on the Arthus reaction classified in the type III reaction.

[A study on the preoperative diagnosis of pustulosis palmaris et plantaris due to focal tonsillar infection (author's transl)]. / Zur präoperativen Diagnose der Pustulosis palmaris et plantaris, bedingt durch tonsillogene Fokalinfektion.

In order to establish preoperative tests for Pustulosis palmaris et plantaris due to focal tonsillar infections, a provocation test using ultra-short wave radiation and a negation tests with Impletol solution were investigated. It was subsequently found that the radiation test did not correspond to the therapeutic effects of tonsillectomy on cutaneous lesions, in contrast to the Impletol test. A potentiated dosage of 5 ml Impletol solution was injected submucosally about each palatine tonsil in addition to a conventional 1cc dose (usually injected into the upper pole of each tonsil), and the effects on cutaneous lesions were then examined. In an attempt to define objectively these latter results, both hematologic and serologic tests were obtained, but these failed to show any significant differences in those patients tested.