Sulfate bile acids in germ-free and conventional mice

Sulfated and non-sulfated bile acids were determined in the intestines and in the feces of 7-month-old germ-free and conventional male mice. 1. The bile acid pools in the gall bladder and small intestine were 21.13 mg/100g body weight in germ-free and 11.50 mg in conventional mice. The bile acid pools in the cecum and large intestine of germ-free mice were 3.03 mg/100 g body weight as compared to 1.24 mg in conventional mice. Fecal bile acid excretion was 2.93 mg and 4.12 mg/100 g body weight in 24 h in germ-free and conventional mice respectively. 2. The major bile acids from germ-free mice were cholic acid, alpha-muricholic acid and beta-muricholic acid. Small amounts of chenodeoxycholic and allocholic acid were also present. In addition to these primary bile acids the following secondary bile acids were identified in conventional mice: lithocholic, deoxycholic and omega-muricholic acid. 3. In both germ-free and conventional animals significant amounts of chenodeoxycholic and cholic acid were present as the 7-monosulfate esters. The sulfate esters of these bile acids did not exceed 2% of the total bile acids in the small intestine, but accounted for approximately 50% of the bile acids in the cecum and the large intestine. In contrast, the muricholic acids were nearly exclusively found in the non-sulfate fraction. 4. Alkaline hydrolysis without prior solvolysis of the sulfate esters resulted in loss of bile acids and production of artifacts. Hence, the bile acids of the mouse cannot be analysed by methods involving alkaline deconjugation unless a solvolysis step is included in the procedure.     

Peyer''s-patch-associated synthesis of immunoglobulin in germ-free, specific-pathogen-free, and conventional mice

The synthesis of immunoglobulins G, A, and M has been studied in Peyer's patches together with closely associated intestinal mucosa and in small intestine distant from Peyer's patches in specific-pathogen-free (SPF) Swiss mice and conventional and germ-free C3H mice. Thissue fragments were cultured in vitro in medium containing 14C-labelled amino acids, and newly synthesized proteins were detected by radioimmunoelectrophoresis. Small intestine from SPF and conventional animals synthesized almost exclusively IgA. No immunoglobulin synthesis was detectable in germ-free intestine. In contrast, the Peyer's patches and associated mucosa of all the groups of mice synthesized IgG, IgA, and IgM. This observation is discussed in relation to the possible role of the Peyer's patches as a source of precursors for immunoglobulin-producing cells in the intestine.     

Tooth replantation in germ-free and conventional rats

The purpose of the present study was to evaluate the influence of bacterial infection on the pulpal and periodontal tissues in replanted teeth using germ-free and conventional rats. Forty maxillary and mandibular first molars from ten 6-week-old germ-free male Wistar rats were used. The animals and all materials were maintained in a germ-free environment inside vinyl isolators throughout the experimental periods. Twenty conventional male Wistar rats served as controls. The first molars were intentionally replanted immediately after extraction. At 3 days, and 1, 2, 4 and 8 weeks after replantation, animals were sacrificed and the replanted teeth were histopathologically evaluated. Diversity of pulp tissue response was notable in conventional rats, which initially showed various degrees of neutrophil infiltration and then displayed different types of response, including revascularization with reparative dentin formation and complete necrosis. Pulpal responses of germ-free rats were less variable, being characterized by an almost complete lack of neutrophil infiltration and a high frequency of bone-like tissue ingrowth. Typical inflammatory resorption was detected only in conventional rats, whereas a higher incidence of ankylosis was notable in germ-free rats. The present results may corroborate the concept that bacterial infection is a major cause of serious healing complications following tooth replantation, such as pulp necrosis and inflammatory root resorption. The difficulty in optimally controlling bacterial infection seems to be highly relevant to the complexity and unpredictability of the outcome of this procedure. It should also be emphasized that extensive mechanical damage to the periodontal tissues may trigger the development of unfavorable healing complications as ankylosis, even under strictly aseptic conditions.     

Ex vivo gastrointestinal biotransformation of chlorothalonil in the germ-free and conventional rat

1. The metabolism and absorption of chlorothalonil and corresponding diglutathione and dicysteine conjugates was studied using isolated everted gastrointestinal sacs of the conventional and germ-free rat. An HPLC method was used to analyse mucosal and serosal fluids. Thiol metabolites of chlorothalonil were determined by GC/MS. 2. Low absorption of the substrates was observed, with < 4% of the radioactivity being recovered from the serosal buffers and the digestive tissues. A major part of the radioactivity was recovered from the mucosal fluids and it corresponded to unchanged chlorothalonil. Traces of unchanged chlorothalonil and mono-, di- and trimethylthio metabolites were present in serosal fluids as well as unidentified polar peaks. An important transformation (> 75%) of the chlorothalonil conjugates was observed. The di- and trimethylthio metabolites of chlorothalonil were detected from both sides of the everted gut sac of rat incubated with the diglutathione and dicysteine conjugates. 3. Few differences were observed between the conventional and germ-free rat: absorption was higher in the duodenum of germ-free rat, but tissue retention was more significant in the duodenum of the conventional rat.     

感染根管内细菌定植状态的观察

目的:了解慢性根尖周炎患牙根管内的细菌感染状态,以指导临床治疗和评估预后.方珐:采用组织学与扫描电镜相结合的方法对临床不能保留的7例慢性根尖周炎患根进行超微结构观察,将新鲜拔除的患根沿牙长轴近、远、中向劈为两半,一半做组织学切片,采用Brown&Brenn染色观察,另一半做扫描电镜观察.结果:组织学切片细菌特染显示,大量细菌附着于根管壁表面并定植于根管壁牙本质小管深部,用测微尺测量细菌侵入深度为140～1000μm,根尖1/3段感染较重.扫描电镜结果显示,根管壁表面可见到致密、半封闭状态的网状细菌膜结构,大量球菌、杆菌和/或丝状菌镶嵌于无定形的基质中,集中于根尖1/3段.结论:慢性根尖周炎的根管内细菌以生物膜形式定植在根尖1/3区,并侵入牙本质小管深部,形成感染难以清除的区域.     

The challenge of preparing the curved root canal

Heavily restored teeth which become pulpally involved are now often root canal treated rather than extracted. While this is laudable it has a significant impact on the practice of endodontics today. The curved calcified canal can prove very difficult to prepare to its natural shape by conventional techniques and there is always the likelihood of iatrogenic damage. To compensate for this many clinicians tend to under-prepare these canals, perhaps leaving them inadequately cleaned and certainly remarkably difficult to fill to length. The purpose of this article is to outline the stages of and the rationale behind a hand instrumentation preparation method which uses the balanced force method of movement of files from canal entrance to apical constriction. In the authors' experience this technique has gone a long way towards solving the problem of cleaning and shaping the fine curved canal. With some practice, but no extra expense, the technique described will not only speed up canal preparation but will also make it more predictable.     

Experimental feline panleucopenia in the conventional cat

Coventional kittens, 12-27 weeks old, were inoculated with cell-cultured feline panleucopenia virus and killed sequentially between day 3 and day 24 after inoculation. All developed a non-fatal mild disease between days 2 and 9, characterized by lymphopenia, neutropenia, listlessness, depression and the development of neutralizing antibodies to the virus. Small intestinal bacterial counts were reduced during the period of maximal clinical disease, presumably a result of decreased food intake. There was involution of the thymus with marked depletion of lymphocytes between days 3 and 12. Depletion of lymphocytes also characterized the lesions in the lymph nodes between days 3 and 8. At the same time crypt lesions with spotty distribution were in the small intestinal and colonic mucosa. The changes were loss of crypt epithelial cells with compensatory attenuation of the remaining epithelium. Electron microscopically, the number and size of microvilli and secretory granules were reduced but there was no change indicating lethal cell injury. There were no virus particles. The findings point to an early and transient cellular damage by the virus. Intestinal alkaline phosphatase activity disappeared from the luminal surface of the attenuated crypt epithelial cells. Otherwise, intestinal alkaline and acid phosphatase activity were not altered in inoculated cats.     

Carbon fiber reinforced root canal posts. Mechanical and cytotoxic properties

The aim of this study was to compare the mechanical properties of a prefabricated root canal post made of carbon fiber reinforced composites (CFRC) with metal posts and to assess the cytotoxic effects elicited. Flexural modulus and ultimate flexural strength was determined by 3 point loading after CRFC posts had been stored either dry or in water. The bending test was carried out with and without preceding thermocycling of the CFRC posts. The cytotoxicity was evaluated by an agar overlay method after dry and wet storage. The values of flexural modulus and ultimate flexural strength were for dry stored CFRC post 82 +/- 6 GPa and 1154 +/- 65 MPa respectively. The flexural values decreased significantly after water storage and after thermocycling. No cytotoxic effects were observed adjacent to any CFRC post. Although fiber reinforced composites may have the potential to replace metals in many clinical situations, additional research is needed to ensure a satisfying life-span.     

Thermographic assessment of root canal obturation using thermomechanical compaction

The potential interaction is described between caffeine and antipyrine, two drugs with a high probability of being concomitantly administered for the evaluation of liver metabolism. In order to determine the influence of antipyrine on the elimination of caffeine, salivary caffeine clearance was measured in six healthy volunteers prior to and 2 and 5 days after the administration of a single oral dose of 1000 mg of antipyrine. Total caffeine clearance increased on average by 24% (from 1.65 to 2.05 ml/min, P = 0.1) 2 days after antipyrine dosing, and 25% (from 1.65 to 2.06 ml/min, P < 0.01) 5 days after the administration of antipyrine, whereas the half-life decreased by around 24% (from 5.3 to 4 h, P = 0.09) after 2 days and 26% (from 5.3 to 3.9 h, P = 0.05) after 5 days. The apparent volume of distribution did not change. These results suggest that antipyrine is able to increase the elimination of caffeine, probably by means of inducing its hepatic metabolism. When both drugs are used sequentially in the same patient to assess the drug metabolizing activity of the liver, the caffeine test should be performed first.     

Experimental study of the biocompatibility of four root canal sealers and their influence on the zinc and calcium content of several tissues

Four root canal sealers (AH-26, Roth 811, CRCS, and Sealapex) were tested for tissue biocompatibility in rat connective tissue. Each sealer was placed in Teflon tubes and implanted subcutaneously in Wistar-Furth rats. The implants were removed after 7, 14, and 21 days, fixed, and histologically prepared for microscopical evaluation. Brain, liver, kidneys, and uterus were removed from the animals killed at the first experimental period (7 days) and analyzed for zinc and calcium concentration by flame atomic absorption spectrophotometry. In total, 100 specimens were examined. At the seventh day, the most irritant material was seen to be AH-26, but this inflammatory reaction decreased with time. Roth 811 and Sealapex caused moderate-to-severe inflammatory reaction, whereas CRCS caused mild to moderate. CRCS and Roth 811 induced redistribution of zinc, whereas AH-26 induced changes in calcium content in some organs.     

Changes in cell proliferative activity in granulation tissue invading the root canal

Root canal models were implanted in rats in order to investigate histologically the movement of granulation tissue invading apical canals and dead spaces and changes in cell proliferative activity as indicated by 3H-thymidine. The models were prepared to have 1, 2, and 3 mm apical canals, with and without dead spaces, to simulate preparations short of the apex and obturation to several levels. After 12 wks implantation of the models without dead spaces, granulation tissue invading 1 mm apical canals did not degenerate and cell proliferative activity remained high. However, tissue invading the 2 and 3 mm apical canals tended to be necrotic and cell proliferative activity was decreased. In the models with dead spaces, the tissue in the 1 and 2 mm apical canals developed and invaded the dead spaces, and still possessed proliferative activity 12 wks after implantation. In contrast, the tissue in the 3 mm apical canals did not invade the dead spaces, even after 12 wks, and no proliferative activity was observed.     

pH changes in root dentin over a 4-week period following root canal dressing with calcium hydroxide

An immunofluorometric method was developed for the semiquantitative determination of trihexosylceramide in cultured fibroblasts from Fabry disease patients, using a laser scanning confocal imaging system. The accumulated glycolipid was detected as granular inclusions in the cells. Heterozygote identification was achieved both by counting of immunoreactive cells and by measuring the relative fluorescence intensity with a digital imaging system.     

Initial biological testing of root canal sealing materials--a critical review

A number of systems for the biological testing of dental materials have been suggested. This paper presents a critical review of the protocols suggested in the ISO standards for the initial and secondary biological testing of root canal sealing materials.     

Follow up after root canal treatment of young permanent molars

While the majority of experimental cervical spine biomechanics research has been conducted using slowly applied forces and/or moments, or dynamically applied forces with contact, little research has been performed to delineate the biomechanics of the human neck under inertial "noncontact" type forces. This study was designed to develop a comprehensive methodology to induce these loads. A minisled pendulum experimental setup was designed to test specimens (such as human cadaver neck) at subfailure or failure levels under different loading modalities including flexion, extension, and lateral bending. The system allows acceleration/deceleration input with varying wave form shapes. The test setup dynamically records the input and output strength information such as forces, accelerations, moments, and angular velocities; it also has the flexibility to obtain the temporal overall and local kinematic data of the cervical spine components at every vertebral level. These data will permit a complete biomechanical structural analysis. In this paper, the feasibility of the methodology is demonstrated by subjecting a human cadaver head-neck complex with intact musculature and skin under inertial flexion and extension whiplash loading at two velocities.     

Derivation of a breeding colony of germ-free athymic mice by cesarean section and foster nursing

Germ-free athymic mice were obtained by removing the uterus by cesarean section from a gravid heterozygous (nu/+) female at precisely full term. The uterus was transferred into a germ-free flexible isolator containing two germ-free CD-1 mice that were communally nursing their recently delivered litters. These germ-free CD-1 mice served as foster mothers for the newborn germ-free athymic mice.