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PubMed1986

Human gingival attachment responses to subgingival crown placement. Marginal remodelling.

13 teeth in block were extracted from 2 patients. Their facial periodontal condition was essentially within normal clinical limits. Temporary crowns covering the bevel were placed below the base of the crevice 1 to 8 weeks prior to extraction. At time of extraction, all blocks were decalcified, the temporary crown dissolved, and the blocks prepared for histologic examinations using buccolingual cut, step serial sections. Histologic data revealed reformation of a new supracrestal attachment unit within 1 week following crown placement. The reformation of the gingival unit consisted of marginal recession with apical and lateral migration of the junctional epithelium to the level of remaining cementum inserted fibers. With gingival recession and migration of junctional epithelium, resorption of crestal portions of the facial plate occurred. However, periodontal fibers anchored into cementum opposite the resorbed bone were not lysed. Rather, the attached fibrillar ends appeared to interdigitate with fibers from the corium of the facial gingiva at this site, thereby forming a more apically located crestal attachment. This response may be one mechanism of reformation of the gingival attachment unit taking place following mechanical and/or surgical injury to this site and is completed often, within 2 weeks after injury.

Connective Tissue↗
PubMed2003

Specific detection of human BK polyomavirus in urine samples of immunocompromised patients.

A semiquantitative PCR assay for the detection of BK virus in urine was developed using primers for BK virus that specifically amplified BK but not JC virus. DNA was extracted from urine through treatment with proteinase K followed by DNA precipitation with sodium acetate. Semiquantitation was achieved by amplifying serial dilutions (1:1, 1:10, 1:100, and 1:1,000) of the urine specimens. Each assay included both positive (stock BK virus and previously positive patient urine) and negative (no template) controls. A urine sample was interpreted as positive if any of the serial dilutions showed amplification of the DNA fragment of the expected size. For some patient-derived samples, amplification of the expected-size fragment was achieved with a dilute template whereas no amplification was achieved with a concentrated template. This was attributed to interfering substances in the urine. PCR results were compared with urine cytology and shown to be more sensitive. Validation studies were performed at the University of Nebraska Medical Center, utilizing a separate qualitative PCR assay that detects both BK and JC virus and distinguishes between them by restriction enzyme digestion patterns. Of 46 urine samples analyzed using both methods, 22 were positive by both assays, 18 were negative by both assays, 5 were positive only by the Nebraska method, and 1 was positive only by our method. In comparison with the Nebraska PCR, our PCR assay had a sensitivity of 81% and specificity of 95%. For twenty-one (43%) of 49 immunocompromised patients, tests were positive when specimens were submitted because of clinical suspicion of BK virus infection.

BK Virus↗
PubMed1989

Inhibition of human neutrophils by auranofin: chemotaxis and metabolism of arachidonate via the 5-lipoxygenase pathway.

The effect of auranofin on human neutrophil (PMN) 5-lipoxygenase activity and leucotriene B4 (LTB4) chemotaxis was investigated. [1-14C]Arachidonic acid was incorporated into the purified cells until steady state conditions were obtained. After preincubations with serial dilutions of auranofin arachidonic acid release and metabolism were stimulated with calcium ionophore A23187. The radioactive eicosanoids released were extracted and separated by thin layer chromatography, followed by autoradiography and quantitative laser densitometry. Chemotaxis of PMNs towards LTB4 was measured in a modified Boyden chamber. Auranofin showed dose dependent inhibition of both the 5-lipoxygenase pathway (IC50 17.4 X 10(-6) mol/l) and of chemotaxis (IC50 45 X 10(-6) mol/l). The release of arachidonic acid from phospholipids was unaffected in the concentration range tested (1-1000 mumol/l). Inhibition of both neutrophil motility and cellular synthesis of proinflammatory eicosanoids may thus contribute to the beneficial clinical effects of auranofin in rheumatoid arthritis.

Arachidonate 5-Lipoxygenase↗
PubMed1975

Localization of luteinizing hormone-releasing hormone in the preoptic area and hypothalamus of the rat using radioimmunoassay.

To determine the localization of luteinizing hormone-releasing hormone (LHRH), five brains from adult male rats were serially sectioned in a cryostat at - 10 C in either the frontal, horizontal or sagittal planes. Acetic acid-ethanol extracts of each section were assayed for LHRH using radioimmunoassay (RIA) and in some cases using bioassay as well. Approximately 0.2 ng LHRH was concentrated in medial basal preoptic (MB-PO) tissue overlying the rostral portion of the optic chiasm. This LHRH appears to be associated with the organum vasculosum of the lamina terminalis and/or adjacent neural tissue. Uniform, low levels of LHRH were detected in hypothalamic tissue between the preoptic area (POA) and arcuate-median eminence (ARC-ME) region. In the ARC-ME region 2.7 ng of LHRH were concentrated primarily in the median eminence. The lateral distribution of LHRH in the ARC-ME region extended beyond the median eminence into tissue corresponding to the lateral aspect of the ventromedial nucleus. Concomitant bioassay and RIA determinations of LHRH were highly correlated. Of the sections bioassayed, only those sections containing LHRH released FSH. These results confirm the presence of LHRH in the POA and in the rostral hypothalamus of the rat brain. The possible significance of LHRH in the POA for the regulation of LH release is discussed.

Animals↗
PubMed1975

The effect of hypoglycemia on hypothalamic thyrotropin-releasing hormone (HRH) in the rat.

A rapid radioimmunoassay for TRH with a high degree of specificity and sensitivity is described. The procedure is capable of measuring TRH in amounts less than 15 pg/tube. Such an assay system has enabled us to study the effect of hypoglycemia on hypothalamic TRH content. Forty-eight female, adult Holtzman rats were divided into 8 groups of 6. Regular insulin was injected intraperitoneally into each rat except for the basal group. A separate batch of similar rats was studied in the same fashion except that saline was injected instead of insulin. Each group of rats was decapitated and the trunk blood collected at 0, 15, 30, 45, 60, 90, 120 and 180 min post-injection. Appropriate tissues were rapidly taken and immediately extracted in ice-cold methanol. Hypothalamic TRH, pituitary TSH, serum TSH and serum triiodothyronine (T3) and thyroxine (T4) were serially determined. In the insulin-treated group, a rapid fall in blood sugar was observed reaching a nadir in 15 min. Hypothalamic TRH fell from a basal mean +/- SE value of 3.25 +/- 0.31 ng to 1.54 +/- 0.14 ng/hypothalamus (P less than 0.01). Pituitary TSH decreased from 10.0 +/- 0.9 mug to a low of 2.6 +/- 0,8 mug/pituitary (P less than 0.02) at 30 min postinsulin. Serum TSH increased from a basal level of 42.5 +/- 20.5 muU/mo to a peak of 102.1 +/- 10.0 muU/ml 45 min (P less than 0.05) after insulin administration. The incremental change in serum T3 occurred at 90 min when T3 levels increased from a baseline of 107.5 +/- 53.7 ng/100 ml to a peak of 711.7 +/- 20.2 ng/100 ml (P less than 0.01). No changes in T4 were observed. The control group of rats did not show significant changes in hypothalamic TRH. The results of the study indicate that hypoglycemia can induce depletion (presumably release) of hypothalamic TRH with a consequent cascade stimulation of the pituitary-thyroid axis.

Animals↗
PubMedDate not supplied

[Implantation of flakes of castor oil resin in rat dental alveolus].

The purposes of the present study were: 1) to investigate the biocompatibility of a natural resin (made of fatty acids extracted from Ricinus communis) implanted in the dental alveolus of rats and 2) to verify any possible interference of that material in the osseous healing following tooth extraction. The resin (AUG-EX, Poliquil Araraquara Polímeros Químicos LTDA, Araraquara--SP) was placed inside de alveoli immediately after extraction of the upper right incisors. The animals were sacrificed 1, 2, 3 and 6 weeks after extraction or extraction + implantation. The hemi-maxillae were decalcified and processed for paraffin embedding. Longitudinal 6-micrometer-thick semi-serial sections stained with hematoxylin and eosin were obtained. Histologic examination showed particles of irregular shape and variable size (700-1200 microns) localized in the medium/cervical alveolar thirds, with a scanty but persistent foreign body reaction. From the second week on, as the relative volume of bone trabeculae increased, it was seen in close contact with the surface of the implanted material in some regions. Histometric analysis (differential point counting method), used to quantify the healing process in the apical third, showed a small but significant decrease (13%-20%) in new bone formation in the implanted rats. In conclusion, the results show that, in spite of its biocompatible nature, the studied resin hinders the post-extration healing process.

Animals↗
PubMed2001

Focal testicular lesion after sperm extraction or aspiration: sonographic appearance simulating testicular tumor.

OBJECTIVE. We describe the sonographic features of focal intratesticular lesions seen in men who underwent sperm retrieval procedures. CONCLUSION. Although many urologists believe that solid intratesticular masses are malignant until proven otherwise, a growing number of benign focal testicular lesions have been described. Awareness of the cause and sonographic appearance of focal abnormalities in men who have undergone testicular aspiration or extraction should help radiologists suggest the correct diagnosis and advise a conservative approach on the basis of close surveillance by serial physical, laboratory, and imaging studies.

Adult↗
PubMed2005

Bone formation in beta-tricalcium phosphate-filled bone defects of the rat femur: morphometric analysis and expression of bone related protein mRNA.

The purpose of the current study was to evaluate the bone formation when beta-tricalcium phosphate (TCP) was implanted in bone defects of rat femurs. beta-TCP granules were applied to defects created in the femurs of 65 male rats who were sacrificed 3, 7, 10, 14 or 30 days later. Bone tissues were embedded in paraffin, serial sections were cut and then stained with hematoxylin-eosin. Histomorphometric analyses were also conducted. Furthermore, total mRNAs were extracted, homogenized, and reverse transcribed, after which quantitative PCR assays were conducted with a LightCycler using the double-stranded DNA dye Syber Green I with primers for either rat osteopontin or osteocalcin. Tissues in defects without beta-TCP were used as controls. The amount of newly formed bone tissue in the beta-TCP implanted group was significantly greater in both the side areas and the central area of defects than in the control group. Expressions of osteopontin and osteocalcin mRNAs of cells in the defects of the experimental group were up-regulated compared with the control group at all time periods. Taken together, these results prove that beta-TCP is an appropriate material for osteoconduction and promotes bone formation in bone defects.

Animals↗
PubMed1992

[Histological study of the change of pulp tissue during shedding of the deciduous tooth].

To understand the pulp change of root resorption of human deciduous teeth from a histological point of view, one hundred and twenty specimens were collected for study. The teeth (caries-free) were extracted for occlusal reason or because they exhibited various degrees of mobility and discomfort. The extent of root resorption was classified into five grades: Res. i, Res. 1/4, Res. 1/2, Res. 3/4 and Res. Co., according to Moorrees et al. and Haavikko. All teeth were fixed in 10% formalin (pH 7.2) immediately after extraction for 3-7 days. They were then decalcified for 14 days in 5% formic acid-sodium citrate, sectioned serially at 7 microns and stained with hematoxylin and eosin, Masson's stain, and silver impregnated stain. All sections were examined under a light microscope. The results obtained were as follows: 1. While the root was resorbed more than one half way, some normal pulp was replaced by the connective tissue as in inflammation. Internal resorption could be seen. 2. By the time deciduous root resorption was essential as complete, normal pulp tissue was no longer present. 3. Because the nerve degenerated during initial tooth resorption, there was no evidence that deciduous root resorption was under nervous control. 4. The present observation suggests that in addition to odontoclasts, several other cells such as fibroblast-like cells and macrophages, were actively involved in root resorption during the shedding of deciduous teeth.

Child↗
PubMed2004

[The effect of Redix Scutellariae on butyrate of Porphyromonas endodontalis in vitro].

OBJECTIVE: To study the effect of Radix Scutellariae on the growth, metabolism of Porphyromonas endodontalis (P.e), as a preparation for studying the mechanism of Radix Scutellariae in treating pulp and periapical diseases. METHODS: P.e was chosen as the experimental bacteria. Radix Scutellariae was extracted by means of reflux with 80% ethanol. The value of MIC of Radix Scutellariae was measured by minute amount serial dilusion test, and the production of butyrate was measured by high liquid chromatograph(HPLC). RESULTS: Radix Scutellariae could inhibit the growth of P.e, of which the MIC was 100 mg/L. Following the increase in concentration of Radix Scutellariae, the amount of butyrate decreased to (3.527 +/- 0.009) mg/L, (3.048 +/- 0.005) mg/L, (2.490 +/- 0.011) mg/L, (2.209 +/- 0.016) mg/L, respectively (P < 0.05). CONCLUSION: Radix Scutellariae could inhibit the growth and metabolism of P.e and might be an effective agent in treating pulp and periapical diseases.

Bacteroidaceae Infections↗
PubMed1991

Kinetic ELISA for detection of antibodies to Psoroptes sp. (Acari: Psoroptidae) in bighorn sheep (Ovis canadensis).

A kinetic enzyme-linked immunosorbent assay (ELISA) was developed using antigenic extracts prepared from Psoroptes cuniculi mites and sera from 37 Psoroptes sp.-infested and 43 uninfested bighorn sheep (Ovis canadensis). Serial dilutions of these serum samples, representing 3 bighorn sheep subspecies and 9 geographic areas, gave parallel responses when plotted as log dilution versus log kinetic rate. Therefore, all 80 samples were run at a single dilution (1:100) and positive/negative cutoff values were established as the mean kinetic rate of all negative sera plus either 2, 3, or 4 standard deviations. The resulting ELISA was highly reproducible and accurate with sensitivities and specificities of 100% and 97.7%, 94.6% and 97.7%, and 94.6% and 100%, respectively. This immunoassay will be useful for prospective and retrospective studies assessing the distribution and prevalence of Psoroptes sp. infestations in bighorn sheep.

Animals↗
PubMed1990

Effects of various irrigating solutions on the cleaning of the root canal with ultrasonic instrumentation.

The authors studied, in vitro, by light microscopy and by morphometric analysis, the cleaning capacity of Dakin's solution, water and Tergentol, when used as auxiliary solutions to ultrasonic instrumentation of root canals. After irrigation and instrumentation, 15 extracted human mandibular incisors, with a single root and with only one canal, were submitted to routine histological processing with serial 6-microns sections stained with hematoxylin and eosin. The results showed that Dakin's solution energized by ultrasound leaves root canals with less debris than Tergentol; water falls into an intermediate position. The apical third showed more debris than the middle third, and none of the irrigating solutions left the root canals free of debris.

Bicarbonates↗
PubMed1984

A standardized method of detecting antibodies to extractable nuclear antigens (RNP and Sm) by gel precipitation.

Because of the importance of the detection of antibodies to RNP and Sm in the diagnosis of mixed connective tissue disease, systemic lupus erythematosus and certain forms of systemic sclerosis, the various factors which influence the sensitivity of the gel precipitation method for the detection of these antibodies were investigated. The agarose concentration, thickness, well sizes and distance between wells influence the sensitivity of the precipitin reactions. Suitable conditions for a sensitive and reproducible test system are specified. Because of variations in the antigenic composition of extractable nuclear antigen preparations made by different methods and from different tissue sources, a two-dimensional "chessboard titration" using serial twofold dilutions of a reference serum to test doubling dilutions of antigen preparations is recommended for standardizing different lots of antigen. Using the above-mentioned standardized agarose plates and appropriate reference serum with anti-Sm and/or anti-RNP antibodies in a chessboard titration, it is recommended that acceptable use dilution of antigen preparation be two doubling dilutions before reaching an endpoint. The reference sera selected should be compared to US reference standards supplied by the Center for Disease Control.

Antibodies, Antinuclear↗
PubMed1987

Evidence against ras activation in human ovarian carcinomas.

Activation of ras genes in human ovarian carcinomas and ovarian carcinoma cell lines was tested by transfection of NIH 3T3 cells with high molecular weight DNA extracted from fresh tumors or from cell lines. Of 18 ovarian tumors and tumor cell lines that were exhaustively tested, only one yielded DNA active in focus-induction in this assay. Southern blot analysis of DNA in serially transformed NIH 3T3 foci revealed restriction fragments that hybridized with a probe specific for the human N-ras oncogene and with human repetitive (Alu) sequences. Individually transformed foci contained N-ras hybridizing fragments of different sizes. Northern blots of RNA extracted from different transformants revealed the expression of different transcripts hybridizing with the N-ras probe. The transformants also expressed a 700 base RNA that hybridized with a human B-lym probe. The low prevalence of ras activation in ovarian tumors as measured by transforming capability suggests that ras activation does not play an important role in the development of most human ovarian tumors.

Animals↗
PubMed1998

A method for examining differential mRNA expression along the crypt-villus axis of the human small intestine.

1. The aim of this study was to devise a method of segregating crypt and villus cell subpopulations from endoscopic human small intestinal biopsies which might be used to examine changes associated with functional differentiation at the molecular level. 2. Routine endoscopic biopsies from the human small intestine were subjected to a modified protocol of mechanical disruption and chelation to yield subpopulations of different cell types. The purity and character of the cell populations isolated was assessed by measuring enzyme activity and thymidine incorporation and by histology. A guanidinium isothiocyanate method was adapted for small samples to extract RNA from the isolated subpopulations, and probes for RNA with a known predilection for crypt and villus cells were used to further investigate the application and usefulness of the technique. 3. Sequential histological examination during the segregation protocol demonstrated that different cell types were removed serially from the biopsy samples. Cell-type enrichment of the segregated subpopulations was demonstrated by differential alkaline phosphatase activity and by differences in thymidine incorporation in the samples isolated. Sufficient quantities of RNA could be extracted from the segregated subpopulations for Northern blot analysis and the differential expression of mRNA for sucrase-isomaltase and transferrin receptor was demonstrated in the villus and crypt subpopulations respectively. 4. Messenger RNA can be successfully extracted from different cell types segregated from routine human endoscopic small intestinal biopsies. This technique should prove useful for investigating the mechanisms regulating the functional differentiation of epithelial cells in the small intestine and the regulatory mechanisms governing absorption of macromolecules.

Biopsy↗
PubMed1989

[A case of a so-called congenital tooth appeared in maxilla].

The congenital teeth are not only classified by Massler et al. as a natal tooth that has erupted at birth and a neonatal tooth that erupts at neonatal period, but also divided into a normal deciduous tooth and a supernumerary tooth. The congenital teeth are mostly found in the mandibular anterior region, but extremely rare in other regions. The authors met a rare case, that had a so-called congenital tooth in the maxillary anterior region, which was a supernumerary and fused tooth. The case was a boy patient, 4 months of age, who visited the pedodontic clinic, of Ohu University Dental Hospital, with the complaint of ruber and swelling of the maxillary anterior gum pad, October 17th in 1986. By oral inspection of the dental radiogram and his past history, the authors found a odontoid material in the swelled gum pad and diagnosed it as a so-called congenital and supernumerary tooth. The tooth was extracted. The extracted tooth was investigated histopathologically. It had the v-shaped incisula at the middle of its edge, and seemed as if two teeth had coalesced. The serial preparations by the labio-lingual section showed no partition, such cementum as to divide the enamel, the dentin and the pulp near the incisula. Therefore it was diagnosed as fused tooth correctly. In the labial surface of the root formed about a third root length, the thickness of the cellular cementum was observed. It was considered as a physiological reaction to no support of the alveolar bone and to the excess mobility of the tooth. The prognosis after its extraction was good, and all normal deciduous incisors appeared in his mouth, 7 month later.

Fused Teeth↗
PubMed1972

Axenic culture of myxamoebae of the myxomycete Physarum polycephalum.

Myxamoebae of the acellular slime mold Physarum polycephalum have been cultured axenically in a soluble medium. The growth medium contains bovine serum albumin, embryo extracts, liver infusion broth, peptone, and glucose. Cell densities ranging from 3 x 10(6) to 5 x 10(6) cells/ml have been obtained with this medium. To date, myxamoebae have been serially transferred more than 100 times without deleterious effect.

Agar↗
PubMed1988

Serial oxytocin levels in amniotic fluid and maternal plasma during normal and induced labour.

Oxytocin was measured by a specific and sensitive radioimmunoassay in plasma and amniotic fluid after extraction with Sep-Pak cartridges in patients undergoing elective caesarean sections, normal labour and labour induced with oxytocin infusion or prostaglandins. In maternal plasma, levels of oxytocin correlated with the period of gestation; concentrations at term were significantly higher than in the first two trimesters. Maternal plasma levels of oxytocin before the onset of contractions were not significantly different from those at the onset of spontaneous labour or at full cervical dilatation. Levels of oxytocin in patients induced with oxytocin were not statistically different from levels observed in spontaneous labour. Amniotic fluid oxytocin levels were not significantly different between the groups either at the onset of labour or immediately before delivery. Umbilical arterio-venous differences in oxytocin were present in all groups except the prostaglandin-induced group. Increased prostaglandins associated with the onset of normal labour are probably not a consequence of changes in oxytocin concentrations.

Amniotic Fluid↗