Genetic factors influencing the risk of multiple myeloma bone disease.

A major complication of multiple myeloma (MM) is the development of osteolytic lesions, fractures and bone pain. To identify genetic variants influencing the development of MM bone disease (MBD), we analyzed MM patients of European ancestry (totaling 3774), which had been radiologically surveyed for MBD. Each patient had been genotyped for ~6 00 000 single-nucleotide polymorphisms with genotypes for six million common variants imputed using 1000 Genomes Project and UK10K as reference. We identified a locus at 8q24.12 for MBD (rs4407910, OPG/TNFRSF11B, odds ratio=1.38, P=4.09 × 10(-9)) and a promising association at 19q13.43 (rs74676832, odds ratio=1.97, P=9.33 × 10(-7)). Our findings demonstrate that germline variation influences MBD and highlights the importance of RANK/RANKL/OPG pathway in MBD development. These findings will contribute to the development of future strategies for prevention of MBD in the early precancerous phases of MM.

Preparation and antiarthritic and analgesic activity of 4,5-diaryl-2-(substituted thio)-1H-imidazoles and their sulfoxides and sulfones.

A series of 4,5-diaryl-2-(substituted thio)-1H-imidazoles was synthesized and evaluated as antiinflammatory and analgesic agents in the rat adjuvant induced arthritis assay and the mouse phenyl-p-benzoquinone writhing (PQW) assay. Several analogues were found to be more potent than phenylbutazone and indomethacin. Structure-activity relationships are discussed. One of the compounds, 4,5-bis(4-fluorophenyl)-2-[(1,1,2,2-tetrafluoroethyl)-sulfonyl]-1H- imidazole (8d, tiflamizole), was found to be 8 times as potent as indomethacin in the rat adjuvant induced arthritis assay and is presently undergoing clinical trial as an antiarthritic drug.

Antibacterials. Synthesis and structure-activity studies of 3-aryl-2-oxooxazolidines. 1. The "B" group.

The synthesis and structure/activity studies of the effect of varying the "B" group in a series of oxazolidinone antibacterials (I) are described. Two synthetic routes were used: (1) alkylation of aniline with glycidol followed by dialkyl carbonate heterocyclization to afford I (A = H, B = OH), whose arene ring was further elaborated by using electrophilic aromatic substitution methodology; (2) cycloaddition of substituted aryl isocyanates with epoxides to give A and B with a variety of values. I with B = OH or Br were converted to other "B" functionalities by using SN2 methodology. Antibacterial evaluation of compounds I with A = acetyl, isopropyl, methylthio, methylsulfinyl, methylsulfonyl, and sulfonamido and a variety of different "B" groups against Staphylococcus aureus and Enterococcus faecalis concluded that the compounds with B = aminoacyl, and particularly acetamido, were the most active of those examined in each A series, possessing MICs in the range of 0.5-4 micrograms/mL for the most active compounds described.

Antibacterials. synthesis and structure-activity studies of 3-aryl-2-oxooxazolidines. 4. Multiply-substituted aryl derivatives.

The synthesis and structure-activity relationship (SAR) studies of the effect of different polysubstitution patterns in the aromatic ring of 5-(acetamidomethyl)oxazolidinone antibacterials (I) on antibacterial activity are presented. Compounds I were prepared by the six-step synthesis described previously (Gregory, W. A.; et al. J. Med. Chem. [formula: see text] 1989, 32, 1673), electrophilic aromatic substitution reactions of 3-substituted compounds, and functional-group interchange reactions of 3,4-disubstituted compounds. Antibacterial evaluation of compounds I against Staphylococcus aureus and Enterococcus faecalis gave the following results. The 2,4- and 2,5-disubstituted derivatives have weak or no antibacterial activity. Antibacterial activities of 3,4-disubstituted compounds are comparable to those of the 4-monosubstituted analogues for small 3-substituents (smaller than Br), but decline rapidly for larger 3-substituents. 3,4-Annulated derivatives are comparable in activity to their open-chain analogues. 3,5-Disubstituted and 3,4,5- and 2,4,6-trisubstituted derivatives are devoid of antibacterial activity.

Antibacterials. Synthesis and structure-activity studies of 3-aryl-2-oxooxazolidines. 2. The "A" group.

The synthesis and structure-activity relationship (SAR) studies of the effect of varying the "A" group in a series of 5-(acetamidomethyl)oxazolidonone antibacterials (2) are described. Compounds 2 were principally prepared either by the six-step synthesis described previously (J. Med. Chem. 1989, 32, 1673) or by elaboration of the synthetic intermediate 2 (A = H) via electrophilic aromatic substitution or elaboration of the intermediate 2 (A = I) by transition metal catalyzed carbon-carbon bond-forming reactions. Antibacterial evaluation of compounds 2 with A = alkyl, ethenyl, ethynyl, hydroxyalkyl, aldo and keto, oximinoakyl, carboalkoxy, nitro, amino, halo and psi-halo, alkylthio, alkylsulfinyl, and alkysulfonyl against Staphylococcus aureus and Enterococcus faecalis led to the identification of several SAR trends. In several series of homologues (alkyl, ketyl, aximinoalkyl, amino, halo and psi-halo, and alkythio), antibacterial activity increased with increasing lipophilicity. But in series with where A is a substituent with a trior tetrasubstituted (substituent larger than H) quaternary atom attached directly to the aromatic ring (hydroxyalkyl, alkylsulfinyl, alkylsufonyl), the activity peaked at the member of the series with the "tert-butyl" connectivity pattern. Conjugated electron-withdrawing substituents also had increased activity relative to nonconjugated analogues of comparable lipophilicity.

Maturation of the goldfish preoptic area: ultrastructural correlates of position and maturation of neurosecretory cells.

A gradation of small rostral to larger caudal neurosecretory cells is found in the preoptic area of fish. Regional variations in ultrastructural features were assessed in this region of fish ranging widely in size (1.7-283.1 g), some of which had received intraperitoneal injections of horseradish peroxidase in order to label neurons projecting beyond the blood-brain barrier (i.e. neurosecretory cells). In small (less than or equal to 6.6 g) fish, neurosecretory cells of the caudal preoptic area were 10-15 microns in diameter and contained dense core vesicles. More rostral smaller cells had few or no dense core vesicles, but were labeled following intraperitoneal horseradish peroxidase injections. Cells in both areas were progressively larger, with larger nuclei and more granular reticulum in larger fish. In large fish, neurons that contained dense core vesicles ranged from 10 microns in diameter rostrally to 70 microns in diameter caudally. Some intermediate and large cells were extensively vacuolated and contained extremely convoluted and possibly multiple nuclei. Degenerating material was seen in apparently normal large fish. A few neurosecretory cells were extensively surrounded by perineuronal electron-dense glial cells. Most neurosecretory cells were involved in extensive soma-somatic apposition, which was especially pronounced in fish which were removed from cold water in late fall. Gap junctions were also clearly present on the somata of preoptic neurosecretory cells of these fall fish. These data imply that increased capacity for hormone secretion in large fish may be accomplished in part by neuronal hypertrophy. Maturational and seasonal variation in structure suggest that physiological characteristics are also variable in this nucleus.

Horseradish peroxidase evidence for a spinal projection from the preoptic area of the goldfish, a light and electron microscopic study.

Horseradish peroxidase (HRP) was applied to the transected spinal cord of goldfish labeled neurons in the preoptic area. Since leakage of HRP into the blood could produce the labeling of neurosecretory cells, intraperitoneal (i.p.) injections of HRP were made with a wide range of dosages in order to intentionally label preoptic neurosecretory cells. The distribution of preoptic neurons labeled after spinal HRP application was far more restricted than the labeling via uptake of HRP from the blood, even when cells in the spinal cord-transected fish were intensely labeled. Furthermore, in HRP electron microscopic material, morphological differences were observed between neurons labeled by the two procedures. Large numbers of dense core vesicles and well-developed stacks of rough endoplasmic reticulum, features typical of cells projecting to the pituitary, were not observed in cells labeled via the spinal cord. These findings indicate that in goldfish a direct projection exists from the preoptic area to the spinal cord which could be homologous to one arising from the paraventricular nucleus of mammals. Both i.p. injection and spinal transection also produced labeling of more caudal periventricular diencephalic cells which resemble preoptic cells in efferent projections as well as ultrastructural features.

Gap junctions in goldfish preoptic ependyma: regional variation in cellular differentiation.

Ependyma adjacent to the goldfish preoptic neurosecretory nucleus was examined with transmission electron microscopy. Ependymal cells adjoining the rostroventral end of the nucleus were spindle-shaped with their long axes perpendicular to the ventricular surface. Gap junctions and desmosomes were common near the apical (ventricular) ends of these cells, and less frequent laterally in the ependymal layer. Ependymal cells in more caudodorsal preoptic regions (adjacent to large neurosecretory cells) were progressively more pleomorphic. The frequent occurrence of apparently internalized gap junctions and of gap junction fragments enclosed within lysosome-like organelles indicated extensive turnover of these junctions, or uncoupling. Ependymal cells in the caudodorsal region formed gap junctions on their lateral and basal (abluminal) surfaces with glial processes containing bundles of intermediate filaments. Subependymally, these processes (presumptive radial glia) were parallel to one another and coupled together by gap junctions. Neurites containing dense core vesicles occasionally invaginated into ependymal cells in the caudal region, but did not appear to form gap junctions. Previous observations indicate continuing maturation and growth of the goldfish preoptic area with neurosecretory cell formation rostroventrally and a rostroventral to caudodorsal gradient of maturation. The present findings suggest a parallel and related gradient in preoptic ependyma. Ependymal cell differentiation possibly involves loss of gap junctions, and radial migration or differentiation into underlying neurons and glia.

Satellite glial cells penetrate neurosecretory cells to perinuclear position in the goldfish preoptic area.

Some goldfish neurosecretory cells have plasma membrane invaginations filled by processes of surrounding satellite glial cells (SCs) that produce trophospongium-like multicellular neuron-glial aggregates. Some penetrating SC processes approach the neuronal nucleus, reaching to within approximately 40 nm of the outer nuclear membrane. Gap junctions were found in one freeze-fracture replica through an apparent neuronal-glial aggregate, suggesting that neuron-glial gap junctions may be present. The extensive covering and penetration of these neurons by SCs suggests trophic relationships and communication by undetermined modalities between neurons and glia. The common close proximity of invaginated SC processes to the neuronal nucleus may indicate that information is transmitted between SCs and the nucleus. Some SCs abut against the basal lamina of large blood vessels and contain dense vesicles, either secretory or lysosomal.

Neuronal and glial gap junctions in the goldfish preoptic area, a thin section and freeze-fracture study.

In freeze-fracture, both large macular gap junctions and long thin gap junctions surrounded by a strand of tight junction were found on neurosecretory cells. Preoptic neurons show large areas of soma-to-soma apposition, but thin section showed no evidence for gap junctions between neuronal somata. Neurosecretory cell neurites formed parallel bundles in neuropil lateral to the nucleus, and gap junctions were found between the neurites. These junctions apparently correspond to macular junctions seen on neurosecretory elements in freeze-fracture. Some large macular gap junctions found in freeze-fracture presumably correspond to junctions seen between glial cells in thin section. However, glial membranes lacked characteristics distinguishing them from neuronal membranes. In one instance, a large apparent glial sheet process formed both macular and long thin gap junctions on different surfaces. The long thin gap junctions that were surrounded by a strand of tight junction were formed with a large neurosecretory cell soma. Extensive pinocytosis was observed at some membranes forming gap junctions.

Ultrastructure of neurons in the paraventricular nucleus of normal, dehydrated and rehydrated rats.

Previous reports have shown that the percentage of neuronal somatic membrane in soma-somatic apposition (without intervening glia) increased with brief periods of dehydration (4--24 hr) and decreased with rehydration in the rat supraoptic and circularis nuclei. In the present study, the percentage of somal membrane in soma-somatic appositions was found to increase in the primarily vasopressin-containing lateral portion of the rat paraventricular nucleus with twelve hours of dehydration. Further evidence for altered cellular function in this nucleus was a decrease in the number of smaller dense core vesicles (< 2600A) per unit cytoplasmic area during initial dehydration (4--12 hr). No changes were detected, however, in the number of larger dense core vesicles (> 4000 A) or lysosomes (> 4000 A) per unit cytoplasm. Intranuclear membrane-bound vacuoles were found primarily in hydrated and rehydrated animals. No reliable changes were seen in the dilation of granular endoplasmic reticulum. Cilia were found in the neuropil and were occasionally traced to magnocellular somata. Differences in the patterns of morphological responses among the magnocellular hypothalamic nuclei suggest specializations in their roles, and further support a functional significance of neuronal membrane appositions.

Spontaneous and osmotically-stimulated activity in slices of rat hypothalamus.

Single unit activity was recorded from 400-500 mu m thick slices of rat hypothalamus, using either NaCl- or horseradish peroxidase-filled glass micropipettes. Spontaneous activity was present in the following hypothalamic loci: anterior hypothalamic-preoptic area, nucleus circularis, nucleus of the diagonal band of Broca, paraventricular accessory nucleus, paraventricular nucleus (all portions), periventricular regions of the anterior hypothalamus, and the suprachiasmatic nucleus. The supraoptic nucleus was the only major cell group studied to exhibit no spontaneous activity. Cells of the paraventricular and circularis nuclei were spontaneously active, displayed firing rates and patterns of activity similar to those recorded in vivo for magnocellular elements of the hypothalamus, and in some cases responded to increases in the osmolality of the bathing medium with altered firing rates and/or patterns of activity. Many cells in these preparations were characterized by phasic, bursting patterns of activity. Slow, irregular and regular, continuous activity was also frequently observed, as is typical in vivo. Median firing rates were in the range of 4-6 spikes/sec, somewhat faster than the rates usually reported for anesthetized in vivo preparations. These rates are more similar to those observed in unanesthetized monkeys or rats with diencephalic islands. Extracellular HRP marking provided a high degree of localization for many of the recorded cells. These results indicate that the hypothalamic slice preparation is useful for studies in which it is desirable to eliminate extrahypothalamic connections and in which it is necessary to exercise a fine degree of control over the extracellular environment of the cells.

Nucleolar proliferation and cell size changes in rat supraoptic neurons following osmotic and volemic challenges.

Subcutaneous injections of isotonic saline induced nucleolar proliferation in supraoptic neurons in animals sacrificed approximately 5 min postinjection. The magnitude of this proliferation was sustained 4 and 8 hr postinjection. Polyethylene glycol (PG) injections depleted blood volume 4 and 8 hr after the injection, but the percentage of SON cells with multiple nucleoli in these animals was not different from saline-injected controls. The anterior (SOa) portion of the SON in rats given 2% NaCl to drink instead of water for three days contained more cells with multiple nucleoli than controls. This effect was enhanced after five days ingestion, and accompanied by a similar response in the tuberal portion of SON (SOt). Rehydration for ten days after three days of 2% NaCl intake brought the percentage of cells with multiple nucleoli down to control levels. Cell area in SON cells paralleled nucleolar responses during dehydration and rehydration. The results demonstrate the sensitivity of nucleolar proliferation in SON to environmental changes ranging from osmotic to neurogenic stress.

Effects of heterogeneous layers of composite and time on composite repair of porcelain.

Bond strengths of porcelain/composite resin repair samples, some homogeneous of conventional, hybrid, and microfill materials, some heterogeneous, made by incremental build-up of two of these composites, were evaluated. Samples were allowed to set without disturbance. After storage in 37 degrees C water for intervals of one day, seven days, and 28 days, the test samples were subjected to tensile force until fracture. There were significant differences in bond strengths of homogenous and heterogeneous samples after different storage periods. All mean repair bond strengths were significantly less at seven days than at one or 28 days. Heterogeneous repairs with larger-particle-size composite at the porcelain interface and overlayed with smaller-particle-size composites resulted in higher bond strengths than the homogeneous small-sized composite repairs. Failures of the repairs occurred at the porcelain/composite interface in a statistically significant number, implying that technique exactness at the interface plays an important role in the success of the technique.

Quantitative evaluation of approximal contacts in Class 2 composite resin restorations: a clinical study.

The purpose of this study was to develop an accurate and reproducible intraoral method of measuring the distance between two teeth, and to conduct a clinical pilot study of the effect of placement of a class 2 posterior resin restoration upon this intertooth dimension. A Kaman Sciences KD-2611 noncontact displacement measuring system with a 1U unshielding sensor, based upon the variable resistance of eddy current, was used for the intraoral measurement. Seven patients requiring conservative interproximal restorations (initial placements or replacements) on the premolars were selected. Addition silicone impressions were taken of the posterior quadrants from which low-fusing metal (Cerrolow-136) casts were made. Composite resin copings constructed on modified rubber dam clamps were fabricated for the restoration and adjacent teeth upon these metal casts. Aluminum targets and Plexiglas holders for the sensor were attached to the bow of the rubber dam clamp with light-activated composite resin. Quantitative measurements of intertooth distance were made preoperatively, postoperatively, and at one-week and 4 1/2-month recalls after placement of class 2 composite resin restorations for 11 teeth. Four unrestored premolars served as a control. The data were tabulated and analyzed using one-way analysis of variance. The results indicated that the mean postoperative intercoping distance for restored teeth increased after a multiple wedging technique had been applied during restorative procedure. At the one-week recall, these distances had decreased. The 4 1/2-month measurements showed a further decrease with an average loss of 15 microns from the preoperative baseline.

Interim luting agents, composite core surface hardness and retention of interim and final restorations.

This study compared the effects of ZOE interim cement on the retention of interim acrylic and final cast crowns (cemented with zinc phosphate) over cores of current conventional and hybrid composite resin formulae, and a non-eugenol interim cement (NOG) and a calcium hydroxide liner (CaOH2) on interim and final retention values over hybrid cores. Results were compared to retention of cast crowns over amalgam cores. Retention was measured with an Instron Testing Machine. Location of cement failure during separation was recorded. Surface hardness of each group was compared. Retention of cast crowns cemented with zinc phosphate over amalgam cores was significantly higher than that of any of the resin core groups (ANOVA, P less than 0.05, Sheffe Contrast). There was no significant difference in final casting retention cemented over hybrid or conventional resin cores exposed to ZOE (13.625 vs 14.125 Kg). Final casting retention differences were not significant between the three interim cement groups. Final retention of all groups exposed to an interim luting agent were significantly less than the composite control groups. The use of CaOH, as an interim luting agent for acrylic crowns over hybrid cores compared to ZOE or NOG, should afford significantly greater retention with no adverse effect on the retention of the final casting. While the surface hardness of both types of composite resin was adversely affected by exposure to interim luting agents as compared to controls, there was no correlation between final casting retention values and surface hardness of polished composite. The interim cements invariably failed at the interim acrylic crown interface while the zinc phosphate cement failed at the core interface.

Effects of intra-core mechanical interlocks and cement type on full crown retention.

This study compared the retention of full cast crowns over cores of amalgam or a Bis-GMA composite resin when cemented with a Bis-GMA composite resin cement or a zinc phosphate cement. The surfaces of two groups of Bis-GMA composite cores were mechanically altered with a groove or dimples to provide physical retention; two other groups were either etched with 1.23% acidulated phosphate fluoride (APF) or exposed to a zinc oxide-eugenol interim luting agent before crown cementation. Control groups of unaltered composite or amalgam core served as comparison standards. The zinc phosphate cement provided significantly greater retention with the amalgam core than with any of the composite cores. The APF-treated composite cores had the least retention of any of the zinc phosphate cementations. Cementation failure over the composite cores was at the core surface. Crown retention over the control and dimpled-surface composite core with the resin cement was twice that with zinc phosphate, and significantly greater than retention of castings cemented with the resin cement over amalgam cores. Retention over amalgam cores was significantly greater with zinc phosphate cement than with the composite resin cement.

Effect of light source position and bevel placement on facial margin adaptation of resin-based composite restorations.

PURPOSE: To evaluate the effect of direction of first cure and the presence or absence of a facial margin bevel on the adaptation of facial margins of Class III resin-based composite (RBC) restorations. MATERIALS AND METHODS: Custom polyvinyl siloxane (PVS) light shields were fabricated for both the mesial and distal surfaces of 20 extracted incisors. Class III cavities were prepared in the mesial and distal surfaces of the incisors. On the facial, lingual, and gingival margins of 20 of the preparations a 0.5 mm wide enamel bevel was placed. The lingual and gingival, but not the facial, margins were beveled on the other 20 preparations. The PVS light shield was adapted to the tooth and trimmed to allow 1-2 mm of the facial margin to be exposed on 10 of the beveled and 10 of the non-beveled preparations. For the remaining 20 preparations the PVS light shield was trimmed to allow access of the curing light only from the lingual. Teeth were restored with Prisma TPH RBC. Using the appropriate shield, restorations in the facial first cure group were cured first from the facial and then from the lingual. Restorations in the lingual first cure group were cured only from the lingual. Restorations were finished flush with enamel margins and thermocycled following storage. Samples were exposed to a 50% solution of silver nitrate followed by light exposure. Teeth were sectioned and microleakage was evaluated at three levels (incisal, middle, and cervical). Four evaluators (blinded to direction of cure) independently scored microleakage using a categorical evaluation scale. Statistical analysis included non-parametric descriptive statistics, Cohen's kappa, chi-square analysis, and Kruskal-Wallis ANOVA. RESULTS: Samples prepared without a facial bevel revealed significantly more (P = 0.01) microleakage than preparations with a facial bevel. At the most incisal level, restorations cured first from the facial revealed significantly more (P < 0.001) microleakage than restorations cured first from the lingual. At the middle and cervical levels, restorations cured first from the facial revealed significantly less (P < 0.001) microleakage than restorations cured first from the lingual. When a facial bevel was present, samples cured first from the facial showed significantly less (P < 0.001) microleakage than those cured only from the lingual.